1.The effect of TNF-α-regulated circWBSCR22 on the migration and invasion of cervical cancer cells through the miR-512-5p/FAM60A axis
Lili BAI ; Jin ZHANG ; Yatong FAN ; Pengyu ZHANG
Acta Universitatis Medicinalis Anhui 2026;61(4):689-698
ObjectiveTo explore the effect and mechanism of tumor necrosis factor alpha (TNF-α)-regulated circular RNA Williams Beuren syndrome chromosomal region 22 (circWBSCR22) on the malignant behaviors of cervical cancer cells. MethodsHuman cervical cancer HeLa cells were treated with TNF-α. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression level of circWBSCR22. Nuclear-cytoplasmic RNA separation experiments were conducted to determine the localization of circWBSCR22 in cells. Cell migration, invasion, viability, and proliferation were assessed through Transwell assay, tetrazolium salt (MTT) assay, and colony formation assay, respectively. Dual-luciferase reporter gene assays were performed to verify the targeting relationship between microRNA-512-5p (miR-512-5p) and circWBSCR22 or family with sequence similarity 60A (FAM60A). RT-qPCR was used to detect the RNA levels of miR-512-5p and FAM60A. Western blot was used to detect the protein level of FAM60A. ResultsCompared with the Control group, the expression level of circWBSCR22 in HeLa cells treated with TNF-α increased (P<0.05). About 66% of circWBSCR22 was found in the cytoplasm. Compared with the control group, the migration (P<0.01) and invasion (P<0.01) of HeLa cells transfected with circWBSCR22 were enhanced, but the cell activity and cell proliferation were not significantly affected (P>0.05). The double luciferase reporter gene assay showed that the relative luciferase activity of the co transfected circWBSCR22-WT and miR-512-5p groups was lower than that of the co transfected circWBSCR22-WT and miR-NC groups (P<0.01). Compared with the control group, the expression level of miR-512-5p in HeLa cells transfected with circWBSCR22 decreased (P<0.01). Compared with the control group, the migration (P<0.01) and invasion (P<0.05) ability of HeLa cells transfected with miR-512-5p decreased. Double luciferase reporter gene assay showed that compared with the co transfected FAM60A 3´-UTR-WT and miR-NC groups, the fluorescence activity of the co transfected FAM60A 3´-UTR-WT and miR-512-5p groups decreased (P<0.01).Compared with the control group, the expression levels of FAM60A mRNA (P<0.05) and protein (P<0.01) in HeLa cells transfected with miR-512-5p decreased. Compared with the control group, the migration (P<0.01) and invasion (P<0.01) of cervical cancer HeLa cells transfected with FAM60A were enhanced. Compared with the control group, the expression levels of FAM60A mRNA (P<0.01) and protein (P<0.05) in the cells transfected with circWBSCR22 increased. The results of rescue experiment showed that compared with the pcDNA+miR-NC group, the expression levels of FAM60A RNA (P<0.01) and protein (P<0.001) in the pcDNA+miR-512-5p group decreased, and the ability of migration (P<0.01) and invasion (P<0.05) decreased; Compared with pcDNA+miR-512-5p group, the expression levels of FAM60A RNA (P<0.01) and protein (P<0.001) in circWBSCR22+miR-512-5p group increased, and the ability of migration (P<0.01) and invasion (P<0.05) was enhanced. ConclusionTNF-α promotes the migration and invasion of cervical cancer cells through the circWBSCR22/miR-512-5p/FAM60A regulatory axis.
