Home Medicine MARK1 suppress malignant progression of hepatocellular carcinoma and improves sorafenib resistance through negatively regulating POTEE
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MARK1 suppress malignant progression of hepatocellular carcinoma and improves sorafenib resistance through negatively regulating POTEE

  • Xin Lu , Zhiyuan Chen , Wenting Mi , Jianming Zheng and Yubin Liu EMAIL logo
Published/Copyright: November 7, 2024

Abstract

Purpose

This study aimed to investigate the role of microtubule-affinity regulatory protein kinase 1 (MARK1) in hepatocellular carcinoma (HCC) progression, its association with sorafenib sensitivity, and the interplay between MARK1 and POTE Ankyrin domain family member E(POTEE) in HCC cells.

Methods

Quantitative real-time polymerase chain reaction analysis was used to assess MARK1 and POTEE expression in 60 pairs of HCC tissues and cell lines. The correlation between MARK1 levels, clinicopathological features, and patient prognosis was analyzed. Sorafenib-resistant HCC cell models were developed, followed by MARK1 overexpression to evaluate its impact on cell functions. Luciferase reporter assays and rescue experiments were conducted to elucidate the MARK1–POTEE regulatory mechanism.

Results

MARK1 exhibited decreased mRNA expression in HCC tissues and cells, correlating with adverse clinicopathological features and poorer patient survival. Luciferase assays confirmed direct binding between MARK1 and POTEE. Sorafenib treatment increased MARK1 protein levels, reduced POTEE, and inhibited cell proliferation. Overexpressing MARK1 suppressed sorafenib-induced proliferation in resistant cells, while co-overexpression of MARK1 and POTEE reversed this effect.

Conclusion

MARK1 potentially restrains HCC progression and enhances sorafenib resistance by negatively modulating POTEE expression, highlighting its significance as a therapeutic target in HCC treatment.

1 Introduction

Hepatocellular carcinoma (HCC) is the second leading cause of death induced by cancer in the world, with its characteristics including insidious onset, high recurrence rate, and strong metastasis [1,2]. It is the third largest malignancy in China, which accounts for more than half of the global cases [3,4]. Therefore, it is of great clinical significance to establish a reasonable treatment scheme for HCC and explore new treatment strategies [5,6]. Hepatitis B and C virus infections are leading causes of HCC, particularly in Asia and Africa. Chronic infection with these viruses leads to liver inflammation, cirrhosis, and eventually HCC. Given the high global burden of these infections, it is critical to consider them in HCC pathogenesis [7]. The immune system also plays a critical role in the development and progression of HCC. Pro-inflammatory cytokines, such as TNF-α and IL-6, promote carcinogenesis by activating various signaling pathways. These pathways contribute to tumor cell proliferation, survival, and invasion. Many studies have highlighted the importance of inflammation in cancer development, which we discuss in further detail in this study [8,9]. At present, the precise treatment of tumors, especially molecular targeted drug treatments, has attracted more and more attention. Its unique anti-tumor effect is gradually revealed, and its application in liver cancer has also achieved initial results [10,11]. However, unlike other tumors, there are large heterogeneities in molecular signaling pathways and biological characteristics among different subpopulations of liver cancer, for example, BRAF gene mutations in melanoma account for nearly half, which directly leads to limitation of molecular targeted drugs [12,13].

Existing clinical evidence have showed that sorafenib, as a leader of molecular-targeted drugs, could remarkably extend the survival time of patients with advanced liver cancer and has been first approved by the FDA of USA for the treatment of advanced liver cancer [14,15]. However, due to its limited sensitivity and effectiveness and the great individual differences, its efficacy is far from meeting patients’ expectations [15,16]. Sorafenib is only sensitive to 30% of patients with liver cancer, while the rest 70% patients show different degrees of resistance to sorafenib treatment. Although a number of clinical trials of sorafenib combined therapy have been carried out, no better targeted drug has been found to treat sorafenib-resistant liver cancer [17,18]. Therefore, the identification of biological targets is related to sorafenib sensitivity and individualized treatment of patients with different liver cancer subgroups. On the one hand, it greatly saves social medical resources, and on the other hand, it avoids unnecessary overtreatment of patients [18,19]. In addition, exploring the role of specific molecular targets in the sorafenib sensitivity of HCC cells and the underlying mechanisms can help to provide new ideas for individualized treatment of HCC [19,20].

Microtubule-affinity regulatory protein kinase (MARK), a novel mammalian serine/threonine kinase, is a microtubule-related protein binding domain kinase [21,22]. It is involved in the phosphorylation of tau, MAP2, and MAP4 proteins in microtubule-related fields, leading to microtubule-decomposition and increasing dynamic changes [22]. We explored the role of MARK1 in the malignant progression of HCC and its effect on HCC cell resistance to sorafenib, and further discussed the mutual regulation mechanism between MARK1 and POTEE.

