子宫内膜癌组织中GMNN、RFC4表达变化及其与患者预后的关系

Expression changes of GMNN and RFC4 in endometrial carcinoma tissues and their relationship with the prognosis of patients

目的分别从生物信息学、组织标本的角度观察人子宫内膜癌组织中Geminin(GMNN)及人类复制因子C第四大亚基(RFC4)的表达变化,分析其与患者预后的关系。方法利用UALCAN数据库分析GMNN、RFC4 mRNA和蛋白在子宫内膜癌组织和正常子宫内膜组织中的表达差异,利用OncoLnc数据库比较GMNN、RFC4 mRNA高表达与低表达子宫内膜癌患者的总生存率,利用GEPIA数据库分析GMNN与RFC4mRNA表达的相关性。取手术切除的40例子宫内膜癌患者的新鲜癌组织和癌旁组织标本,采用RT-qPCR法检测GMNN、RFC4 mRNA表达。取正常子宫内膜组织120例、不典型增生子宫内膜组织100例、子宫内膜癌组织133例石蜡标本,采用免疫组化EnVision法检测GMNN、RFC4蛋白表达,比较不同临床病理参数的子宫内膜癌患者肿瘤组织GMNN、RFC4蛋白表达的差异。Kaplan-Meier生存分析法比较不同GMNN、RFC4蛋白表达水平患者的总生存率,Cox回归分析法分析影响子宫内膜癌患者预后的相关因素;Pearson相关分析法分析GMNN、RFC4蛋白在子宫内膜癌组织中表达的相关性。结果生物信息学分析显示,GMNN、RFC4 mRNA和蛋白在人子宫内膜癌中的表达均高于正常子宫内膜组织(P均<0.05);GMNN及RFC4 mRNA高表达患者的生存时间均低于低表达患者(P均<0.05);GMNN与RFC4 mRNA表达呈正相关(r=0.58,P <0.01)。组织标本检测显示,GMNN、RFC4 mRNA在子宫内膜癌组织中的表达均高于癌旁组织(P均<0.05);GMNN、RFC4蛋白在子宫内膜癌中的高表达率高于正常子宫内膜组织和不典型子宫内膜组织(P均<0.05),合并淋巴结转移、FIGO病理分期Ⅲ+Ⅳ期、Ki67表达阳性、p53表达阳性患者的GMNN、RFC4蛋白高表达率分别高于无淋巴结转移、FIGO病理分期Ⅰ+Ⅱ期、Ki67表达阴性、p53表达阴性的患者(P均<0.05)。GMNN、RFC4蛋白高表达患者的总生存率低于低表达患者(P均<0.05),GMNN蛋白高表达、RFC4蛋白高表达、淋巴结转移、FIGO病理分期Ⅲ+Ⅳ期、Ki-67阳性是子宫内膜癌患者预后的独立影响因素(P均<0.05)。GMNN与RFC4蛋白在子宫内膜癌组织中的表达呈正相关(rs=0.303,P <0.01)。结论 GMNN、RFC4mRNA及蛋白在子宫内膜癌组织中表达升高,其表达水平与子宫内膜癌的恶性生物学行为、预后密切相关,两者可能作为子宫内膜癌诊断和治疗的新靶点。

Objective To observe the expression of Geminin(GMNN) and the fourth subunit of human replicator C(RFC4) in human endometrial carcinoma from the perspectives of bioinformatics and tissue specimens,and to analyze their relationship with the prognosis of patients.Methods UALCAN database was used to analyze the expression differences of GMNN and RFC4 mRNA and protein between the endometrial cancer tissues and normal endometrial tissues,and the overall survival rates of patients with high and low GMNN and RFC4 gene expression was compared using Onco Lnc database.GEPIA database was used to analyze the correlation between GMNN and RFC4 gene.The expression of GMNN and RFC4 mRNA was detected by RT-qPCR in fresh and adjacent tissues of 40 patients with endometrial carcinoma.Paraffin samples were collected from 120 normal endometrial tissues,100 atypical hyperplasia endometrial tissues,and 133 endometrial cancer tissues.Immunohistochemical En Vision method was used to detect GMNN and RFC4 protein expression,and the differences of GMNN and RFC4 protein expression in tumor tissues of endometrial cancer patients with different clinicopathological parameters were compared.Kaplan-meier survival analysis was used to compare the overall survival rates of patients with different GMNN and RFC4 protein expression levels,and Cox regression analysis was used to analyze the prognostic factors of endometrial cancer patients.The correlation between GMNN and RFC4 protein expression in endometrial carcinoma tissues was analyzed by Pearson correlation analysis.Results Bioinformatics analysis showed that the mRNA and protein expression levels of GMNN and RFC4 were higher in the human endometrial carcinoma tissues than in the normal endometrial tissues(all P < 0.05);the survival time of patients with high GMNN and RFC4 gene expression levels was lower than that of patients with low expression levels(all P < 0.05);GMNN was positively correlated with RFC4 gene expression(r = 0.58,P < 0.01).The expression levels of GMNN and RFC4 mRNA were higher in the endometrial carcinoma tissues than in the para-carcinoma tissues(all P < 0.05);the high expression rates of GMNN and RFC4 protein in the endometrial carcinoma were higher than those in normal and atypical endometrial tissues(all P < 0.05),and the high expression rates of GMNN and RFC4 protein in patients with lymph node metastasis,FIGO pathological stage Ⅲ + Ⅳ,positive Ki67 expression,and positive p53 expression were higher than those in patients without lymph node metastasis,FIGO pathological stageⅠ + Ⅱ,negative Ki67 expression,and negative p53 expression(all P < 0.05).The overall survival rate of patients with high expression of GMNN and RFC4 protein was lower than that of patients with low expression of GMNN and RFC4 protein(P < 0.05);high expression of GMNN protein,high expression of RFC4 protein,lymph node metastasis,FIGO pathological stage Ⅲ + Ⅳand positive Ki-67 were independent prognostic factors of endometrial carcinoma(all P < 0.05).GMNN was positively correlated with the expression of RFC4 protein in endometrial carcinoma(rs= 0.303,P < 0.01).Conclusion The mRNA and protein expression levels of GMNN and RFC4 increase in the endometrial cancer tissues,and their expression levels are closely related to the malignant biological behavior and prognosis of endometrial cancer patients,which may be used as new targets for diagnosis and treatment of endometrial cancer.