摘要
目的探讨微小RNA(miR)-497-5p对子宫内膜癌Ishikawa细胞增殖、侵袭和凋亡的影响及其机制。方法将体外培养的Ishikawa细胞分为Ctrl组(未处理)、miR-NC组(转染模拟物阴性对照)和mi R-497-5p组(转染mi R-497-5p模拟物),采用实时荧光定量PCR检测Ishikawa细胞中mi R-497-5p表达,噻唑蓝(MTT)实验、Transwell小室实验和流式细胞仪分别检测Ishikawa细胞增殖、侵袭和凋亡情况,免疫印迹法(Westernblot)检测Ishikawa细胞中Wnt3a、-连环蛋白(-catenin)、磷酸化(p)--catenin、c-myc和细胞周期素D1(cyclinD1)蛋白表达情况,双荧光素酶报告基因实验检测mi R-497-5p和Wnt3a的靶向关系。结果与Ctrl组和miR-NC组比较,mi R-497-5p组Ishikawa细胞中miR-497-5p表达水平、细胞凋亡率和p--catenin表达水平明显升高,而细胞增殖能力、侵袭能力和Wnt3a、-catenin、c-myc、cyclinD1蛋白表达水平明显降低(P<0.05);与Ctrl组比较,miR-NC组中上述各指标差异均无统计学意义。双荧光素酶报告基因实验证实Wnt3a是mi R-497-5p的靶基因。结论 miR-497-5p可抑制子宫内膜癌Ishikawa细胞增殖、侵袭并诱导细胞凋亡,其作用机制可能与其靶向调控Wnt3a/β-catenin信号通路有关。
Objective To investigate the effects of microRNA(miR)-497-5 p on proliferation, invasion and apoptosis of endometrial carcinoma Ishikawa cells and its mechanism. Methods Ishikawa cells cultured in vitro were divided into three groups: Ctrl group(untreated), mi R-NC group(transfected with mimic negative control) and mi R-497-5 p group(transfected with miR-497-5 p mimic), the expression of miR-497-5 p in Ishikawa cells was detected by real-time fluorescent quantitative PCR, the proliferation, invasion and apoptosis of Ishikawa cells were detected by methyl thiazolyl tetrazolium(MTT) assay, Transwell chamber test and flow cytometry, Western blot was used to detect the protein expressions of Wnt3 a, β-catenin, phosphorylated(p)-β-catenin, c-myc and cyclinD1 in Ishikawa cells, in addition, double luciferase reporter gene assay was used to detect the targeting relationship between miR-497-5 p and Wnt3 a. Results Compared with those in Ctrl group and miR-NC group, the miR-497-5 p expression level, apoptosis rate and p-β-catenin expression level were significantly higher in miR-497-5 p group, however, cell proliferation, invasion and protein expression levels of Wnt3 a, β-catenin, c-myc and cyclinD1 were significantly lower(P<0.05);compared with those in the Ctrl group, there was no significant difference in the above indexes in the miR-NC group. In addition, double luciferase reporter gene assay confirmed that Wnt3 a was the target gene of miR-497-5 p. Conclusion MiR-497-5 p can inhibit the proliferation, invasion, and induce the apoptosis of endometrial carcinoma Ishikawa cells, and its mechanism may be related to its targeted regulation of Wnt3 a/β-catenin signaling pathway.
作者
林娟
闫咨儒
高艳宇
LIN Juan;YAN Ziru;GAO Yanyu(Department of Gynecology and Obstetrics,Chengdu Women and Children's Central Hospital,Chengdu,Sichuan 610000,China;Department of Obstetrics,Chengdu Women and Children's Central Hospital,Chengdu,Sichuan 610000,China)
出处
《中国优生与遗传杂志》
2021年第3期345-349,共5页
Chinese Journal of Birth Health & Heredity
