miR-96在子宫内膜癌中表达的变化及其对癌细胞恶性生物学行为的影响

Expression change of miR-96 in endometrial cancer and its effect on malignant biological behaviors of cancer cells

目的:探讨miR-96在子宫内膜癌组织和细胞中表达的变化对瘤细胞恶性表型的影响及其可能的作用机制。方法:选取2016年4月至2018年12月在我院妇产科接受手术治疗的76例子宫内膜癌患者的癌组织标本,采用实时荧光定量PCR(q PCR)检测人子宫内膜癌组织和细胞中miR-96的表达情况,并分析miR-96的表达与患者临床病理特征的相关性。体外转染miR-96 inhibitor至人子宫内膜癌Ishikawa细胞中,q PCR检测Ishikawa细胞中miR-96的表达变化,分别应用CCK-8实验、克隆形成实验、流式细胞术、划痕实验和Transwell实验检测Ishikawa细胞恶性生物学行为的变化,WB检测Ishikawa细胞中FOXO1蛋白的表达变化,双荧光素酶报告基因观察miR-96和FOXO1的靶向关系。结果:qPCR结果显示,miR-96在人子宫内膜癌细胞JEC、Ishikawa、HEC-1B和子宫内膜癌组织中异常高表达﹙均P<0.01﹚,并且miR-96表达与患者FIGO分期和淋巴结转移密切关联(均P<0.05)。转染miR-96 inhibitor后,Ishikawa细胞miR-96表达水平明显降低(P<0.01),其增殖活性和克隆形成能力明显下降(均P<0.01)、凋亡率明显上升(P<0.01)、划痕愈合率和侵袭穿膜细胞数明显下降(P<0.01)。双荧光素酶报告基因显示miR-96可以直接靶向FOXO1,WB结果显示miR-96能够负向调控Ishikawa细胞FOXO1蛋白的表达(P<0.01)。结论:miR-96在子宫内膜癌组织和细胞中异常高表达,抑制miR-96表达能够抑制子宫内膜癌细胞增殖、迁移及侵袭并促进其凋亡,其机制可能与靶向调控FOXO1有关。

Objective: To investigate the expression changes of miR-96 in endometrial cancer tissues and cells, and to explore its effect on tumor malignant phenotypes as well as the possible mechanisms. Methods: From April 2016 to December 2018, 76 cases of endometrial cancer tissues from 76 patients who were surgically treated in the Department of Obstetrics and Gynecology of our hospital were selected for this study. q PCR was used to detect the expression of miR-96 in human endometrial cancer tissues and cells, and the correlation between the miR-96 expression and the clinicopathological characteristics of patients was analyzed. miR-96 inhibitor was transfected into human endometrial cancer Ishikawa cells in vitro. After transfection, the expression of miR-96 in Ishikawa cells was detected by q PCR;the tumor biological behaviors of Ishikawa cells were detected by CCK-8 test, Clone formation test, Flow cytometry,Scratch test and Transwell test;and the FOXO1 protein expression in Ishikawa cells was detected by WB. At the same time, Dual luciferase reporter gene assay was used to observe the targeting relationship between miR-96 and FOXO1. Results: The results of qPCR showed that the expression of miR-96 was abnormally high in human endometrial cancer cells(JEC, Ishikawa, HEC-1 B) and endometrial cancer tissues(all P<0.01), and the expression of miR-96 was closely related to FIGO stage and lymph node metastasis(all P<0.05). After transfection with miR-96 inhibitor, the expression level of miR-96 in Ishikawa cells decreased significantly(P<0.01),the proliferation activity and clone formation ability decreased significantly(all P<0.01), the apoptotic rate increased significantly(P<0.01), and the scratch healing rate and the number of invasive transmembrane cells decreased significantly(P<0.01). Dual luciferase reporter gene assay showed that miR-96 could directly target FOXO1, and WB showed that miR-96 could negatively regulate FOXO1 protein expression in Ishikawa cells(P<0.01). Conclusion: The expression of miR-96 is abnormally high in endometrial cancer tissues and cells. Inhibiting the expression of miR-96 can inhibit the proliferation, invasion and migration of endometrial cancer cells and promote their apoptosis. The mechanism may be related to the targeted regulation of FOXO1.