Hsa-miR-5195-3p对人宫颈癌细胞SiHa增殖、迁移与侵袭的影响 <sup>*</sup>

doi:10.13523/j.cb.1909030

中国生物工程杂志

2020, Vol. 40

Issue (4): 17-24 DOI: 10.13523/j.cb.1909030

研究报告

Hsa-miR-5195-3p对人宫颈癌细胞SiHa增殖、迁移与侵袭的影响 ^*

唐敏,万群,孙恃雷,胡静,孙子久,方玉婷,张彦(

)

重庆医科大学检验医学院临床检验诊断学教育部重点实验室重庆 400016

The Effects of Hsa-miR-5195-3p on the Proliferation, Migration and Invasion of Human Cervical Cancer SiHa Cells

TANG Min,WAN Qun,SUN Shi-lei,HU Jing,SUN Zi-jiu,FANG Yu-ting,ZHANG Yan(

)

Key Laboratory of Diagnostic Medicine Designated by the Chinese Ministry of Education, Chongqing Medical University, Chongqing 400016, China

全文: PDF(1144 KB) HTML

摘要：

目的：探讨miR-5195-3p对人宫颈癌细胞系SiHa增殖、迁移与侵袭的影响。方法：采用qRT-PCR检测人宫颈癌细胞SiHa和正常上皮细胞HaCaT中miR-5195-3p的表达水平。将miR-5195-3p mimic转染至SiHa细胞中构建外源性过表达细胞株,阴性对照组中则转染NC mimic,并用qRT-PCR验证转染效率;通过MTT和集落形成实验检测细胞增殖能力;划痕愈合实验检测细胞横向迁移能力;Transwell小室实验检测细胞纵向迁移能力和侵袭能力;采用qRT-PCR和Western blot检测E-cadherin、Vimentin与snail mRNA转录水平及蛋白表达水平。结果：宫颈癌细胞SiHa中的miR-5195-3p表达水平较HaCaT偏低(P<0.05)。与阴性对照组相比,转染miR-5195-3p mimic的SiHa细胞中miR-5195-3p水平显著增高(P<0.01);并且其体外增殖(P<0.001),迁移(P<0.001)与侵袭能力(P<0.001)明显减弱;同时E-cadherin表达水平上调而Vimentin、snail表达水平下调。结论：过表达miR-5195-3p可能通过阻碍EMT通路抑制宫颈癌细胞SiHa的增殖,迁移与侵袭。

关键词： 宫颈癌; miR-5195-3p; 增殖; 迁移; 侵袭

Abstract:

Objective: To investigate the effect of miR-5195-3p on the proliferation, migration and invasion of human cervical cancer cell line SiHa. Methods: The expression levels of miR-5195-3p in human cervical cancer cells SiHa and normal epithelial cells HaCaT were detected by real-time quantitative PCR.The miR-5195-3p mimic was transfected into SiHa cells to construct exogenous overexpressed cell lines,while the negative control group was transfected with NC mimic and the transfection efficiency was verified by real-time quantitative PCR;MTT and colony formation experiments were utilized to assess the proliferation capacity; The competence of transversal migration was examined by Wound healing experiment;Transwell assay was performed to evaluated the longitudinal migration and invasion ability;The mRNA transcription and protein expression levels of E-cadherin, Vimentin and snail were analyzed by real-time quantitative PCR and Western blot. Results: the expression level of miR-5195-3p in SiHa was lower than HaCaT (P<0.05).Compared with the negative control group, the level of miR-5195-3p in SiHa cell which transfected the miR-5195-3p mimic was significantly increased (P<0.01);Moreover, their proliferation,migration and invasion in vitro were distinctly reduced(P<0.001);Meanwhile, the expression levels of E-cadherin were up-regulated while those of Vimentin and snail were down-regulated. Conclusion: overexpression of miR-5195-3p may inhibit the proliferation, migration and invasion of cervical cancer cell SiHa by blocking EMT pathway.

Key words: Cervical Cancer miR-5195-3p Proliferation Migration Invasion

收稿日期: 2019-09-16 出版日期: 2020-05-18

ZTFLH:

Q291

基金资助: * 国家自然科学基金项目(81172017);国家自然科学基金项目(81974449)

通讯作者: 张彦 E-mail: zy2753@hotmail.com

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引用本文:

唐敏,万群,孙恃雷,胡静,孙子久,方玉婷,张彦. Hsa-miR-5195-3p对人宫颈癌细胞SiHa增殖、迁移与侵袭的影响 ^*[J]. 中国生物工程杂志, 2020, 40(4): 17-24.

TANG Min,WAN Qun,SUN Shi-lei,HU Jing,SUN Zi-jiu,FANG Yu-ting,ZHANG Yan. The Effects of Hsa-miR-5195-3p on the Proliferation, Migration and Invasion of Human Cervical Cancer SiHa Cells. China Biotechnology, 2020, 40(4): 17-24.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.1909030 或 https://manu60.magtech.com.cn/biotech/CN/Y2020/V40/I4/17

表1 引物序列

图1 miR-5195-3p在正常上皮细胞HaCaT与宫颈癌细胞SiHa中的表达

图2 miR-5195-3p促进宫颈癌细胞SiHa的增殖

图3 miR-5195-3p促进宫颈癌细胞SiHa的迁移

图4 miR-5195-3p促进宫颈癌细胞SiHa的侵袭

图5 miR-5195-3p激活SiHa细胞的EMT信号通路

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