2 Patients and methods

2.1 Patients and HCC samples

Tumor tissue specimens and adjacent ones of 60 HCC patients undergoing surgical resection were collected. All subjects had not received any radiotherapy or chemotherapy before surgery. HCC classification was based on the 8th edition UICC/AJCC liver cancer TNM staging criteria, which was used to categorize the patients according to tumor size, nodal involvement, and distant metastasis [23].

2.2 Cell culture

Human-derived HCC cells (MHCC97H, SMMC-7221, Huh7, Hep3B) and a human normal liver cell line (HL-7702) provided by American Type Culture Collection (Manassas, VA, USA) were cultured with Dulbecco’s Modified Eagle’s Medium (Gibco, Rockville, MD, USA) supplemented with 10% fetal bovine serum (Gibco, Rockville, MD, USA), streptomycin (100 μg/mL), and penicillin (100 U/mL) in an incubator with 5% CO2 at 37°C.

2.3 Transfection

Lipofectamine 3000 reagent (Invitrogen, Carlsbad, CA, USA) was mixed with MARK1 overexpression sequence (MARK1) (GenePharma, Shanghai, China) and then added into cells when cell density reached 70%. After 48 h, cells were collected for subsequent analysis.

2.4 Cell counting kit-8 (CCK-8) assay

After 48 h of transfection, cells were seeded into 60-well plates (2,000 cells/well), and CCK-8 test (Dojindo Molecular Technologies, Kumamoto, Japan) was carried out to detect cell proliferation capacity based on the instructions.

2.5 Colony formation experiment

After transfection for 48 h, 200 cells were seeded in each well of a 6-well plate and cultured with complete medium for 2 weeks. After that, the cells were cloned and fixed in 2 mL of methanol for 20 min. After the methanol was aspirated, the cells were stained with 0.1% crystal violet, photographed, and counted under a light-selective environment.

2.6 Quantitative real-time polymerase chain reaction (qPCR)

qPCR detection was implemented based on the instructions of SYBR® Premix Ex Taq ™ on StepOne Plus Real-time PCR System (TaKaRa, Tokyo, Japan), with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as internal reference. Primers used in the qPCR reaction were as follows: MARK1: forward: ′-AGCCAGGCAGTGATTTGAGG-3′, reverse: 5′-CAGTAACGAGGGAGGGCTTC-3′; POTEE: forward: 5′-GTACCACGTCCGTGGAGAAG-3′, reverse: 5′-TGTAGAGCAGTCCTCTTTTGC-3′; GAPDH: forward: 5′-CCTGGCACCCAGCACAAT-3′, reverse: 5′-GCTGATCCACATCTGCTGGAA-3′.

2.7 Western blot

Western blot analysis was performed according to standard procedures. The primary antibodies against MARK1, POTEE, and GAPDH, and the secondary antibodies anti-mouse and anti-rabbit were all purchased from Cell Signaling Technology (Danvers, MA, USA). Protein expression levels were quantified using densitometry analysis, and the results were presented as graphs in the corresponding figures. We used ImageJ software to analyze the protein bands, following the protocol outlined in previous work [24].

2.8 Dual-luciferase reporter assay

HCC cell lines Huh7 and Hep3B were co-transfected with POTEE/NC and pMIR luciferase reporter plasmids using Lipofectamine 2000. After 48 h of transfection, the dual luciferase reporter assay system (Promega, Madison, WI, USA) was used to normalize the reporter luciferase activity to control firefly luciferase activity.

2.9 Statistical analysis

GraphPad Prism 5 V5.01 software (La Jolla, CA, USA) was used for statistical analysis. Differences between two groups were analyzed by using Student’s t-test. Comparison between multiple groups was done using one-way ANOVA test followed by post-hoc test (least significant difference). Each experiment is repeated at least for three independent experiments, and the data are expressed as X ± SD. P < 0.05 was considered statistically significant.

  1. Ethical approval: This study was approved by the ethics committee of Guangdong Provincial People’s Hospital.

  2. Informed consent: Signed written informed consents were obtained from the patients and/or guardians.

3 Results

3.1 Expression of MARK1 and POTEE in HCC

To determine the role of MARK1 and POTEE in HCC, a total of 60 pairs of tumor and para-cancer tissue samples from HCC patients were collected and detected their expression in these samples. We found a reduction in MARK1 expression and an increase in POTEE expression in HCC tissues in comparison to the adjacent ones (Figure 1a and c), suggesting that MARK1 may serve as a tumor suppressor gene in this cancer. At the same time, the same alternation in the above two gene levels was observed in HCC cell lines (Figure 1b and d).

