摘要
目的:研究泛素结合酶9(Ubc9)通过核因子κB(NF-κB)通路诱导胃癌细胞SGC7901凋亡的影响。方法:采用Ubc9基因的质粒对胃癌SGC7901细胞株进行瞬时转染,将实验对象分为:正常胃癌细胞A组(Normal组),脂质体B组(Lipo-2000组),空载质粒C组(NC-shRNA组),及目的质粒D组(Ubc9-shRNA组)。采用qRT-PCR检测各组Ubc9 mRNA水平,Western blot检测各组Ubc9、α-SMA、IKBα、p65与Bcl-2蛋白表达结果,流式细胞仪检测各组细胞周期与凋亡结果。结果:与各组比较,Ubc9-shRNA组转染后细胞凋亡增加,差异有统计学意义(P<0.05);Ubc9-shRNA组能显著的抑制胃癌SGC7901中Ubc9基因的表达,差异有统计学意义(P<0.05);Ubc9-shRNA组明显抑制SGC7901细胞中Ubc9蛋白、α-SMA和p65、Bcl-2蛋白表达,促进IKBα表达量的增加,差异有统计学意义(P<0.05)。结论:Ubc9可能通过影响NF-κB通路下游凋亡基因从而参与胃癌细胞的凋亡。
Objective:To study the effect of Ubc9 on apoptosis of SGC7901 in gastric cancer cells by NF-κB pathway.Methods:Gastric cancer SGC7901 cell line was transfected by Ubc9 gene plasmid.The experimental subjects were divided into Normal gastric cancer cell group A (Normal group),liposome group B (LiPO-2000 group),no-load plasmid group C (NC-shRNA group),and target plasmid group D (Ubc9-shRNA group).The mRNA levels of Ubc9 in each group were detected by qRT-PCR,and the protein expressions of Ubc9,α-SMA,IKBα,p65 and Bcl-2 in each group were detected by Western blot.Cell cycle and apoptosis were detected by flow cytometry.Results:Compared with each group,apoptosis increased after transfection in Ubc9-shRNA group,and the difference was statistically significant (P<0.05).Ubc9-shRNA group could significantly inhibit the expression of Ubc9 gene in SGC7901 of gastric cancer,with statistical significance (P<0.05);Ubc9-shRNA group significantly inhibited the expression of Ubc9 protein,α-SMA,p65,Bcl-2 protein in SGC7901 cells,and promoted the expression of IKBα,with statistical significance (P<0.05).Conclusion:Ubc9 may participate in the apoptosis of gastric cancer cells by affecting the downstream apoptotic genes of NF-κB pathway.
作者
袁金华
曾宪晶
刘伍才
罗亮
YUAN Jinhua;ZENG Xianjing;LIU Wucai;LUO Liang(Affiliated Hospital of Jinggangshan University,Ji'an Jiangxi 343000,China)
出处
《药品评价》
CAS
2022年第3期145-148,165,共5页
Drug Evaluation
基金
江西省卫健委科技计划项目(2020-3792号)。
关键词
胃肿瘤
核因子-ΚB
泛素结合酶9
凋亡基因
Stomach neoplasms
Nuclear factor-κB
Ubiquitin binding enzyme 9(Ubc9)
Apoptosis gene
