lncRNA GAS5对宫颈癌细胞增殖、侵袭能力及PTEN/Akt/mTOR信号通路的影响

doi:10.11904/j.issn.1002-3070.2024.02.004

摘要/Abstract

摘要： 目的探讨长链非编码RNA GAS5(lncRNA GAS5)对宫颈癌细胞增殖、侵袭能力及PTEN/Akt/mTOR信号通路的影响,阐明lncRNA GAS5对宫颈癌可能的作用机制。方法实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测人宫颈癌细胞HeLa、C33A、Caski、Siha和人正常子宫颈上皮细胞End1/E6E7中lncRNA GAS5表达水平。将HeLa细胞分为空白对照组(BC组)、阴性对照组(NC组)和过表达lncRNA GAS5组(GAS5组);将Siha细胞分为空白对照组(BC组)、阴性对照siRNA组(si-NC组)和siRNA-lncRNA GAS5组(si-GAS5)。CCK-8法测定细胞增殖,Transwell小室测定细胞侵袭和迁移能力,Western blot测定细胞中PCNA、MMP-2、MMP-9、PTEN、Akt、p-Akt、mTOR、p-mTOR蛋白表达水平。结果宫颈癌细胞HeLa、C33A、Caski、Siha中lncRNA GAS5水平均低于正常宫颈上皮细胞End1/E6E7(P＜0.001)。与NC组比较,HeLa细胞过表达lncRNA GAS5后,细胞增殖、侵袭和迁移能力以及PCNA、MMP-2、MMP-9、p-Akt和p-mTOR蛋白水平降低(P＜0.001),PTEN蛋白水平升高(P＜0.001)。与si-NC组比较,在Siha细胞中抑制lncRNA GAS5表达后,细胞增殖、侵袭和迁移能力以及PCNA、MMP-2、MMP-9、p-Akt和p-mTOR蛋白水平升高(P＜0.001),PTEN蛋白水平降低(P＜0.001)。结论宫颈癌细胞lncRNA GAS5水平降低,lncRNA GAS5可抑制宫颈癌细胞增殖、侵袭迁移能力和PTEN/Akt/mTOR信号通路激活。

关键词: 长链非编码RNA, 宫颈癌, 细胞

Abstract: Objective This Objective of this study was to investigate the effects of long chain non-coding RNA GAS5(lncRNA GAS5)on the proliferation,invasion ability and PTEN/Akt/mTOR signaling pathway of cervical cancer cells,and to elucidate the possible mechanism of lncRNA GAS5 in cervical cancer. Methods Real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to detect the levels of lncRNA GAS5 in human cervical cancer HeLa cells,C33A cells,Caski cells,Siha cells and normal cervical epithelial End1/E6E7 cells.HeLa cells were divided into the blank control group(BC group),negative control group(NC group)and overexpressing lncRNA GAS5 group(GAS5 group);Siha cells were divided into the blank control group(BC group),negative control siRNA group(si-NC group)and siRNA-lncRNA GAS5 group(si-GAS5).The CCK8 assay was used to measure proliferation of cervical cancer cells.The Transwell was used to measure the invasion and migration capabilities of cervical cancer cells.Western blot was used to determine the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase(MMP)-2,MMP-9,PTEN,Akt,phosphorylated Akt(p-Akt),mTOR and p-mTOR proteins in cervical cancer cells. Results The results showed that the levels of lncRNA GAS5 in HeLa cells,C33A cells,Caski cells and Siha cells were lower than that in End1/E6E7 cells(P＜0.001).Compared with the NC group,overexpression of lncRNA GAS5 in HeLa cells resulted in a decrease in the cell proliferation,invasion,and migration abilities,as well as a decrease in the levels of PCNA,MMP-2,MMP-9,p-Akt,and p-mTOR proteins(P＜0.001),while an increase in the level of PTEN protein(P＜0.001).Compared with the si-NC group,inhibition of lncRNA GAS5 expression in Siha cells resulted in increased the cell proliferation,invasion,and migration abilities,as well as increased the levels of PCNA,MMP-2,MMP-9,p-Akt,and p-mTOR proteins(P＜0.001),while decreased the level of PTEN protein(P＜0.001). Conclusion The level of lncRNA GAS5 in cervical cancer cells is reduced,and lncRNA GAS5 can inhibit the proliferation,invasion and migration abilities of cervical cancer cells,as well as activate the PTEN/Akt/mTOR signaling pathway.

Key words: lncRNA, Cervical cancer, cells

中图分类号:

R737.33

周建云, 储巧香, 陆月梅, 朱静, 丁其培. lncRNA GAS5对宫颈癌细胞增殖、侵袭能力及PTEN/Akt/mTOR信号通路的影响[J]. 实用肿瘤学杂志, 2024, 38(2): 96-103.

ZHOU Jianyun, CHU Qiaoxiang, LU Yuemei, ZHU Jing, DING Qipei. The effects of lncRNA GAS5 on the proliferation,invasion ability and PTEN/Akt/mTOR signaling pathway of cervical cancer cells[J]. Journal of Practical Oncology, 2024, 38(2): 96-103.

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