1.Expression of I kappa B kinase-interacting protein in cervical cancer tissue and its effect on proliferation,migration and invasion of cervical cancer cells
Yan WANG ; Zouyu ZHAO ; Panpan YU ; Ping YANG
Journal of Jilin University(Medicine Edition) 2025;51(2):341-351
Objective:To explore the correlation between IκB kinase-interacting protein(IKBIP)expression in tumor cells within cervical cancer tissues(TCCCT)and the clinical pathological characteristics and prognosis of patients,as well as its impact on the biological behaviors of cervical cancer(CC)cells HeLa and SiHa,and to elucidate its potential mechanism.Methods:GENT2 and Kaplan-Meier plotter databases were utilized to analyze the differential expression of IKBIP mRNA in CC and normal cervical tissues,as well as its correlation with the clinical prognosis of CC patients.The Hallmark reference gene set was chosen for pathway enrichment analysis using the Gene Set Enrichment Analysis(GSEA)software.Immunohistochemistry method was used to detect the IKBIP protein expression in TCCCT and epithelial cells in normal cervical tissue(ECNCT),and analyze the correlations between its expression level and clinicopathological characteristics and prognosis of CC patients.Furthermore,univariate and multivariate Cox regression analyses were performed to identify the risk factors impacting the prognosis of CC patients.The stably transfected cells of CC(HeLa cells and SiHa cells)with IKBIP knockdown were established for the experiment,which were divided into sh-NC group and sh-IKBIP group.The expression of IKBIP protein in various groups was assessed using Western blotting method.The cell proliferation activity and the percentage of 5-ethynyl-2'-deoxyuridine(EdU)positive cells were measured using cell counting kit-8(CCK-8)and EdU methods,while Transwell chamber assay was employed to determine the numbers of migration and invasion cells in various groups.Additionally,the expression levels of E-cadherin,N-cadherin,and Snail proteins in the cells in various groups were analyzed by Western blotting method.Results:The GENT2 database analysis revealed that the expression level of IKBIP mRNA in CC tissue was higher than that in normal cervical tissue(P<0.001).The GSEA enrichment analysis identified the epithelial-mesenchymal transition(EMT)pathway as the top-ranked pathway in IKBIP high-expression group.The immunohistochemistry results indicated the positive expression rate of IKBIP protein in TCCCT was higher than that in ECNCT(50.5%vs 8.0%),and its over-expression was associated with FIGO stage(2018)and differentiation grade of tumor(P<0.05).The univariate and multivariate Cox regression analyses suggested that lymph node metastasis(LNM)and high expression of IKBIP were the risk factors affecting the overall survival(OS)and progression-free survival(PFS)of CC patients(P<0.05).The Western blotting method results showed that compared with sh-NC group,the expression level of IKBIP protein in the cells in sh-IKBIP group was decreased(P<0.05).The CCK-8 and EdU assay results showed that compared with sh-NC group,the proliferation activity and the percentage of EdU positive cells in sh-IKBIP group were decreased(P<0.05).The Transwell chamber assay results showed that compared with sh-NC group,the numbers of migration and invasion cells in sh-IKBIP group were significantly decreased(P<0.05).Additionally,the Western blotting method results indicated that compared with sh-NC group,the expression level of E-cadherin protein in sh-IKBIP group in CC cells was increased(P<0.05),while the expression levels of N-cadherin and Snail protein were decreased(P<0.05).Conclusion:The expression of IKBIP protein is significantly up-regulated in HeLa and SiHa cells derived from CC,and it is closely correlated with poor prognosis in CC patients.Suppression of IKBIP protein expression can effectively inhibit the proliferation,invasion and migration capabilities as well as EMT process of CC cells.
