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中国癌症防治杂志

中国癌症防治杂志 ›› 2019, Vol. 11 ›› Issue (5): 375-381.doi: 10.3969/j.issn.1674-5671.2019.05.03

• 基础研究 • 上一篇    下一篇

IncRNA MAFG-AS1及miR-143-3p对宫颈癌细胞增殖、凋亡的影响及机制研究

  

  1. 郴州市第一人民医院妇科，产科
  • 出版日期:2019-10-25 发布日期:2019-11-25

Effect of lncRNA MAFG-AS1 and miR-143-3p on the proliferation and apoptosis of cervical cancer cells and its mechanism

  • Online:2019-10-25 Published:2019-11-25

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摘要： 目的 探讨lncRNA MAFG-AS1和miR-143-3p对宫颈癌细胞增殖和凋亡的影响及其潜在的作用机制。方法 MAFG-AS1和miR-143-3p表达载体转染至宫颈癌细胞Hela后，采用qRT-PCR检测宫颈癌组织及相应癌旁正常组织，宫颈癌细胞Hela和正常宫颈细胞株Ect1/E6E7中miR-143-3p和MAFG-AS1 mRNA的表达水平；Western blot检测增殖相关蛋白Bcl-2、Cyclin D1及凋亡相关蛋白Bax、cleaved-Caspase-3、p21、p27的表达；MTT法检测细胞增殖活性；流式细胞术检测细胞凋亡；双荧光素酶报告基因突变检测lncRNA MAFG-AS1与miR-143-3p的相互作用。结果 与癌旁正常组织相比，宫颈癌组织中MAFG-AS1 mRNA表达升高（0.905±0.115 vs 2.835±0.164，t=43.091，P<0.001），miR-143-3p表达降低（0.944±0.075 vs 0.382±0.071，t=24.336，P<0.001）；相较于正常宫颈细胞株Ect1/E6E7，宫颈癌细胞Hela中MAFG-AS1 mRNA的表达显著升高（P<0.001），miR-143-3p表达显著降低（P<0.001）。下调MAFG-AS1和过表达miR-143-3p均可抑制Hela细胞增殖活性，促进细胞凋亡；抑制Bcl-2和Cyclin D1蛋白表达，促进Bax、Caspase-3、p21、p27蛋白表达。MAFG-AS1可靶向调控miR-143-3p表达，且抑制miR-143-3p表达可逆转下调MAFG-AS1表达对Hela细胞抑制细胞增殖、促进细胞凋亡作用。结论 lncRNA MAFG-AS1可抑制宫颈癌细胞增殖，促进凋亡，其机制可能与靶向调控miR-143-3p有关。

关键词: 宫颈癌, lncRNA MAFG-AS1, miR-143-3p, 增殖, 凋亡

Abstract:

Objective To investigate the effects of lncRNA MAFG-AS1 and miR-143-3p on proliferation and apoptosis of cervical cancer cells and its potential mechanism. Methods  After MAFG-AS1 and miR-143-3p expression vectors were transfected into Hela cells，qRT-PCR was used to detect the expression of miR-143-3p and MAFG-AS1 mRNA in Hela cells； protein expression，cell proli-feration activity and apoptosis were detected by Western blot， MTT assay and flow cytometry， respectively；dual-luciferase reporter gene was used to the interaction between lncRNA MAFG-AS1 and miR-143-3p. Results Compared with adjacent normal tissues，MAFG-AS1 mRNA expression was increased in cervical cancer tissues （0.905±0.115 vs 2.835±0.164，t=43.091，P<0.001），and miR-143-3p expression level was decreased （0.944±0.075 vs  0.382±0.071，t=24.336，P<0.001）. Compared with the normal cervical cell line Ect1/E6E7，the expression of MAFG-AS1 mRNA in Hela cells was significantly increased（P<0.001），and the expression of miR-143-3p was significantly decreased（P<0.001）. Down-regulation of MAFG-AS1 and miR-143-3p overexpression inhibited the proliferation of Hela cells and promoted apoptosis， inhibited the expression of Bcl-2 and Cyclin D1 proteins，and promoted the expression of Bax，Caspase-3，p21，and p27 proteins. MAFG-AS1 targeted to regulate the expression of miR-143-3p. Inhibition of miR-143-3p expression reversed the effect of MAFG-AS1 on inhibiting proliferation and promoting apoptosis Hela cells. Conclusions lncRNA MAFG-AS1 can inhibit the proliferation of cervical cancer cells and promote their apoptosis. The mechanism may be related to miR-143-3p，which will provide new targets for the prevention and treatment of cervical cancer.

Key words: Cervical cancer, lncRNA MAFG-AS1, miR-143-3p, Proliferation, Apoptosis

中图分类号: 

  • R737.33