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

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

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A_{450 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

基金资助

遵义市科技局科技计划资助项目(遵市科合HZ字[2019]159)