miR-25-3p和ZEB2在宫颈癌中表达的临床意义及对宫颈癌细胞增殖和侵袭的作用探讨

Clinical significance of miR-25-3p and ZEB2 expression in cervical cancer and their effects on proliferation and invasion of cervical cancer cells

背景微小RNA-25-3p(microRNA-25-3p,miR-25-3p)是与肿瘤密切相关的miRNA,锌指E盒结合的同源盒蛋白2(zinc finger E-box binding homeobox 2,ZEB2)是miR-25-3p的靶基因,而miR-25-3p能否通过靶向ZEB2在宫颈癌进展中发挥作用尚不清楚。目的探讨miR-25-3p、ZEB2在宫颈癌组织中表达的临床意义及对宫颈癌细胞增殖和侵袭的作用。方法选取四川大学华西三亚医院2013年1月-2014年12月治疗的宫颈癌患者84例,获取病理组织,另收集因良性疾病切除的正常宫颈组织40例作为对照,检测宫颈癌组织和正常宫颈组织中miR-25-3p、ZEB2 mRNA及蛋白的表达情况;分析miR-25-3p、ZEB2 mRNA及蛋白的表达与宫颈癌患者临床病理参数及预后的关系。Pearson线性相关分析miR-25-3p与ZEB2 mRNA及蛋白在宫颈癌组织中表达的相关性。常规培养宫颈癌细胞HeLa,分为NC组、miR-25-3p mimic组和miR-25-3p+oe-ZEB2组,MTS实验检测各组细胞的增殖能力,Boyden实验检测各组细胞的侵袭能力。结果miR-25-3p在宫颈癌组织中的表达显著低于其在正常宫颈组织中的表达(P<0.05),ZEB2 mRNA及蛋白在宫颈癌组织中的表达显著高于其在正常宫颈组织中的表达(P<0.05)。miR-25-3p在宫颈癌组织中的表达水平与淋巴结转移、肌层浸润深度和FIGO分期相关(P<0.05)。ZEB2 mRNA及蛋白在宫颈癌组织中的表达水平与组织学分级、淋巴结转移和FIGO分期相关(P<0.05)。Kaplan-Meier生存曲线显示,miR-25-3p低表达组宫颈癌患者5年总生存率低于miR-25-3p高表达组患者(P<0.05),ZEB2 mRNA及蛋白高表达的宫颈癌患者5年总生存率低于ZEB2低表达组患者(P<0.05)。宫颈癌组织中miR-25-3p与ZEB2 mRNA及蛋白的表达呈负相关(P<0.05)。Target Scan7.1软件预测及双荧光素酶报道基因实验结果显示ZEB2为miR-25-3p的靶基因。与NC组相比,miR-25-3p mimic组和miR-25-3p+oe-ZEB2组细胞增殖和侵袭能力降低(P<0.05);与miR-25-3p mimic组相比,miR-25-3p+oe-ZEB2组细胞增殖和侵袭能力增加(P<0.05)。结论miR-25-3p、ZEB2与宫颈癌患者的不良病理参数和预后相关,miR-25-3p靶向负调控ZEB2抑制宫颈癌的增殖和侵袭。miR-25-3p和ZEB2有可能成为宫颈癌预后新的标志物和治疗靶标。

Background MicroRNA-25-3p(miR-25-3p)is a miRNA that is closely related to tumors.Zinc finger E-box binding homeobox 2(ZEB2)is the target gene of miR-25-3p.However,whether miR-25-3p can play a role in the progression of cervical cancer by targeting ZEB2 is unclear.Objective To investigate the clinical significance of miR-25-3p and ZEB2 expression in cervical cancer tissues and its effect on the proliferation and invasion of cervical cancer cells.Methods A total of 84 patients with cervical cancer treated in West China(Sanya)Hospital of Sichuan University from January 2013 to December 2014 were selected.Another 40 cases of normal cervical tissues excised due to benign diseases were collected as the control.The mRNA and protein expression levels of ZEB2 and miR-25-3p in cervical cancer tissues and normal cervical tissues were detected.The relationship between the mRNA and protein expression levels of ZEB2 and miR-25-3p and the clinicopathological parameters and prognosis of cervical cancer patients was analyzed.Pearson linear correlation analysis was used to detect the correlation between miR-25-3p,mRNA and protein expression in cervical cancer tissues;HeLa cells were routinely cultured,divided into NC group,miR-25-3p mimic group and miR-25-3p+oe-ZEB2 group.MTS experiment was used to detect the proliferation ability of each group,and Boyden experiment was used to detect the invasion ability of each group.Results The expression level of miR-25-3p in cervical cancer tissues was significantly lower than that in the normal cervical tissues(P<0.05),and the expression levels of ZEB2 mRNA and protein in cervical cancer tissues were significantly higher than those in normal cervical tissues(P<0.05).The expression level of miR-25-3p in cervical cancer tissues was correlated with lymph node metastasis,depth of myometrial invasion and FIGO stage(P<0.05).The expression levels of ZEB2 mRNA and protein in cervical cancer tissues were correlated with histological grade,lymph node metastasis and FIGO stage(P<0.05).Kaplan-Meier survival curve showed that the 5-year OS rate of cervical cancer patients in the miR-25-3p low expression group was lower than that in the miR-25-3p high expression group(P<0.05),and the 5-year OS rate of cervical cancer patients with high ZEB2 mRNA and protein expression level was lower than that of ZEB2 low expression group(P<0.05).The expression levels of miR-25-3p and ZEB2 in cervical cancer tissues were negatively correlated(P<0.05).Target Scan7.1 software prediction and dual luciferase reporter gene experiment results showed that ZEB2 was the target gene of miR-25-3p.Compared with the NC group,the cell proliferation and invasion ability of the miR-25-3p mimic group and miR-25-3p+oe-ZEB2 group were reduced(P<0.05).Compared with the miR-25-3p mimic group,the cell proliferation and invasion ability of miR-25-3p+oe-ZEB2 group were increased(P<0.05).Conclusion miR-25-3p and ZEB2 are related to the adverse patho-logical parameters and prognosis of cervical cancer patients,and miR-25-3p negatively regulates ZEB2 to inhibit the proliferation and invasion of cervical cancer.miR-25-3p and ZEB2 may be new prognostic markers and therapeutic targets for cervical cancer.