卵巢癌中REG4的表达及其作用机制

The clinicopathological significance and molecular mechanism of REG4 expression in ovarian cancer

目的探讨卵巢癌中再生基因4(REG4)表达的临床病理意义及其作用机制。方法利用UCSC Xena数据库分析卵巢癌中REG4 mRNA及相关基因的表达情况;采用Kaplan-Meier Plotter分析卵巢癌中REG4 mRNA表达与生存的关系;稳定转染pcDNA3.1-REG4真核表达质粒和pcDNA3.1空载体至SKOV3卵巢癌细胞,采用实时PCR和Western blotting筛选REG4过表达细胞株;通过MTT法、流式细胞术、Transwell法分别检测REG4对卵巢癌细胞增殖、凋亡和侵袭的影响;应用TCGA数据库对REG4表达的卵巢癌进行基因富集分析。结果与正常卵巢组织相比,复发性卵巢癌中REG4 mRNA表达最高(P<0.05),卵巢癌中次之(P<0.05),REG4高表达与卵巢癌的总体生存率、无进展生存率和后进展生存率降低相关(P<0.05);REG4过表达可抑制SKOV3细胞凋亡并促进SKOV3细胞增殖和侵袭(P<0.05);REG4与凋亡相关基因呈负相关富集(P<0.05),与药物代谢细胞色素P450相关基因呈正相关富集(P<0.05)。结论卵巢癌中REG4表达上调,且与卵巢癌不良预后相关,它通过调节细胞的增殖、凋亡和侵袭促进卵巢癌的发生发展,可作为卵巢癌预后评估和基因治疗的分子靶标。

Objective To investigate the clinicopathological significance and the effects of regenerating gene family member 4(REG4)expression in ovarian cancer.Methods REG4 mRNA expression in ovarian cancer cells was analyzed using the data downloaded from the UCSC Xena database.Kaplan-Meier Plotter was used to perform the survival analysis.SKOV3 cells were transfected with the REG4-expressing plasmid,and a mock sample was prepared by transfecting SKOV3 cells with pcDNA3.1 plasmid.The effects of REG4 expression on cell proliferation,cell apoptosis,and cell invasion were assessed using MTT,flow cytometry,and Transwell assays,respectively.Gene set enrichment analysis was performed to analyze the relationship between REG4 expression and cell-signaling pathways.Results Gene expression analysis revealed that REG4 expression was highest in the recurrent ovarian cancer,followed by the primary ovarian cancer,and lowest in the normal ovary tissues(P<0.05).Survival analysis revealed that the overall,progression-free,and post-progression survival rates of ovarian cancer patients with high REG4 expression levels were significantly lower than those with low REG4 expression levels(P<0.05).REG4 overexpression suppressed apoptosis and promoted cell proliferation and invasion in the ovarian cancer cells when compared to the mock sample(P<0.05).The cell-signaling pathways that were found to be related to REG4 expression in ovarian cancer included apoptosis,drug metabolism,and cytochrome P450(P<0.05).Conclusion In this study,REG4 overexpression was shown to play a crucial role in ovarian carcinogenesis.High expression of REG4 was correlated with poor prognosis in ovarian cancer patients.REG4 was found to promote tumorigenesis and subsequent progression of ovarian cancer by regulating cell proliferation,apoptosis,and invasion.Thus,REG4 can be used as a molecular target for the estimation of prognosis and for designing gene therapy for ovarian cancer.