2.DIA Proteomic Profiling on Staged Regulatory Effect of Tonifying Deficiency and Dredging Collaterals Method on Liver Fibrosis in Rats Based on Theory of "Zhu Ke Jiao"
Xin WANG ; Pengyu ZHU ; Li WEN ; Jibin LIU ; Aochun YUE ; Ziyi CHEN ; Jing ZHANG ; Li ZHU ; Quansheng FENG ; Cen JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):119-132
ObjectiveThis paper aims to investigate the differential mechanisms underlying the staged therapeutic effects of Qijia Rougan formula on liver fibrosis using proteomic technology. MethodsThe staged rat model of liver fibrosis was established by subcutaneous injection of carbon tetrachloride (CCl4) and olive oil. One hundred and four SD rats were randomized into thirteen groups:a normal group,a two-week model group,a four-week model group,a six-week model group,an eight-week model group,a two-week Qijia Rougan formula group,a four-week Qijia Rougan formula group,a six-week Qijia Rougan formula group,an eight-week Qijia Rougan formula group,a two-week compound Biejia Ruangan tablet group,a four-week Compound Biejia Ruangan Tablet group,a six-week Compound Biejia Ruangan Tablet group,and an eight-week compound Biejia Ruangan tablet group. After two weeks of drug intervention,liver tissue and abdominal aortic blood samples were collected from the rats for testing. Hematoxylin-eosin (HE) staining,Masson staining,and Picro Sirius red staining were used to observe pathological damage and collagen fiber deposition in liver tissues. Immunohistochemistry (IHC) was employed to detect the contents of fibrosis markers in liver tissues. The contents of liver function indicators in the serum were measured using a fully automated biochemical analyzer,and the levels of liver fibrosis indicators in the serum were assessed by enzyme-linked immunosorbent assay (ELISA). Liver tissues from the normal group,each model group,and each Qijia Rougan formula group were subjected to label-free quantitative proteomic analysis to identify differential proteins among the groups,with key proteins validated by Western blot. Finally,bioinformatics analysis was performed on the differential proteins. Results(1) The staged rat model of liver fibrosis constructed with CCl4 and olive oil showed pathological results at the 2nd,4th,6th,and 8th weeks of modeling that were consistent with the Metavir standards for the F1,F2,F3,and F4 stages. Compared with those in the normal control group,the protein expressions of α-smooth muscle actin (α-SMA) and Collagen Ⅰ were significantly increased in each stage (P<0.05). The levels of liver function indicators in the serum,including alanine aminotransferase (ALT),aspartate aminotransferase (AST),alkaline phosphatase (ALP),direct bilirubin (DBIL),and total bilirubin (TBil) in each model group,were significantly elevated in each stage (P<0.01). The levels of liver fibrosis indicators in the serum,including procollagen Ⅲ peptide (PⅢP),type Ⅳ collagen(Ⅳ-C),hyaluronic acid (HA),and laminin (LN) in each model group,were significantly increased in each stage (P<0.05,P<0.01). This study successfully established a staged rat model of liver fibrosis. (2) Compared with the model groups at each stage,the administration groups showed a reduction in hepatocyte ballooning degeneration,a more orderly arrangement of hepatocytes,and a decrease of inflammatory cell infiltration. The blue-stained collagen fibers became significantly thinner and finer,with reduced and narrowed fibrous septa. The areas of collagen fibers and Picro Sirius red staining were reduced (P<0.05). The positive areas of α-SMA and Collagen Ⅰ expression were significantly decreased (P<0.05). The levels of ALT,AST,ALP,DBIL,and TBil in the rats of the model groups at each stage were significantly reduced (P<0.05,P<0.01). The levels of PⅢP,Ⅳ-C,HA,and LN in the rats of the model groups at each stage were significantly decreased (P<0.05). Among these,the improvements in all indicators were most significant in the F3 stage (P<0.01).(3) The proteomic results show that a total of 165 differential proteins exhibit a callback trend when comparing the model groups at four stages with the normal group,and when comparing the Qijia Rougan formula group with the model group. Western blot analysis reveals that the levels of NAD(P)H:quinone oxidoreductase 1 (NQO1),mitogen-activated protein kinase 1 (MAPK1),arginase 1 (Arg1),and glutathione S-transferase α1 (GSTA1) were consistent with the proteomic results. Bioinformatics results reveal that 165 differentially expressed proteins are enriched in multiple signaling pathways. Notably,signaling pathways such as drug metabolism-cytochrome P450,arginine biosynthesis,and the peroxisome proliferator-activated receptor (PPAR) signaling pathway were found to be closely associated with liver fibrosis,suggesting that the Qijia Rougan formula may exert its staged regulatory effects on liver fibrosis by regulating these pathways. ConclusionThe Qijia Rougan formula may achieve staged regulation of liver fibrosis by regulating drug metabolism-cytochrome P450,arginine biosynthesis,and the PPAR signaling pathway.