Figure 1 
                  MARK1 and POTEE expression in HCC tissues and cell lines. (a) qRT-PCR was used to detect the difference in the expression of MARK1 in tumor tissues and non-tumor tissues adjacent to the cancer of HCC patients. (b) qRT-PCR was used to detect the expression level of MARK1 in HCC cell lines. (c) qRT-PCR was used to detect the difference in the expression of POTEE in tumor tissues and non-tumor tissues adjacent to cancer of HCC patients. (d) qRT-PCR was used to detect the expression level of POTEE in HCC cell lines. Data are average ± SD, *P < 0.05, **P < 0.01, ***P < 0.001.
Figure 1

MARK1 and POTEE expression in HCC tissues and cell lines. (a) qRT-PCR was used to detect the difference in the expression of MARK1 in tumor tissues and non-tumor tissues adjacent to the cancer of HCC patients. (b) qRT-PCR was used to detect the expression level of MARK1 in HCC cell lines. (c) qRT-PCR was used to detect the difference in the expression of POTEE in tumor tissues and non-tumor tissues adjacent to cancer of HCC patients. (d) qRT-PCR was used to detect the expression level of POTEE in HCC cell lines. Data are average ± SD, *P < 0.05, **P < 0.01, ***P < 0.001.

3.2 MARK1 expression is closely related to the pathological stage of HCC patients

HCC tissues collected from 60 HCC patients were divided into high MARK1 expression and low expression group, and further explored the relationship between MARK1 expression and clinicopathological parameters of HCC patients by chi-square test. Table 1 shows a positive correlation between lowly expressed MARK1 and the pathological stage of HCC, but not with age, gender, incidence of lymph node, or distant metastasis.

Table 1

Association of MARK1 expression with clinicopathological characteristics of HCC

Parameters Number of cases MARK1 expression p-Value
High (n = 36) Low (n = 24)
Age (years) 0.389
<60 24 16 8
≥60 36 20 16
Gender 0.058
Male 29 21 8
Female 31 15 16
T stage 0.008
T1–T2 35 26 9
T3–T4 25 10 15
Lymph node metastasis 0.914
No 37 22 15
Yes 23 14 9
Distance metastasis 0.598
No 48 28 20
Yes 12 8 4

3.3 POTEE is the target gene of MARK1

Bioinformatics analysis suggested that POTEE is a potential target gene of MARK1, which was further verified by the results obtained from luciferase reporter gene experiments (Figure 2a). In addition, we demonstrated a negative correlation between the mRNA level of MARK1 and POTEE (Figure 2b).

Figure 2 
                  MARK1 directly targets POTEE. (a) The dual luciferase reporter gene experiment verified the direct targeting of MARK1 to POTEE. (b) Expression levels of MARK1 and POTEE in HCC tissues were significantly negatively correlated. Data are average ± SD, **P < 0.01.
Figure 2

MARK1 directly targets POTEE. (a) The dual luciferase reporter gene experiment verified the direct targeting of MARK1 to POTEE. (b) Expression levels of MARK1 and POTEE in HCC tissues were significantly negatively correlated. Data are average ± SD, **P < 0.01.

3.4 Sorafenib inhibits the growth of HCC cells

We then examined the sorafenib sensitivity of HCC cells in Huh7 and Hep3B cell lines by CCK-8 test. As a result, sorafenib dose-dependently suppressed the proliferative ability of HCC cells (Figure 3a). In addition, western blot showed that MARK1 protein level was significantly increased while that of POTEE protein was oppositely reduced in sorafenib-resistant HCC cells compared to normal HCC cells (Figure 3b).

Figure 3 
                  MARK1 can increase the sorafenib sensitivity of HCC cell lines. (a) CCK-8 was used to evaluate the cell proliferation of HCC cell lines Huh7 and Hep3B after treatment of different concentrations of sorafenib. (b) Western blot detected the expression levels of MARK1 and POTEE in normal HCC cells and sorafenib-resistant HCC cells. Data are average ± SD, *P < 0.05.
Figure 3

MARK1 can increase the sorafenib sensitivity of HCC cell lines. (a) CCK-8 was used to evaluate the cell proliferation of HCC cell lines Huh7 and Hep3B after treatment of different concentrations of sorafenib. (b) Western blot detected the expression levels of MARK1 and POTEE in normal HCC cells and sorafenib-resistant HCC cells. Data are average ± SD, *P < 0.05.

3.5 Overexpression of MARK1 suppresses the proliferation of sorafenib-resistant HCC cells

Administration of vectors to overexpress MARK1 successfully increases the protein level of MARK1 in sorafenib-resistant cell lines (Figure 4a). Figure 4b shows that overexpressing MARK1 markedly attenuated the proliferation ability of the sorafenib-resistant cells. And the plate cloning experiment also revealed a consistent reduced cell proliferation to CCK8 results (Figure 4c).

