miR-1-3p靶向TNKS2抑制宫颈癌细胞增殖、迁移和侵袭

miR-1-3p targets TNKS2 to inhibit proliferation,migration and invasion of cervical cancer cells

目的探讨miR-1-3p靶向端锚聚合酶2(TNKS2)基因对宫颈癌Ca-Ski细胞增殖、迁移及侵袭的调控作用。方法采用实时荧光定量聚合酶链式反应(q RT-PCR)检测正常宫颈鳞状细胞及不同宫颈癌细胞中miR-1-3p的表达情况,选取宫颈癌Ca-Ski细胞进行转染实验,利用脂质体转染法将miR-1-3p模拟物(miR-1-3p mimics)及模拟物对照(NC mimics)分别转染至Ca-Ski细胞,分别记为miR-1-3p mimics组和miR-NC组,将未经转染处理的Ca-Ski细胞记为Blank组(空白对照)。qRT-PCR检测各组Ca-Ski细胞中miR-1-3p的表达,噻唑蓝比色法(MTT)检测细胞增殖能力,Transwell实验检测细胞迁移和侵袭能力,通过生物信息学软件分析和双荧光素酶报告基因实验验证miR-1-3p和TNKS2靶向关系,蛋白免疫印迹法(Western blot)检测各组细胞中TNKS2蛋白表达。结果 miR-1-3p在宫颈癌细胞中的表达显著低于正常宫颈鳞状细胞(P<0.05)。转染miR-1-3p mimics 48 h后,Ca-Ski细胞中miR-1-3p的表达显著升高(P<0.05)。过表达miR-1-3p能够明显抑制Ca-Ski细胞的增殖、迁移和侵袭能力(P<0.05)。生物信息学软件分析显示miR-1-3p和TNKS2 3’UTR存在结合位点,双荧光素酶报告基因实验显示TNKS2是miR-1-3p的靶基因,Western blot检测结果显示miR-1-3p可抑制TNKS2的表达。结论 miR-1-3p可靶向TNKS2基因抑制宫颈癌Ca-Ski细胞增殖、迁移及侵袭。

Objective To investigate the regulation of miR-1-3 p targeting tankyrase 2(TNKS2)gene on proliferation,migration and invasion of cervical cancer Ca-Ski cells. Methods q RT-PCR was used to detect the expression of miR-1-3 p in normal cervical squamous cells and different cervical cancer cells. The cervical cancer Ca-Ski cells were selected for transfection experiments. The liposome transfection method was used to transfect miR-1-3 p mimics and NC mimics into CaSki cells,respectively as miR-1-3 p mimics group and miR-NC group;Ca-Ski cells without transfection treatment were used as Blank group. qRT-PCR was used to detect the expression of miR-1-3 p in Ca-Ski cells. MTT was used to detect cell proliferation. Transwell assay was used to detect cell migration and invasion. Bioinformatics software analysis and double luciferase reporter gene experiments were performed to verify the targeting relationship between miR-1-3 p and TNKS2.Western blot was used to detect the expression of TNKS2 protein in each group of cells. Results The expression of miR-1-3 p in cervical cancer cells was significantly lower than that in normal cervical squamous cells(P<0.05). After transfection of miR-1-3 p mimics for 48 h,the expression of miR-1-3 p was significantly increased in Ca-Ski cells(P<0.05).Overexpression of miR-1-3 p significantly inhibited the proliferation,migration and invasion of Ca-Ski cells(P<0.05).Bioinformatics software analysis showed that there was a binding site for miR-1-3 p and TNKS2 3′UTR. The dual luciferase reporter gene assay showed that TNKS2 was the target gene of miR-1-3 p,and the results of Western blot showed that miR-1-3 p could inhibit the expression of TNKS2. Conclusion miR-1-3 p could target TNKS2 gene to inhibit the proliferation,migration and invasion of cervical cancer Ca-Ski cells.