3.Expert consensus on electronic patient-reported outcome-based symptom management for perioperative lung cancer patients (version 2026)
Wei DAI ; Cheng LEI ; Yuanqiang ZHANG ; Rong ZHANG ; Pengyu Jinming ; Jinming XU ; Yuzhen ZHENG ; Liang ZHAO ; Guibin QIAO ; Guowei CHE ; Jian HU ; Lei JIANG ; Jie LI ; Qiang LI ; Qiuling SHI
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(08):1166-1178
Patients with lung cancer experience a heavy symptom burden during the perioperative period, which seriously affects their recovery and quality of life. Traditional symptom management models rely mainly on scheduled ward rounds during hospitalization and outpatient follow-up after discharge. However, they have limitations such as delayed symptom recognition, lack of post-discharge monitoring, and non-quantitative symptom assessment, which often lead to delayed interventions and low patient satisfaction. In recent years, the symptom management model based on electronic patient-reported outcomes (ePRO) has been increasingly valued in clinical practice. Existing high-level evidence from both domestic and international studies indicates that, through proactive monitoring, real-time alerts, and remote interventions, this model enables dynamic and continuous symptom management and helps improve patient recovery and healthcare experience. As a supplement to routine medical care, the ePRO-based symptom management model aims to enhance the quality of care rather than replace existing medical processes. To promote the standardized application of this model in perioperative lung cancer care, this consensus integrates domestic and international evidence. After multiple rounds of voting by more than 50 experts, it formulates 12 consensus statements covering the three core components, symptom monitoring, alerting, and intervention, to provide scientific and practical recommendations for clinical practice.
4.Current status and prospect of precision treatment for colorectal cancer
Hongwei YAO ; Jiale GAO ; Zhengyang YANG ; Liting SUN ; Pengyu WEI ; Zhongtao ZHANG
Chinese Journal of Digestive Surgery 2025;24(6):690-694
In recent years, with continuous advancements in molecular biology and gene testing technologies, the diagnosis and treatment of colorectal cancer have been rapidly transitioning toward precision medicine. The application of molecular classification, target detection, and liquid biopsy technologies has driven ongoing updates to clinical guidelines. Multidisciplinary team colla-boration, innovations in precision surgical techniques, and the widespread adoption of neoadjuvant combination therapies have collectively promoted more individualized and scientific management of colorectal cancer. Looking ahead,the authors believe that as multi-omics biomarkers, organoid models, and artificial intelligence are increasingly integrated into clinical practice, precision diagnosis and treatment of colorectal cancer will deepen further, offering patients more efficient and personalized therapeutic options.
5.Clinical characteristics of locally advanced rectal cancer patients with pathological complete response after neoadjuvant chemoradiotherapy combined with immunotherapy: a national multicenter study
Jiale GAO ; Yuanyuan2 YANG ; Zhengyang YANG ; Jiagang3 HAN ; Ang? LI ; Gang? LIU ; Yi? SUN ; Liting SUN ; Pengyu WEI ; Jianyong ZHENG ; Hongwei YAO ; Zhongtao ZHANG
Chinese Journal of Digestive Surgery 2025;24(6):739-745
Objective:To analyze the clinical characteristics of locally advanced rectal cancer patients with pathological complete response (pCR) after neoadjuvant chemoradiotherapy combined with immunotherapy.Methods:The retrospective cohort study was conducted. The clinicopatholo-gical data of 46 patients with locally advanced rectal cancer who were admitted to 6 medical centers, including Beijing Friendship Hospital of Capital Medical University et al, from June 2021 to November 2022 were collected. There were 29 males and 17 females, aged (61±4)years. Patients received neoadjuvant chemoradiotherapy combined with immune checkpoint inhibitor therapy, and under-went radical total mesorectal excision during 6-12 weeks after radiotherapy. Observation indicators: (1) comparison of clinical characteristics between pCR and non-pCR patients;(2) postoperative complications and adverse reactions of pCR and non-pCR patients. Comparison of measurement data with normal distribution between groups was conducted using the t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data between groups was conducted using the Mann-Whitney U test. Results:(1) Comparison of clinical characteristics between pCR and non-pCR patients. Before neoadjuvant therapy, there were 14 cases aged ≥50 years and 6 cases aged <50 years in pCR patients, versus 25 cases and 1 case in non-pCR patients, showing a significant difference between the two