Figure 4 
                  Proliferation of sorafenib-resistant liver cancer cells was inhibited after overexpression of MARK1. (a) Western blot verified the transfection efficiency of sorafenib-resistant HCC cell lines Huh7 and Hep3B after transfection of MARK1 overexpression vector. (b) CCK-8 test was used to detect the cell proliferation ability after transfection of MARK1 overexpression vector in sorafenib-resistant HCC cell lines Huh7 and Hep3B. (c) Plate cloning experiment detected the number of HCC positive proliferating cells (magnification: 40×) after transfection of MARK1 overexpression vector in sorafenib-resistant HCC cell lines Huh7 and Hep3B. Data are average ± SD, *P < 0.05.
Figure 4

Proliferation of sorafenib-resistant liver cancer cells was inhibited after overexpression of MARK1. (a) Western blot verified the transfection efficiency of sorafenib-resistant HCC cell lines Huh7 and Hep3B after transfection of MARK1 overexpression vector. (b) CCK-8 test was used to detect the cell proliferation ability after transfection of MARK1 overexpression vector in sorafenib-resistant HCC cell lines Huh7 and Hep3B. (c) Plate cloning experiment detected the number of HCC positive proliferating cells (magnification: 40×) after transfection of MARK1 overexpression vector in sorafenib-resistant HCC cell lines Huh7 and Hep3B. Data are average ± SD, *P < 0.05.

3.6 POTEE can reverse influence of MARK1 on the proliferative ability of sorafenib-resistant HCC cells

To further test the interaction between MARK1 and POTEE in sorafenib-resistant cell lines, we used a combination of plasmids to overexpress MARK1 and POTEE in vitro. Western blot results confirmed that MARK1 level was remarkably reduced after co-transfection of MARK1 and POTEE overexpression vectors (Figure 5a). Subsequently, both the CCK-8 and the plate cloning experiments showed that overexpression of POTEE could counteract the inhibitory impact of overexpression of MARK1 on sorafenib-resistant HCC cell proliferation capacity (Figure 5b and c).

Figure 5 
                  MARK1 can regulate the expression of POTEE in sorafenib-resistant HCC cells. (a) Western blot detected the expression of MARK1 after co-transfection of MARK1 and POTEE overexpression vectors in sorafenib-resistant HCC cell lines Huh7 and Hep3B. (b) CCK-8 experiment was used to detect the effect of co-transfection of MARK1 and POTEE overexpression vectors on proliferation of sorafenib-resistant HCC cells. (c) Plate cloning experiment detected the number of HCC positive proliferating cells (magnification: 40×) after co-transfection of MARK1 and POTEE overexpression vectors in sorafenib-resistant HCC cell lines Huh7 and Hep3B. Data are average ± SD, *P < 0.05.
Figure 5

MARK1 can regulate the expression of POTEE in sorafenib-resistant HCC cells. (a) Western blot detected the expression of MARK1 after co-transfection of MARK1 and POTEE overexpression vectors in sorafenib-resistant HCC cell lines Huh7 and Hep3B. (b) CCK-8 experiment was used to detect the effect of co-transfection of MARK1 and POTEE overexpression vectors on proliferation of sorafenib-resistant HCC cells. (c) Plate cloning experiment detected the number of HCC positive proliferating cells (magnification: 40×) after co-transfection of MARK1 and POTEE overexpression vectors in sorafenib-resistant HCC cell lines Huh7 and Hep3B. Data are average ± SD, *P < 0.05.

4 Discussion

HCC is one of the most common malignancies in China and even the world, ranking fifth in the incidence of global tumors and third in mortality [1,2,3]. Although great progress has been made in the treatment of HCC with chemotherapy drugs in recent years, the clinical application of chemotherapy drugs has been seriously hindered due to the huge difference in efficacy among different HCC patients, which has become a major obstacle to the treatment of HCC [4,5,6]. Sorafenib, an oral multi-kinase inhibitor, is a targeted drug used in the clinical treatment of advanced HCC in recent years. Tumor angiogenesis is blocked by inhibiting vascular endothelial growth factor receptor and platelet-derived growth factor receptor [15,16,17,18]. Although sorafenib is able to remarkably extend the survival time of some HCC patients, with the emergence of primary and acquired resistance to sorafenib, the exploration of biological targets related to sorafenib sensitivity in HCC becomes increasingly important [18,19,20].

In this study, a large number of clinical samples of HCC patients for the first time were used to investigate the role of MARK1 and POTEE in the development of this cancer. qPCR results showed that MARK1 expression in HCC cancer tissues and cell lines was decreased compared to that in para-carcinoma tissues and normal liver cell lines, while POTEE was increased to varying degrees, indicating that MARK1 and POTEE play an essential part in the progression of HCC. To further verify the relationship between MARK1 and sorafenib sensitivity of HCC cells, we conducted CCK-8 test, which then demonstrated that sorafenib suppresses the proliferative capacity. Meanwhile, MARK1 level in HCC cells with sorafenib resistance was remarkably increased, while POTEE level was decreased. To verify the influence of MARK1 on the biological behavior of sorafenib-resistant HCC cell lines, we overexpressed MARK1 and measured the proliferation of these cells. As a result, we found that MARK1 is able to inhibit the proliferation capacity of HCC and plays a pivotal role in HCC, but the specific molecular mechanism remains unclear. Luciferase reporter assay verified that MARK1 can directly bind to its target gene POTEE, and we demonstrated that overexpression of POTEE could reverse the suppressing influence of overexpression of MARK on the proliferative ability of sorafenib-resistant HCC cells. Therefore, this study suggests that MARK1 may inhibit the malignant progression of HCC and promote sorafenib resistance through regulating POTEE.

