摘要
目的探究长链非编码RNA(LncRNA)肿瘤蛋白P53靶基因1(TP53TG1)/微小RNA-96(miR-96)/原肌球蛋白1(TPM1)对胃癌细胞5-氟尿嘧啶(5-FU)耐药性的影响及其可能机制。方法体外培养SGC7901、SGC7901/5-FU细胞,将SGC7901/5-FU细胞分为对照组、TP53TG1 NC组(转染TP53TG1 NC)、TP53TG1 mimic组(转染TP53TG1 mimic)、miR-96 NC组(转染TP53TG1 mimic、miR-96 NC)和miR-96 mimic组(转染TP53TG1 mimic、miR-96 mimic)。UALCAN数据库分析胃腺癌组织、正常胃组织中TP53TG1表达差异;qRT-PCR法检测LncRNA TP53TG1、miR-96、TPM1 mRNA表达;CCK-8法、流式细胞术、蛋白质印迹法分别检测细胞增殖、凋亡及TPM1、Bcl-2、Bax蛋白表达;双荧光素酶实验检测miR-96、TPM1的靶向关系。结果胃腺癌组织中TP53TG1水平显著低于正常胃组织,差异有统计学意义(P<0.05)。与SGC7901细胞相比,SGC7901/5-FU细胞中LncRNA TP53TG1表达显著降低,miR-96表达显著升高,差异均有统计学意义(P<0.05)。与TP53TG1 NC组相比,TP53TG1 mimic组SGC7901/5-FU细胞中LncRNA TP53TG1、TPM1 mRNA、TPM1蛋白、Bax蛋白水平及细胞凋亡率显著升高,miR-96、Bcl-2蛋白水平及细胞吸光度(A)值显著降低,差异均有统计学意义(P<0.05);与miR-96 NC组相比,miR-96 mimic组SGC7901/5-FU细胞中LncRNA TP53TG1、TPM1 mRNA、TPM1蛋白、Bax蛋白水平及细胞凋亡率显著降低,miR-96、Bcl-2蛋白水平及细胞A值显著升高,差异均有统计学意义(P<0.05)。miR-96与TPM1在SGC7901/5-FU细胞中存在靶向关系。结论LncRNA TP53TG1可通过miR-96/TPM1轴降低胃癌细胞对5-FU的耐药性。
Objective To investigate the influence of long non-coding RNA(LncRNA)tumor protein p53 target gene 1(TP53TG1)/microRNA-96(miR-96)/tropomyosin 1(TPM1)on 5-fluorouracil(5-FU)resistance in gastric cancer cells and its possible mechanism.Methods SGC7901 and SGC7901/5-FU cells were cultured in vitro.SGC7901/5-FU cells were divided into control group,TP53TG1 NC group(transfected with TP53TG1 NC),TP53TG1 mimic group(transfected with TP53TG1 mimic),miR-96 NC group(transfected with TP53TG1 mimic and miR-96 NC)and miR-96 mimic group(transfected with TP53TG1 mimic and miR-96 mimic).UALCAN database analyze TP53TG1 expression in gastric adenocarcinoma and normal gastric tissues;qRT-PCR method were used to detect LncRNA TP53TG1,miR-96 and TPM1 expression;CCK-8,flow cytometry and WB were used to detect the cells proliferation,apoptosis and TPM1,Bcl-2,Bax protein expression,respectively;double luciferase assay were used to detect the targeting relationship between miR-96 and TPM1.Results TP53TG1 level in gastric adenocarcinoma was lower than that in normal gastric tissue,the difference was statistically significant(P<0.05).Compared with SGC7901 cells,TP53TG1 level in SGC7901/5-FU cells was lower,miR-96 level was significantly higher,the differences were statistically significant(P<0.05).Compared with TP53TG1 NC group,LncRNA TP53TG1,TPM1 mRNA,TPM1 protein,Bax protein levels and apoptosis rate of SGC7901/5-FU cells in TP53TG1 mimic group were higher,while miR-96,Bcl-2 protein levels and absorbance(A)value were lower,the differences were statistically significant(P<0.05);compared with miR-96 NC group,LncRNA TP53TG1,TPM1 mRNA,TPM1 protein,Bax protein levels and apoptosis rate of SGC7901/5-FU cells in miR-96 mimic group were lower,miR-96,Bcl-2 protein levels and A value were higher,the differences were statistically significant(P<0.05).There was a targeting relationship between miR-96 and TPM1 in SGC7901/5-FU cells.Conclusion LncRNA TP53TG1 can reduce the resistance of gastric cancer cells to 5-FU through miR-96/TPM1 axis.
作者
文俏程
卢善政
张蔚林
肖志刚
WEN Qiao-cheng;LU Shan-zheng;ZHANG Wei-lin(Department of General Surgery,Hunan Provincial People's Hospital(the First Affiliated Hospital of Hunan Normal University),Changsha Hunan 410005,China)
出处
《临床和实验医学杂志》
2021年第23期2492-2496,共5页
Journal of Clinical and Experimental Medicine
基金
湖南省自然科学基金项目(编号:2020JJ5305)。