groups ( P<0.05). After neoadjuvant therapy, cases in clinical stage T0, T1, T2, T3, T4 were 11, 1, 5, 3, 0 for pCR patients versus 7, 4, 2, 11, 2 for non-pCR patients, cases of tumor regression grade 1, 2, 3, 4 were 11, 8, 1, 0 for pCR patients versus 7, 14, 4, 1 for non-pCR patients, cases in low-risk, medium-risk, high-risk of neoadjuvant rectal scoring and grading were 20, 0, 0 for pCR patients versus 4, 18, 4 for non-pCR patients, respectively, showing significant differences in above indicators between the two groups ( Z=-2.256, -2.104, -5.458, P<0.05). (2) Postoperative complications and adverse reactions of pCR and non-pCR patients. Postoperative complications occurred in 2 cases of pCR patients and 5 cases of non-pCR patients, postoperative adverse reactions occurred in 11 cases of pCR patients and 10 cases of non-pCR patients, showing no significant difference between the two groups ( P>0.05). Conclusion:Compared with locally advanced rectal cancer patients aged ≥50 years, those aged <50 years have significant benefits from neoadjuvant chemoradiotherapy combined with immunotherapy. Clinical T staging and magnetic resonance imaging-detected tumor regression grade after neoadjuvant therapy have predictive value for patients with pCR .
6.Efficacy analysis of robotic versus laparoscopic-assisted right hemicolectomy
Bang LIU ; Peiyao WANG ; Zhaoxiong ZHANG ; Daohan WANG ; Wenxin ZHANG ; Pengyu CHEN ; Hengbin ZHAO ; Yongjia YAN ; Weihua FU
Chinese Journal of Digestive Surgery 2025;24(4):521-527
Objective:To investigate the short-term efficacy of robotic versus laparoscopic-assisted right hemicolectomy.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopathological data of 99 patients of right colon cancer who were admitted to Tianjin Medical University General Hospital from January 2020 to December 2023 were collected. There were 50 males and 49 females, aged 69(range, 26?89)years. Of the 99 patients, 41 patients undergoing robotic-assisted right hemicolectomy were divided into the robotic group, and 58 patients undergoing laparoscopic-assisted right hemicolectomy were divided into the lapa-roscopic group. Patients received robotic-assisted or laparoscopic-assisted right hemicolectomy operated by the same major surgeon. Observation indicators:(1) propensity score matching status and com-parison of clinical data of patients between the two groups after matching; (2) intraoperative and postoperative conditions.Comparison of measurement data with normal distribution between groups was conducted using the independent sample t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the nonparametic rank sum test. Propensity score matching was performed using the 1∶1 nearest neighbor matching method. The caliper value was set as 0.1. Results:(1) Propensity score matching status and comparison of clinical data of patients between the two groups after matching. Of the 99 patients, 82 patients were successfully matched, with 41 cases in each of the robotic group and the laparoscopic group. After propensity score matching, the elimination of history of abdominal operation confounding bias ensured comparability. (2) Intraoperative and postoperative conditions. After propensity score matching, the operation time of the robotic group was 215(range, 130?340)minutes, the volume of intraoperative blood loss was 50(range, 10?400)mL, the number of lymph node dissected was 21(range, 5?55), the number of intensive care unit stay was 15, time to postoperative first flatus was 3(range, 1?12)days, time to postoperative first food intake was 4(range, 2?14)days, duration of postoperative hospital stay was 8(range, 5?25)days. The above indicators of the laparoscopic group were 210(range, 140?370)minutes, 50(range, 5?150)mL, 19(range, 5?34),20, 3(range, 0?9)days, 5(range, 2?10)days, 8(range, 6?17)days, respectively. There was no significant difference in the above indicators between patients of the two groups ( Z=?0.94, ?1.87, ?1.32, χ2=1.25, Z=0.13, ?0.83, ?0.65, P>0.05). There was no patient converted to open operation in the robotic group, versus 1 patient converted to open operation in the laparoscopic group, showing no significant difference between patients of the two groups ( P>0.05). There were 6 cases in the robotic group and 4 cases in the laparoscopic group with complications, showing no significant difference between the two groups ( χ2=0.46, P>0.05). Both groups of patients achieved R 0 resection and had no readmission 30 days after surgery. The hospital expense was (11.0±1.8)×10 4 yuan of the robotic group, versus (9.0±1.7)×10 4 yuan of the laparoscopic group, showing a significant difference between the two groups ( t=?5.27, P<0.05). Conclusion:Robot-assisted right hemicolectomy is non inferior to laparoscopic-assisted right hemicolectomy in safety and efficacy, but with higher hospitalization costs.
