LncRNA MYLK-AS1调节miR-141-3p/STMN1轴对胃癌细胞增殖、凋亡和侵袭的影响

刘洁, 谢兴明, 钮洪霞, 杨先智

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重庆医科大学学报 ›› 2024, Vol. 49 ›› Issue (03) : 276-282. DOI: 10.13406/j.cnki.cyxb.003458
基础研究

LncRNA MYLK-AS1调节miR-141-3p/STMN1轴对胃癌细胞增殖、凋亡和侵袭的影响

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Impacts of LncRNA MYLK-AS1 on proliferation,apoptosis,and invasion of gastric cancer cells by regulating the miR-141-3p/STMN1 axis

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摘要

目的 探讨长链非编码RNA(long non-coding RNA,LncRNA)肌球蛋白轻链激酶反义RNA1(myosin light chain kinase antisense RNA1,MYLK-AS1)调节miR-141-3p/微管不稳定蛋白1(stathmin 1,STMN1)轴对胃癌细胞增殖、凋亡和侵袭的影响。 方法 将HGC27细胞分为NC组、si-NC组、si-MYLK-AS1组、si-MYLK-AS1+inhibitor NC组、si-MYLK-AS1+miR-141-3p inhibitor组。双萤光素酶报告基因实验检测LncRNA MYLK-AS1、miR-141-3p、STMN1的关系;qRT-PCR检测HGC27细胞中LncRNA MYLK-AS1、miR-141-3p表达;CCK-8法检测HGC27细胞增殖情况;流式细胞术检测HGC27细胞凋亡;使用Transwell实验评估了HGC27细胞的侵袭和迁移能力,并统计了穿透基质膜的细胞数量;同时采用Western blot技术检测了HGC27细胞中STMN1、E-cadherin、Vimentin及N-cadherin这几种蛋白表达量的变化。 结果 HGC27细胞中LncRNA MYLK-AS1、STMN1水平高于GES-1细胞(P<0.05),miR-141-3p水平低于GES-1细胞(P<0.05)。si-MYLK-AS1组HGC27细胞A450 nm值、迁移、侵袭细胞数量、LncRNA MYLK-AS1表达量、STMN1、N-cadherin、Vimentin蛋白水平低于NC组、si-NC组(P<0.05),HGC27细胞凋亡率、miR-141-3p表达量、E-cadherin蛋白水平高于NC组、si-NC组(P<0.05);而miR-141-3p低表达减弱了沉默LncRNA MYLK-AS1抑制HGC27细胞发展的作用;LncRNA MYLK-AS1靶向调节miR-141-3p/STMN1轴。 结论 LncRNA MYLK-AS1可能通过上调microRNA-141-3p的表达水平,间接导致STMN1基因表达受到抑制,这一过程可能对胃癌细胞的增殖、凋亡及侵袭特性产生明显影响。

Abstract

Objective To investigate the impacts of long non-coding RNA(LncRNA) myosin light chain kinase antisense RNA1 (MYLK-AS1) on proliferation,apoptosis,and invasion of gastric cancer cells by regulating the miR-141-3p/stathmin 1(STMN1) axis. Methods HGC27 cells were grouped into negative control(NC) group,si-NC group,si-MYLK-AS1 group,si-MYLK-AS1+inhibitor NC group,and si-MYLK-AS1+miR-141-3p inhibitor group. The relationship among LncRNA MYLK-AS1,miR-141-3p,and STMN1 was determined by the dual-luciferase reporter gene assay;the expression of LncRNA MYLK-AS1 and miR-141-3p in HGC27 cells was measured by qRT-PCR;the CCK-8 method and flow cytometry were usde to determined the proliferation and apoptosis of HGC27 cells,respectively;the numbers of invading and migrating HGC27 cells were counted by Transwell;the protein levels of STMN1,E-cadherin,Vimentin,and N-cadherin in HGC27 cells were measured by Western blot. Results Compared with GES-1 cells,the levels of LncRNA MYLK-AS1 and STMN1 in HGC27 cells were significantly up-regulated(P<0.05),and the level of miR-141-3p was significantly down-regulated(P<0.05). Compared with the NC group and si-NC group,the si-MYLK-AS1 group showed significant decreases in the optical density at 450 nm,numbers of migrating and invading cells,expression level of LncRNA MYLK-AS1,and protein levels of STMN1,N-cadherin,and Vimentin(P<0.05),as well as significant increases in the apoptosis rate,expression level of miR-141-3p,and protein level of E-cadherin(P<0.05). However,the low expression of miR-141-3p attenuated the inhibition of HGC27 cell development by silencing LncRNA MYLK-AS1;LncRNA MYLK-AS1 regulated the miR-141-3p/STMN1 axis in a targeted manner. Conclusion Silencing LncRNA MYLK-AS1 may inhibit the expression of STMN1 by up-regulating miR-141-3p,thus affecting the proliferation,apoptosis,and invasion of gastric cancer cells.

关键词

长链非编码RNA肌球蛋白轻链激酶反义RNA1 / 微小RNA-141-3p/微管不稳定蛋白1轴 / 胃癌 / 增殖 / 凋亡 / 侵袭

Key words

long non-coding RNA myosin light chain kinase antisense RNA1 / miR-141-3p/stathmin 1 axis / gastric cancer / proliferation / apoptosis / invasion

中图分类号

R735.2

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刘洁 , 谢兴明 , 钮洪霞 , . LncRNA MYLK-AS1调节miR-141-3p/STMN1轴对胃癌细胞增殖、凋亡和侵袭的影响. 重庆医科大学学报. 2024, 49(03): 276-282 https://doi.org/10.13406/j.cnki.cyxb.003458
Liu Jie, Xie Xingming, Niu Hongxia, et al. Impacts of LncRNA MYLK-AS1 on proliferation,apoptosis,and invasion of gastric cancer cells by regulating the miR-141-3p/STMN1 axis[J]. Journal of Chongqing Medical University. 2024, 49(03): 276-282 https://doi.org/10.13406/j.cnki.cyxb.003458

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基金

遵义市科技局科技计划资助项目(遵市科合HZ字[2019]159)

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