LncRNA XIST在子宫内膜癌组织中的表达及其靶向microRNA-101-3p对癌细胞生物行为学的影响

Expression of lncRNA XIST in endometrial carcinoma and effect of targeting microRNA-101-3p on biological behavior of cancer cells

摘要:

目的 探究lncRNA XIST在子宫内膜癌组织中的表达及其靶向microRNA-101-3p（miR-101-3p）对癌细胞生物行为学的影响。方法 选取2019年7月—2021年7月南通市妇幼保健院82例手术切除且经术后病理确诊的子宫内膜癌患者的癌组织及癌旁正常子宫内膜组织标本。采用实时荧光定量聚合酶链反应（qRT-PCR）检测子宫内膜组织lncRNA XIST、miR-101-3p mRNA的相对表达量；比较不同因素间lncRNA XIST、miR-101-3p mRNA的差异；取Ishikawa细胞随机分为对照组（C组）、空质粒组（NC组）、lncRNA XIST表达抑制组（sh-XIST组），其中NC组和sh-XIST组分别转染空载质粒与plko-lncRNA XIST-shRNA，C组不进行任何处理；CCK-8法检测细胞增殖活性；划痕实验检测细胞迁移能力；Transwell小室实验检测细胞侵袭能力；双荧光素酶报告基因观察lncRNA XIST与miR-101-3p的靶向性。结果 子宫内膜癌组织中lncRNA XIST mRNA相对表达量高于癌旁组织（P <0.05），miR-101-3p mRNA相对表达量低于癌旁组织（P <0.05）。不同TNM分期和淋巴结是否转移患者的lncRNA XIST、miR-101-3p mRNA相对表达量比较，差异有统计学意义（P <0.05）。C组、NC组、sh-XIST组在不同时间点Ishikawa细胞增殖活性的比较采用重复测量设计的方差分析，结果 ①不同时间点的Ishikawa细胞增殖活性有差异（P <0.05）；②3组的Ishikawa细胞增殖活性有差异（P <0.05）；③3组的Ishikawa细胞增殖活性变化趋势有差异（P <0.05）。sh-XIST组细胞迁移率低于C组和NC组（P <0.05）。sh-XIST组侵袭个数少于C组和NC组（P <0.05）。双荧光素酶报告基因显示，lncRNA XIST可以靶向作用于miR-101-3p。结论 lncRNA XIST和miR-101-3p mRNA在子宫内膜癌组织中异常高表达，并且在不同TNM分期及是否淋巴结转移患者子宫内膜癌组织中的表达有差异。抑制Ishikawa细胞中lncRNA XIST的表达可以抑制癌细胞的恶性细胞生物学行为，其机制可能与靶向调控miR-101-3p有关。

Abstract:

Objective To investigate the expression of lncRNA XIST in endometrial carcinoma and the effect of targeting microRNA-101-3p(miR-101-3p) on the biological behavior of cancer cells.Methods The cancer tissues of 82 patients with endometrial cancer and matched normal endometrial tissues adjacent to the cancer were taken from gynecological surgery in our hospital from July 2019 to July 2021. The expression of lncRNA Xist was detected by real-time fluorescence quantitative PCR, and the correlation between the expression of lncRNA Xist and clinicopathological parameters was analyzed. Ishikawa cells were randomly divided into control group (C group), empty plasmid group (NC group), and lncRNA Xist expression inhibition group (sh XIST group). NC group and sh XIST group were transfected with empty plasmid and plko lncRNA XIST shRNA respectively, and C group was not treated. Cell proliferation activity was detected by CCK-8. The cell migration ability was detected by scratch test. The invasion ability of cells was detected by Transwell chamber experiment. The targeting of lncRNA XIST and miR-101-3p was observed by double luciferase reporter gene.Results The expression of lncRNA XIST and miR-101-3p in endometrial carcinoma was higher than that in adjacent tissues, and the expression of lncRNA XIST and miR-101-3p was related to TNM stage and lymphatic metastasis (all P < 0.05). The cell proliferation activity, cell migration rate, and the number of invasions of sh XIST group were lower than those of group C and NC (P < 0.05). The comparison of Ishikawa cell proliferation activity among C group, NC group, and sh XIST group at different time points showed: (1) The proliferation activity of Ishikawa cells at different time points was different (P < 0.05); (2) The proliferation activity of Ishikawa cells in the three groups was different (P < 0.05); (3) The change trend of Ishikawa cell proliferation activity in the three groups was different (P < 0.05). The double luciferase reporter gene showed that lncRNA XIST could target miR-101-3p.Conclusion lncRNA XIST and miR-101-3p are highly expressed in human endometrial carcinoma，and it is related to TNM stage and lymphatic metastasis. Inhibiting lncRNA XIST in Ishikawa cells can inhibit the malignant cell biological behavior of cancer cells, and its mechanism may be related to the targeted regulation of miR-101-3p.