7.Evaluation of short-term outcomes of surgical intervention for severe pulmonary stenosis in infants and young children
Yong ZHANG ; Pengyu WANG ; Liang WANG ; Yiming TAN ; Fangran XIN ; Xu ZHANG ; Chunzhen ZHANG ; Zijun ZHOU ; Lihua LYV ; Minhua FANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2025;41(10):584-588
Objective:To evaluate the short-term efficacy of surgical treatment for severe pulmonary stenosis(PS) in infants and young children, and to clarify the impact of different surgical timings on the efficacy of PS treatment.Methods:A retrospective analysis was conducted on 24 infants and young children who underwent surgical treatment for severe PS at the General Hospital of Northern Theater Command, PLA, from January 1, 2020, to October 1, 2024. Among them, 13 were males and 11 were females. The average weight of the patients was(9.2±3.6) kg, the average gestational age was(39.3±1.7) weeks, and the average age was(15.0±13.5) months. Preoperative clinical symptoms and signs(e.g., cyanosis, shortness of breath), transpulmonary valve pressure gradient(TPVPG), right ventricular systolic pressure(RVSP), and Em/Am were recorded. The patients were divided into two groups based on surgical timing: the infant group(under 1 year old, n=12) and the toddler group(1-3 years old, n=12).Results:There were no deaths among all patients. The postoperative ICU stay was significantly longer in the infant group compared to the toddler group( P<0.05). Compared to preoperative values, surgical treatment significantly improved TPVPG, reduced RVSP, and enhanced right ventricular diastolic function, which stabilized by 3 months postoperatively( P<0.05). Intergroup comparisons revealed that the infant group had significantly lower TPVPG at 6 months postoperatively compared to the toddler group. Additionally, right ventricular diastolic function improved significantly in the infant group postoperatively, while no significant improvement was observed in the toddler group. Younger age and reduced right ventricular diastolic function were identified as major risk factors for prolonged mechanical ventilation(>24 hours). Conclusion:Surgical treatment for PS during infancy and early childhood is safe and effective. Comprehensive preoperative evaluation is crucial, and early surgical intervention is recommended for patients with impaired right ventricular function to improve prognosis.