This study is just a preliminary study on part of the mechanism of MARK1’s negative regulation of POTEE on liver cancer cells. The specific mechanism is much more complex than we imagined, and further research is needed to provide new ideas for the treatment of liver cancer.

5 Conclusions

This study suggests that MARK1 may inhibit malignant progression of HCC and increase HCC cell resistance to sorafenib through negatively modulating POTEE expression.


# Xin Lu, Zhiyuan Chen and Wenting Mi contributed equally to this work.

tel: +86 013826288388

  1. Funding information: This study was funded by the Guangdong Natural Science Foundation (2019A15155011733), which was received by Yubin Liu and supported by grants from the Guangzhou Science and Technology Plan Project (2023A04J2385), which was received by Wenting Mi.

  2. Author contributions: Xin Lu, Zhiyuan Chen, and Wenting Mi: conceptualization, methodology, writing ‒ original draft preparation; Jianming Zheng: investigation, software, statistical analysis; Yubin Liu: reviewing and editing, funding acquisition, supervision. All authors read and approved the final manuscript.

  3. Conflict of interest: The authors have no conflicts with any step of the article preparation.

  4. Data availability statement: The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Received: 2024-06-11
Revised: 2024-09-07
Accepted: 2024-09-23
Published Online: 2024-11-07

© 2024 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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  57. Clinical characteristics and assessment of risk factors in patients with influenza A-induced severe pneumonia after the prevalence of SARS-CoV-2
  58. Analgesia nociception index is an indicator of laparoscopic trocar insertion-induced transient nociceptive stimuli
  59. High STAT4 expression correlates with poor prognosis in acute myeloid leukemia and facilitates disease progression by upregulating VEGFA expression
  60. Factors influencing cardiovascular system-related post-COVID-19 sequelae: A single-center cohort study
  61. HOXD10 regulates intestinal permeability and inhibits inflammation of dextran sulfate sodium-induced ulcerative colitis through the inactivation of the Rho/ROCK/MMPs axis
  62. Mesenchymal stem cell-derived exosomal miR-26a induces ferroptosis, suppresses hepatic stellate cell activation, and ameliorates liver fibrosis by modulating SLC7A11
  63. Endovascular thrombectomy versus intravenous thrombolysis for primary distal, medium vessel occlusion in acute ischemic stroke
  64. ANO6 (TMEM16F) inhibits gastrointestinal stromal tumor growth and induces ferroptosis
  65. Prognostic value of EIF5A2 in solid tumors: A meta-analysis and bioinformatics analysis
  66. The role of enhanced expression of Cx43 in patients with ulcerative colitis
  67. Choosing a COVID-19 vaccination site might be driven by anxiety and body vigilance
  68. Role of ICAM-1 in triple-negative breast cancer
  69. Cost-effectiveness of ambroxol in the treatment of Gaucher disease type 2
  70. HLA-DRB5 promotes immune thrombocytopenia via activating CD8+ T cells
  71. Efficacy and factors of myofascial release therapy combined with electrical and magnetic stimulation in the treatment of chronic pelvic pain syndrome
  72. Efficacy of tacrolimus monotherapy in primary membranous nephropathy
  73. Mechanisms of Tripterygium wilfordii Hook F on treating rheumatoid arthritis explored by network pharmacology analysis and molecular docking
  74. FBXO45 levels regulated ferroptosis renal tubular epithelial cells in a model of diabetic nephropathy by PLK1
  75. Optimizing anesthesia strategies to NSCLC patients in VATS procedures: Insights from drug requirements and patient recovery patterns
  76. Alpha-lipoic acid upregulates the PPARγ/NRF2/GPX4 signal pathway to inhibit ferroptosis in the pathogenesis of unexplained recurrent pregnancy loss
  77. Correlation between fat-soluble vitamin levels and inflammatory factors in paediatric community-acquired pneumonia: A prospective study
  78. CD1d affects the proliferation, migration, and apoptosis of human papillary thyroid carcinoma TPC-1 cells via regulating MAPK/NF-κB signaling pathway
  79. miR-let-7a inhibits sympathetic nerve remodeling after myocardial infarction by downregulating the expression of nerve growth factor
  80. Immune response analysis of solid organ transplantation recipients inoculated with inactivated COVID-19 vaccine: A retrospective analysis
  81. The H2Valdien derivatives regulate the epithelial–mesenchymal transition of hepatoma carcinoma cells through the Hedgehog signaling pathway
  82. Clinical efficacy of dexamethasone combined with isoniazid in the treatment of tuberculous meningitis and its effect on peripheral blood T cell subsets
  83. Comparison of short-segment and long-segment fixation in treatment of degenerative scoliosis and analysis of factors associated with adjacent spondylolisthesis
  84. Lycopene inhibits pyroptosis of endothelial progenitor cells induced by ox-LDL through the AMPK/mTOR/NLRP3 pathway
  85. Methylation regulation for FUNDC1 stability in childhood leukemia was up-regulated and facilitates metastasis and reduces ferroptosis of leukemia through mitochondrial damage by FBXL2
