LncRNA EIF3J-AS1通过靶向调控miR-597-5p对胃癌细胞增殖、周期分布和凋亡的实验研究

LncRNA EIF3J-AS1 regulates cell proliferation,cycle distribution and apoptosis in gastric cancer via targeting miR-597-5p

目的探讨LncRNA EIF3J-AS1靶向miR-597-5p对胃癌细胞增殖、周期分布和凋亡的影响。方法RT-qPCR分析LncRNA EIF3J-AS1和miR-597-5p在胃癌组织、癌旁组织、胃癌细胞(SNU-1、AGS、HS-746T)以及人胃黏膜细胞GES1中的表达。将si-LncRNA EIF3J-AS1、si-NC、miR-NC、miR-597-5p mimics、pcDNA、pcDNA-LncRNA EIF3J-AS1、si-LncRNA EIF3J-AS1与anti-miR-NC、si-LncRNA EIF3J-AS1与anti-miR-597-5p分别转染SNU-1细胞,流式细胞术检测细胞周期分布和凋亡率。CCK-8法检测细胞活力。LncRNA EIF3J-AS1和miR-597-5p的靶向关系通过双荧光素酶报告实验来证实。结果胃癌组织中LncRNA EIF3J-AS1表达水平显著高于癌旁组织(P<0.05),miR-597-5p表达水平显著低于癌旁组织(P<0.05)。胃癌细胞中LncRNA EIF3J-AS1表达水平显著高于GES1细胞(P<0.05),miR-597-5p表达水平显著低于GES1细胞(P<0.05)。沉默LncRNA EIF3J-AS1表达显著降低细胞活力、S期细胞比例(P<0.05),增加凋亡率和G0/G1期细胞比例(P<0.05),上调miR-597-5p表达水平(P<0.05)。过表达miR-597-5p显著降低细胞活力、S期细胞比例(P<0.05),增加凋亡率和G0/G1期细胞比例(P<0.05)。过表达LncRNA EIF3J-AS1显著降低miR-597-5p表达水平(P<0.05)。LncRNA EIF3J-AS1与miR-597-5p直接特异性结合。抑制miR-597-5p表达显著削弱沉默LncRNA EIF3J-AS1对SNU-1细胞活力、周期分布和凋亡的影响(P<0.05)。结论沉默LncRNA EIF3J-AS1通过靶向上调miR-597-5p可抑制胃癌细胞增殖,阻碍周期进展,并诱导细胞凋亡。

Objective To investigate the effects of LncRNA EIF3J-AS1 targeting miR-597-5p on the proliferation,cycle distribution and apoptosis of gastric cancer cells.Methods LncRNA EIF3J-AS1 and miR-597-5p expression in gastric cancer tissues,adjacent tissues,gastric cancer cells(SNU-1,AGS,HS-746T)and human gastric mucosal cells GES1 were calculated using RT-qPCR.si-LncRNA EIF3J-AS1,si-NC,miR-NC,miR-597-5p mimics,pcDNA,pcDNA-LncRNA EIF3J-AS1,si-LncRNA EIF3J-AS1 and anti-miR-NC,si-LncRNA EIF3J-AS1 and anti-miR-597-5p were transfected into SNU-1 cells,respectively.And the cell cycle distribution and apoptosis rate was assessed by flow cytometry.CCK-8 method was applied to detect cell viability.The targeting relationship between LncRNA EIF3J-AS1 and miR-597-5p was confirmed using dual luciferase reporter assay.Results The expression level of LncRNA EIF3J-AS1 in gastric cancer tissue was significantly higher than that in adjacent tissues(P<0.05),while the expression level of miR-597-5p was significantly lower than that in adjacent tissues(P<0.05).The expression level of LncRNA EIF3J-AS1 in gastric cancer cells was significantly higher than that of GES1 cells(P<0.05),while the expression level of miR-597-5p was significantly lower than that of GES1 cells(P<0.05).Silencing LncRNA EIF3J-AS1 expression significantly reduced cell viability,S phase cell ratio(P<0.05),increased apoptosis rate and G0/G1 phase cell ratio(P<0.05),as well as increased miR-597-5p expression level(P<0.05).Overexpression of miR-597-5p significantly reduced cell viability,S phase cell ratio(P<0.05),increased apoptosis rate and G0/G1 phase cell ratio(P<0.05).Overexpression of LncRNA EIF3J-AS1 significantly reduced the expression level of miR-597-5p(P<0.05).LncRNA EIF3J-AS1 specifically and directly binds to miR-597-5p.miR-597-5p inhibition significantly weakened the effect of silencing LncRNA EIF3J-AS1 on the viability,cycle distribution and apoptosis of SNU-1 cells(P<0.05).Conclusion Silencing LncRNA EIF3J-AS1 suppresses cell proliferation,blocks cell cycle progression and induces cell apoptosis in gastric cancer by targeting and up-regulating miR-597-5p.