8.Feasibility and safety of laparoscopic purse-string suture clamps and multi-functional seal caps for total laparoscopic radical total gastrectomy
Yawei QIAN ; Zhongyuan HE ; Fengyuan LI ; Pengyu LI ; Weizhi WANG ; Linjun WANG ; Diancai ZHANG ; Hao XU ; Zekuan XU ; Li YANG
Chinese Journal of Gastrointestinal Surgery 2025;28(8):908-915
Objective:To investigate the feasibility and safety of laparoscopic purse- string suture clamps combined with multi-functional seal caps for esophagojejunal Roux-en-Y anastomosis during total laparoscopic radical total gastrectomy (TLTG).Methods:This was a retrospective descriptive study of 42 patients with primary gastric malignancies who underwent TLTG at the First Affiliated Hospital of Nanjing Medical University that utilized laparoscopic purse-string suture clamps and multi-functional seal caps for esophagojejunal anastomosis between May, 2024 and January, 2025. The cohort included 33 males and 9 females, with a mean age of (67.7 ±9.5) years and a mean body mass index (BMI) of (23.9±2.9) kg/m 2. The American Society of Anesthesiologists (ASA) physical status classifications were I - II in 40 patients and III in 2 patients, and all patients were definitively diagnosed preoperatively via gastroscopy, dual-energy CT, and/or MRI. Tumor locations included the gastroesophageal junction (GEJ) in 28 cases (Siewert type II - III), the upper third of the stomach in 12 cases, and the middle third in 2 cases. The median distance of esophageal invasion was 1.3 cm, though in 10 cases this was ≥2 cm. Preoperative TNM staging was I-II in 17 patients and III in 25 patients. Surgical outcomes including operative time, anastomosis time, intraoperative blood loss, pathological results, and postoperative recovery were retrospectively analyzed. Results:All 42 operations were successful. The mean operative time was(212.5±26.4) minutes, and the average time from multi-functional seal cap placement to completion of the esophagojejunal anastomosis was (54.2±7.5) minutes. Mean intraoperative blood loss was (79.9±21.3) ml. Postoperative pathology confirmed R0 resection in all specimens, with a mean proximal esophageal margin distance of (2.1±1.6) cm. Furthermore, (51.9±15.1) lymph nodes on average were harvested from each patient; the mean time to oral intake was (149.5±41.4) hours; and the mean hospital stay was (11.3±5.4) days. Postoperative complications occurred in 6 patients: anastomotic leakage ( n=2), residual intra-abdominal infection ( n=1), pulmonary infection ( n=3), and Clavien-Dindo grade III or higher complications occurred in 2 patients. No recurrence, mortality, or anastomosis-related complications were observed within a median follow-up of 5.8 months (range 3.5-11.2). Conclusion:We find the application of the laparoscopic purse-string suture clamps and multi-functional seal caps for esophagojejunal anastomosis in TLTG to be safe and feasible, with satisfactory short-term outcomes.
9.Evaluation of short-term outcomes of surgical intervention for severe pulmonary stenosis in infants and young children
Yong ZHANG ; Pengyu WANG ; Liang WANG ; Yiming TAN ; Fangran XIN ; Xu ZHANG ; Chunzhen ZHANG ; Zijun ZHOU ; Lihua LYV ; Minhua FANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2025;41(10):584-588
Objective:To evaluate the short-term efficacy of surgical treatment for severe pulmonary stenosis(PS) in infants and young children, and to clarify the impact of different surgical timings on the efficacy of PS treatment.Methods:A retrospective analysis was conducted on 24 infants and young children who underwent surgical treatment for severe PS at the General Hospital of Northern Theater Command, PLA, from January 1, 2020, to October 1, 2024. Among them, 13 were males and 11 were females. The average weight of the patients was(9.2±3.6) kg, the average gestational age was(39.3±1.7) weeks, and the average age was(15.0±13.5) months. Preoperative clinical symptoms and signs(e.g., cyanosis, shortness of breath), transpulmonary valve pressure gradient(TPVPG), right ventricular systolic pressure(RVSP), and Em/Am were recorded. The patients were divided into two groups based on surgical timing: the infant group(under 1 year old, n=12) and the toddler group(1-3 years old, n=12).Results:There were no deaths among all patients. The postoperative ICU stay was significantly longer in the infant group compared to the toddler group( P<0.05). Compared to preoperative values, surgical treatment significantly improved TPVPG, reduced RVSP, and enhanced right ventricular diastolic function, which stabilized by 3 months postoperatively( P<0.05). Intergroup comparisons revealed that the infant group had significantly lower TPVPG at 6 months postoperatively compared to the toddler group. Additionally, right ventricular diastolic function improved significantly in the infant group postoperatively, while no significant improvement was observed in the toddler group. Younger age and reduced right ventricular diastolic function were identified as major risk factors for prolonged mechanical ventilation(>24 hours). Conclusion:Surgical treatment for PS during infancy and early childhood is safe and effective. Comprehensive preoperative evaluation is crucial, and early surgical intervention is recommended for patients with impaired right ventricular function to improve prognosis.