  86. Correlation of single-fiber electromyography studies and functional status in patients with amyotrophic lateral sclerosis
  87. Risk factors of postoperative airway obstruction complications in children with oral floor mass
  88. Expression levels and clinical significance of serum miR-19a/CCL20 in patients with acute cerebral infarction
  89. Physical activity and mental health trends in Korean adolescents: Analyzing the impact of the COVID-19 pandemic from 2018 to 2022
  90. Evaluating anemia in HIV-infected patients using chest CT
  91. Ponticulus posticus and skeletal malocclusion: A pilot study in a Southern Italian pre-orthodontic court
  92. Causal association of circulating immune cells and lymphoma: A Mendelian randomization study
  93. Assessment of the renal function and fibrosis indexes of conventional western medicine with Chinese medicine for dredging collaterals on treating renal fibrosis: A systematic review and meta-analysis
  94. Comprehensive landscape of integrator complex subunits and their association with prognosis and tumor microenvironment in gastric cancer
  95. New target-HMGCR inhibitors for the treatment of primary sclerosing cholangitis: A drug Mendelian randomization study
  96. Population pharmacokinetics of meropenem in critically ill patients
  97. Comparison of the ability of newly inflammatory markers to predict complicated appendicitis
  98. Comparative morphology of the cruciate ligaments: A radiological study
  99. Immune landscape of hepatocellular carcinoma: The central role of TP53-inducible glycolysis and apoptosis regulator
  100. Serum SIRT3 levels in epilepsy patients and its association with clinical outcomes and severity: A prospective observational study
  101. SHP-1 mediates cigarette smoke extract-induced epithelial–mesenchymal transformation and inflammation in 16HBE cells
  102. Acute hyper-hypoxia accelerates the development of depression in mice via the IL-6/PGC1α/MFN2 signaling pathway
  103. The GJB3 correlates with the prognosis, immune cell infiltration, and therapeutic responses in lung adenocarcinoma
  104. Physical fitness and blood parameters outcomes of breast cancer survivor in a low-intensity circuit resistance exercise program
  105. Exploring anesthetic-induced gene expression changes and immune cell dynamics in atrial tissue post-coronary artery bypass graft surgery
  106. Empagliflozin improves aortic injury in obese mice by regulating fatty acid metabolism
  107. Analysis of the risk factors of the radiation-induced encephalopathy in nasopharyngeal carcinoma: A retrospective cohort study
  108. Reproductive outcomes in women with BRCA 1/2 germline mutations: A retrospective observational study and literature review
  109. Evaluation of upper airway ultrasonographic measurements in predicting difficult intubation: A cross-section of the Turkish population
  110. Prognostic and diagnostic value of circulating IGFBP2 in pancreatic cancer
  111. Postural stability after operative reconstruction of the AFTL in chronic ankle instability comparing three different surgical techniques
  112. Research trends related to emergence agitation in the post-anaesthesia care unit from 2001 to 2023: A bibliometric analysis
  113. Frequency and clinicopathological correlation of gastrointestinal polyps: A six-year single center experience
  114. ACSL4 mediates inflammatory bowel disease and contributes to LPS-induced intestinal epithelial cell dysfunction by activating ferroptosis and inflammation
  115. Affibody-based molecular probe 99mTc-(HE)3ZHER2:V2 for non-invasive HER2 detection in ovarian and breast cancer xenografts
  116. Effectiveness of nutritional support for clinical outcomes in gastric cancer patients: A meta-analysis of randomized controlled trials
  117. The relationship between IFN-γ, IL-10, IL-6 cytokines, and severity of the condition with serum zinc and Fe in children infected with Mycoplasma pneumoniae
  118. Paraquat disrupts the blood–brain barrier by increasing IL-6 expression and oxidative stress through the activation of PI3K/AKT signaling pathway
  119. Sleep quality associate with the increased prevalence of cognitive impairment in coronary artery disease patients: A retrospective case–control study
  120. Dioscin protects against chronic prostatitis through the TLR4/NF-κB pathway
  121. Association of polymorphisms in FBN1, MYH11, and TGF-β signaling-related genes with susceptibility of sporadic thoracic aortic aneurysm and dissection in the Zhejiang Han population
  122. Application value of multi-parameter magnetic resonance image-transrectal ultrasound cognitive fusion in prostate biopsy
  123. Laboratory variables‐based artificial neural network models for predicting fatty liver disease: A retrospective study
  124. Decreased BIRC5-206 promotes epithelial–mesenchymal transition in nasopharyngeal carcinoma through sponging miR-145-5p
  125. Sepsis induces the cardiomyocyte apoptosis and cardiac dysfunction through activation of YAP1/Serpine1/caspase-3 pathway
  126. Assessment of iron metabolism and iron deficiency in incident patients on incident continuous ambulatory peritoneal dialysis
  127. Tibial periosteum flap combined with autologous bone grafting in the treatment of Gustilo-IIIB/IIIC open tibial fractures
