敲低长链非编码RNA LOXL1- AS1表达对胃癌细胞增殖、凋亡的影响及其机制

Effects of LOXL1-AS1 knockdown on proliferation and apoptosis of gastric cancer cells

目的探讨敲低长链非编码RNA LOXL1-AS1表达对胃癌细胞增殖、凋亡的影响,并探讨其可能的作用机制。方法实时荧光定量PCR(qRT-PCR)法检测不同胃癌细胞株(AGS、BGC823、NCI-N87、MGC803和SGC7901)和正常胃黏膜细胞株GES-1中LOXL1-AS1表达情况;选取LOXL1-AS1表达水平最高的胃癌BGC823细胞系,建立LOXL1-AS1敲低模型,实验分为sh-LOXL1-AS1组和sh-NC组,分别转染LOXL1-AS1靶向shRNA和scramble shRNA,qRT-PCR法验证干扰效率,CCK-8检测细胞增殖,流式细胞仪检测细胞凋亡情况,免疫印迹法检测Bcl-2、Bax、PI3K、p-PI3K、Akt和p-Akt表达。结果 qRT-PCR结果显示,与正常胃黏膜细胞GES-1相比,胃癌细胞株中LOXL1-AS1表达水平均显著升高(P<0.05)。与sh-NC组比较,sh-LOXL1-AS1组LOXL1-AS1表达水平显著降低(P<0.05),细胞增殖率显著降低(P<0.05),细胞凋亡率显著升高(P<0.05),细胞Bcl-2、p-PI3K和p-Akt表达显著降低,Bax表达显著升高(P<0.05)。结论敲低LOXL1-AS1表达可抑制胃癌细胞增殖并促进细胞凋亡,其机制可能与抑制PI3K/Akt信号通路,继而调控凋亡相关蛋白表达,启动细胞凋亡程序有关。

Objective To investigate the effects of LOXL1-AS1 knockdown on proliferation and apoptosis of gastric cancer cells, and explore the possible mechanisms. Methods Quantitive realtime PCR(qRT-PCR) was used to detect the expression of lncRNA LOXL1-AS1 in gastric cancer cell lines(AGS, BGC823, NCI-N87, MGC803 and SGC7901) and the normal gastric mucosal cell line GES-1. BGC823 cells were randomly divided into two groups: sh-NC group and sh-LOXL1-AS1 group. Scramble shRNA and LOXL1-AS1 shRNA were transfected into sh-NC group and sh-LOXL1-AS1 group, respectively. The interference efficiency of LOXL1-AS1 was detected by qRT-PCR. The cell proliferation was measured by CCK-8 assay. Apoptosis was detected by PI/Annexin V double staining flow cytometry. The expressions of Bax, Bcl-2, PI3 K, p-PI3 K, Akt and p-Akt were evaluated by Western blotting. Results Compared with normal gastric mucosal cell GES-1, the expression of LOXL1-AS1 in gastric cancer cell lines was increased(P<0.05). Compared with the sh-NC group, the expression of LOXL1-AS1 was decreased significantly in sh-LOXL1-AS1 group(P<0.05). Besides, the cell proliferation was decreased significantly in sh-LOXL1-AS1 group(P<0.05). The apoptosis rate was increased significantly in sh-LOXL1-AS1 group(P<0.05). Moreover, the expression of Bax was up-regulated, but the expressions of Blc-2, p-PI3 K and p-Akt were down-regulated in sh-LOXL1-AS1 group(P<0.05). Conclusion LOXL1-AS1 knockdown could inhibit the cell proliferation and promote the apoptosis of gastric cancer cells by inhibiting the PI3 K/Akt signaling pathway, thus regulating the expressions of apoptosis-related proteins.