10.Expression of MCM2 and RF4 genes in liver cancer based on bioinformatics and experimental verification
Jianing GUO ; Pengyu ZHANG ; Xingfen WANG ; Yanchun QU ; Xianglian ZHANG ; Hui LI
International Journal of Biomedical Engineering 2025;48(1):77-87
Objective:To screen differentially expressed genes (DEGs) associated with liver cancer by bioinformatics analysis method, and to investigate the mechanism of the minichromosome maintenance protein 2 ( MCM2) and replication factor C subunit 4 ( RFC4) genes in liver cancer in vitro. Methods:Gene expression profiling data of 80 and 36 hepatocellular carcinoma tissues and 307 and 13 cirrhotic tissues were obtained from the GSE25097 and GSE98620 datasets of the gene expression analysis (GEO) database, respectively. Gene expression profiling data of 374 liver cancer tissues and 50 normal liver tissues were downloaded from the cancer genome atlas (TCGA) database. Limma and DESeqs R software were used to process the gene expression profiling data, construct protein-protein interaction networks, and analysis the relevance of these genes to survival. Weighted gene co-expression network analysis was performed to screen out the core genes. Liver cancer SMMC7721 cells were transfected with MCM2 blank plasmid (MCM2 control group), MCM2 overexpression plasmid [MCM2 WT1 group, MCM2 WT2 (2-fold WT1) group], RFC4 blank plasmid (RFC4 control group), and RFC4 overexpression plasmid [RFC4 WT1 group, RFC4 WT2 (2-fold WT1) group], respectively. The expression of MCM2 and RFC4 in liver cancer cell lines and their transfection levels were detected by real-time fluorescence quantitative reverse transcription PCR and Western blotting. The effects of MCM2 and RFC4 on the proliferation of hepatocellular carcinoma cells were detected by MTT assay and cell cloning assay, respectively. The effects of MCM2 and RFC4 on the migration of liver cancer cells were determined by the scratch assay. The effects of MCM2 and RFC4 on liver cancer cell invasion were detected by Transwell assay.Results:By bioinformatic analysis, 9 HCC DEGs were selected, including ubiquitin conjugating enzyme E2 T ( UBE2T), aurora kinase A ( AURKA), targeting protein for Xklp2 ( TPX2), MCM2, RFC4, ribonucleoside-diphosphate reductase subunit M2 ( RRM2), serine peptidase inhibitory factor Kazal type 1 ( SPINK1), collagen type XV alpha 1 chain ( COL15A1) and C-C motif chemokine 25 ( CCL25). Among the six genes associated with clinical stages, the MCM2 and RFC4 genes were found to be strongly associated with prognosis in liver cancer. The relative protein expression of MCM2 and RFC4 in HepG2 cells (1.83±0.07, 1.44±0.09) and SMMC7721 cells (1.74±0.05, 1.43±0.08) was higher than that in MIHA cells (1.00±0.02, 1.00±0.03), and all the differences were statistically significant (all P<0.05). The relative gene expression of MCM2 and RFC4 in HepG2 cells (14.30±0.12, 5.10±0.18) and SMMC7721 cells (10.60±0.11, 7.60±0.07) was higher than that in MIHA cells (1.00±0.05, 1.00±0.03), and all the differences were statistically significant (all P<0.05). Compared with the MCM2 control group, the absorbance values [(0.28±0.01 and 0.21±0.01) vs 0.18±0.03], the number of clonal cells [(717±12 and 782±29) cells vs (389±17) cells], the percentage migration [(0.43±0.02 and 0.68±0.01) vs 0.15±0.06], and the number of cellular invasions [(933±21 and 821±11) cells vs (409±16) cells] were higher in the MCM2 WT1 and MCM2 WT2 groups, and the differences were all statistically significant (all P<0.05). Compared with the RFC4 control group, the absorbance values [(0.30±0.02 and 0.21±0.01) vs 0.17±0.02], the number of cloned cells [(571±11 and 728±9) cells vs (373±23) cells], the percentage migration [(0.75±0.11 and 0.67±0.04) vs 0.34±0.07], and the number of cell invasion [(835±26 and 818±18) cells vs (629±12) cells] were higher in the RFC4 WT1 and the RFC4 WT2 groups, and the differences were statistically significant (all P<0.05). Conclusions:MCM2 and R FC4 genes play a role in promoting tumorigenesis and growth in hepatocellular carcinoma.

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