  128. The application of intravenous general anesthesia under nasopharyngeal airway assisted ventilation undergoing ureteroscopic holmium laser lithotripsy: A prospective, single-center, controlled trial
  129. Long intergenic noncoding RNA for IGF2BP2 stability suppresses gastric cancer cell apoptosis by inhibiting the maturation of microRNA-34a
  130. Role of FOXM1 and AURKB in regulating keratinocyte function in psoriasis
  131. Parental control attitudes over their pre-school children’s diet
  132. The role of auto-HSCT in extranodal natural killer/T cell lymphoma
  133. Significance of negative cervical cytology and positive HPV in the diagnosis of cervical lesions by colposcopy
  134. Echinacoside inhibits PASMCs calcium overload to prevent hypoxic pulmonary artery remodeling by regulating TRPC1/4/6 and calmodulin
  135. ADAR1 plays a protective role in proximal tubular cells under high glucose conditions by attenuating the PI3K/AKT/mTOR signaling pathway
  136. The risk of cancer among insulin glargine users in Lithuania: A retrospective population-based study
  137. The unusual location of primary hydatid cyst: A case series study
  138. Intraoperative changes in electrophysiological monitoring can be used to predict clinical outcomes in patients with spinal cavernous malformation
  139. Obesity and risk of placenta accreta spectrum: A meta-analysis
  140. Shikonin alleviates asthma phenotypes in mice via an airway epithelial STAT3-dependent mechanism
  141. NSUN6 and HTR7 disturbed the stability of carotid atherosclerotic plaques by regulating the immune responses of macrophages
  142. The effect of COVID-19 lockdown on admission rates in Maternity Hospital
  143. Temporal muscle thickness is not a prognostic predictor in patients with high-grade glioma, an experience at two centers in China
  144. Luteolin alleviates cerebral ischemia/reperfusion injury by regulating cell pyroptosis
  145. Therapeutic role of respiratory exercise in patients with tuberculous pleurisy
  146. Effects of CFTR-ENaC on spinal cord edema after spinal cord injury
  147. Irisin-regulated lncRNAs and their potential regulatory functions in chondrogenic differentiation of human mesenchymal stem cells
  148. DMD mutations in pediatric patients with phenotypes of Duchenne/Becker muscular dystrophy
  149. Combination of C-reactive protein and fibrinogen-to-albumin ratio as a novel predictor of all-cause mortality in heart failure patients
  150. Significant role and the underly mechanism of cullin-1 in chronic obstructive pulmonary disease
  151. Ferroptosis-related prognostic model of mantle cell lymphoma
  152. Observation of choking reaction and other related indexes in elderly painless fiberoptic bronchoscopy with transnasal high-flow humidification oxygen therapy
  153. A bibliometric analysis of Prader-Willi syndrome from 2002 to 2022
  154. The causal effects of childhood sunburn occasions on melanoma: A univariable and multivariable Mendelian randomization study
  155. Oxidative stress regulates glycogen synthase kinase-3 in lymphocytes of diabetes mellitus patients complicated with cerebral infarction
  156. Role of COX6C and NDUFB3 in septic shock and stroke
  157. Trends in disease burden of type 2 diabetes, stroke, and hypertensive heart disease attributable to high BMI in China: 1990–2019
  158. Purinergic P2X7 receptor mediates hyperoxia-induced injury in pulmonary microvascular endothelial cells via NLRP3-mediated pyroptotic pathway
  159. Investigating the role of oviductal mucosa–endometrial co-culture in modulating factors relevant to embryo implantation
  160. Analgesic effect of external oblique intercostal block in laparoscopic cholecystectomy: A retrospective study
  161. Elevated serum miR-142-5p correlates with ischemic lesions and both NSE and S100β in ischemic stroke patients
  162. Correlation between the mechanism of arteriopathy in IgA nephropathy and blood stasis syndrome: A cohort study
  163. Risk factors for progressive kyphosis after percutaneous kyphoplasty in osteoporotic vertebral compression fracture
  164. Predictive role of neuron-specific enolase and S100-β in early neurological deterioration and unfavorable prognosis in patients with ischemic stroke
  165. The potential risk factors of postoperative cognitive dysfunction for endovascular therapy in acute ischemic stroke with general anesthesia
  166. Fluoxetine inhibited RANKL-induced osteoclastic differentiation in vitro
  167. Detection of serum FOXM1 and IGF2 in patients with ARDS and their correlation with disease and prognosis
  168. Rhein promotes skin wound healing by activating the PI3K/AKT signaling pathway
  169. Differences in mortality risk by levels of physical activity among persons with disabilities in South Korea
  170. Review Articles
  171. Cutaneous signs of selected cardiovascular disorders: A narrative review
  172. XRCC1 and hOGG1 polymorphisms and endometrial carcinoma: A meta-analysis
  173. A narrative review on adverse drug reactions of COVID-19 treatments on the kidney
  174. Emerging role and function of SPDL1 in human health and diseases
  175. Adverse reactions of piperacillin: A literature review of case reports
  176. Molecular mechanism and intervention measures of microvascular complications in diabetes