2.The value of six tumor markers in preoperative diagnosis and prognostic evaluation of advanced serous ovarian cancer
Jishuai HOU ; Zouyu ZHAO ; Yan WEI ; Rui BAI ; Ping YANG
Journal of Chinese Physician 2024;26(1):36-42
Objective:To explore the diagnostic and prognostic value of six tumor markers, namely carbohydrate antigen 125 (CA125), carcinoembryonic antigen (CEA), alpha fetoprotein (AFP), carbohydrate antigen 153 (CA153), carbohydrate antigen 199 (CA199), and carbohydrate antigen 724 (CA724), in the pathological types of epithelial ovarian cancer (EOC).Methods:A retrospective analysis was conducted on the preoperative tumor markers and clinical pathological data of 131 EOC patients admitted to the First Affiliated Hospital of Shihezi University from January 2010 to May 2022, and follow-up was conducted. Patients were divided into high-grade serous ovarian cancer (HGSOC) group and other groups (mucinous cancer, endometrioid carcinoma, clear cell carcinoma, mixed carcinoma) according to pathological type and tumor grade. We investigated the correlation between the levels of six tumor markers and the pathological types of EOC. By drawing receiver operating characteristic (ROC) curves and comparing the area under the curve (AUC), we also investigated the diagnostic value of single and combined detection of six tumor markers for the pathological types of EOC. K-M survival analysis was used to explore the impact of tumor markers on patient prognosis.Results:The levels of CA125 and CA153 in the HGSOC group were significantly higher than those in other pathological groups ( Z=-2.571, -5.416, all P<0.05); CA153 had good diagnostic performance for HGSOC (AUC=0.777), and the combined detection of CA125+ CA153, CA153+ AFP, CA153+ CA199, CA153+ CA724, CA153+ CE had better diagnostic performance than the single detection of CA125 and CA153 (AUC=0.781, 0.784, 0.809, 0.803, 0.773). Among them, the combined detection of CA125, CA153, and CA199 had the best diagnostic performance (AUC=0.816, Youden′s index 0.532); the elevated levels of CA153 and CA153+ CA199 indicated poor recurrence free survival (PFS) in patients (all P<0.05), while the elevated levels of CA153+ CA199 was an independent risk factor for recurrence in patients ( P=0.022); the elevated levels of CA153+ CA199 and CA125+ CA153+ CA199 indicated poorer OS in patients (all P<0.05). Conclusions:The serum levels of CA125 and CA153 are elevated in patients with HGSOC. Elevated levels of CA153, CA153+ CA199, and CA125+ CA153+ CA199 are associated with poor prognosis in patients. The combined detection of CA125, CA153, and CA199 has the best diagnostic efficacy and can serve as a potential biomarker for assisting in the diagnosis of HGSOC and evaluating patient prognosis.
3.Correlation analysis of GSTO1 expression in cervical cancer tissue with prognosis and the impact of N-glycosylation on the malignant biological behavior of cervical cancer
Panpan YU ; Ping YANG ; Qianyu SUN ; Weirui GAO ; Zouyu ZHAO ; Chongfeng SUN
Acta Universitatis Medicinalis Anhui 2023;58(12):2002-2010
Objective To investigate the expression of(glutathione S-Transferase Omega-1,GSTO1)in cervical cancer tissue and its correlation with patient survival time,and to explore the impact of GSTO1 N-glycosylation on proliferation,migration,invasion,and epithelial-mesenchymal transition of cervical cancer.Methods By using immunohistochemistry,the expression levels of GSTO1 in tumor cells of 82 cervical cancer patients were detected,and the correlation between GSTO1 expression and clinical pathological features was analyzed.Kaplan-Meier meth-od was used to plot survival curves and evaluate the impact of GSTO1 expression in cervical cancer tissues on pa-tient survival time.Univariate and multivariate Cox regression analyses were performed to assess the independent prognostic factors influencing cervical cancer prognosis.The NetNGlyc 1.0 Server database predicted potential N-glycosylation modification sites of GSTO1(Asn55,Asnl35,Asn190).The cervical cancer cells(HeLa)were transfected with GSTO1 N-glycosylation site mutation vectors at positions 55,135,and 190,as well as GSTO1 wild-type vector and empty vector.Stable transfected cells were selected using puromycin.Western blot experi-ments were performed to assess the effectiveness of lentiviral interference.The effects of GSTO1 N-glycosylation site mutations on proliferation,migration,and invasion of HeLa cells were evaluated using EdU proliferation assay,wound healing assay,and Transwell assay.The effect of GSTO1 N-glycosylation site mutations on the epithelial-mesenchymal transition of HeLa cells was detected using the Western blot experiment.Results Immunohistochem-istry results revealed high expression of GSTO1 in cervical cancer tissues.The expression rate of GSTO1 was signifi-cantly higher in cervical cancer tissues with deep stromal invision≥1/2,lymphovascular space invasion,and lymph node metastasis(P<0.05).Moreover,high expression of GSTO1 was associated with poorer overall surviv-al.After N-glycosylation site-specific mutation of GSTO1,the cell count of proliferation,migration,and invasion in HeLa cells significantly decreased(P<0.05).The Western blot results showed that N-glycosylation site mutation of GSTO1 significantly inhibited the epithelial-mesenchymal transition of HeLa cells.Conclusion The expression of GSTO1 in cervical cancer tissues is associated with stromal infiltration depth,lymphovascular space invasion and lymph node metastasis,and it is also correlated with shorter patient survival time.Site-specific mutations in GSTO1 N-glycosylation significantly inhibit the proliferation,migration and epitheli al-mesenchymal transition of HeLa cells.

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