  177. Regulation of mesenchymal stem cell differentiation by autophagy
  178. Molecular landscape of borderline ovarian tumours: A systematic review
  179. Advances in synthetic lethality modalities for glioblastoma multiforme
  180. Investigating hormesis, aging, and neurodegeneration: From bench to clinics
  181. Frankincense: A neuronutrient to approach Parkinson’s disease treatment
  182. Sox9: A potential regulator of cancer stem cells in osteosarcoma
  183. Early detection of cardiovascular risk markers through non-invasive ultrasound methodologies in periodontitis patients
  184. Advanced neuroimaging and criminal interrogation in lie detection
  185. Maternal factors for neural tube defects in offspring: An umbrella review
  186. The chemoprotective hormetic effects of rosmarinic acid
  187. CBD’s potential impact on Parkinson’s disease: An updated overview
  188. Progress in cytokine research for ARDS: A comprehensive review
  189. Utilizing reactive oxygen species-scavenging nanoparticles for targeting oxidative stress in the treatment of ischemic stroke: A review
  190. NRXN1-related disorders, attempt to better define clinical assessment
  191. Lidocaine infusion for the treatment of complex regional pain syndrome: Case series and literature review
  192. Trends and future directions of autophagy in osteosarcoma: A bibliometric analysis
  193. Iron in ventricular remodeling and aneurysms post-myocardial infarction
  194. Case Reports
  195. Sirolimus potentiated angioedema: A case report and review of the literature
  196. Identification of mixed anaerobic infections after inguinal hernia repair based on metagenomic next-generation sequencing: A case report
  197. Successful treatment with bortezomib in combination with dexamethasone in a middle-aged male with idiopathic multicentric Castleman’s disease: A case report
  198. Complete heart block associated with hepatitis A infection in a female child with fatal outcome
  199. Elevation of D-dimer in eosinophilic gastrointestinal diseases in the absence of venous thrombosis: A case series and literature review
  200. Four years of natural progressive course: A rare case report of juvenile Xp11.2 translocations renal cell carcinoma with TFE3 gene fusion
  201. Advancing prenatal diagnosis: Echocardiographic detection of Scimitar syndrome in China – A case series
  202. Outcomes and complications of hemodialysis in patients with renal cancer following bilateral nephrectomy
  203. Anti-HMGCR myopathy mimicking facioscapulohumeral muscular dystrophy
  204. Recurrent opportunistic infections in a HIV-negative patient with combined C6 and NFKB1 mutations: A case report, pedigree analysis, and literature review
  205. Letter to the Editor
  206. Letter to the Editor: Total parenteral nutrition-induced Wernicke’s encephalopathy after oncologic gastrointestinal surgery
  207. Erratum
  208. Erratum to “Bladder-embedded ectopic intrauterine device with calculus”
  209. Retraction
  210. Retraction of “XRCC1 and hOGG1 polymorphisms and endometrial carcinoma: A meta-analysis”
  211. Corrigendum
  212. Corrigendum to “Investigating hormesis, aging, and neurodegeneration: From bench to clinics”
  213. Corrigendum to “Frankincense: A neuronutrient to approach Parkinson’s disease treatment”
  214. Special Issue The evolving saga of RNAs from bench to bedside - Part II
  215. Machine-learning-based prediction of a diagnostic model using autophagy-related genes based on RNA sequencing for patients with papillary thyroid carcinoma
  216. Unlocking the future of hepatocellular carcinoma treatment: A comprehensive analysis of disulfidptosis-related lncRNAs for prognosis and drug screening
  217. Elevated mRNA level indicates FSIP1 promotes EMT and gastric cancer progression by regulating fibroblasts in tumor microenvironment
  218. Special Issue Advancements in oncology: bridging clinical and experimental research - Part I
  219. Ultrasound-guided transperineal vs transrectal prostate biopsy: A meta-analysis of diagnostic accuracy and complication rates
  220. Assessment of diagnostic value of unilateral systematic biopsy combined with targeted biopsy in detecting clinically significant prostate cancer
  221. SENP7 inhibits glioblastoma metastasis and invasion by dissociating SUMO2/3 binding to specific target proteins
  222. MARK1 suppress malignant progression of hepatocellular carcinoma and improves sorafenib resistance through negatively regulating POTEE
  223. Analysis of postoperative complications in bladder cancer patients
  224. Carboplatin combined with arsenic trioxide versus carboplatin combined with docetaxel treatment for LACC: A randomized, open-label, phase II clinical study
  225. Special Issue Exploring the biological mechanism of human diseases based on MultiOmics Technology - Part I
  226. Comprehensive pan-cancer investigation of carnosine dipeptidase 1 and its prospective prognostic significance in hepatocellular carcinoma
  227. Identification of signatures associated with microsatellite instability and immune characteristics to predict the prognostic risk of colon cancer
  228. Single-cell analysis identified key macrophage subpopulations associated with atherosclerosis
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