尿道癌相关基因1通过微小RNA-513a-3p/细胞色素P_(450)1B1生物学轴促进人胃癌细胞增殖和迁移的机制研究

Mechanisms of urothelial carcinoma antigen 1 promoting the proliferation and migration of human gastric cancer cells through the micro RNA-513a-3p/cytochrome P_(450)1B1 axis

目的研究长链非编码RNA(lncRNA)尿路上皮癌胚抗原1(UCA1)在人胃癌细胞中的表达,并探讨UCA1通过微小RNA-513a-3p(miR-513a-3p)/细胞色素P_(450)1B1(CYP1B1)生物学轴对胃癌细胞增殖和迁移的影响及机制。方法分别向人胃癌细胞中转入UCA1小干扰RNA(siRNA)、miR-513a-3p模拟物和miR-513a-3p抑制剂以抑制UCA1、上调miR-513a-3p和抑制miR-513a-3p表达。通过定量PCR检测不同细胞UCA1、miR-513a-3p和CYP1B1 mRNA的表达,CCK8试剂盒检测细胞增殖,Transwell小室检测细胞迁移。结果与GES-1细胞相比,UCA1在人胃癌细胞HGC-27、SNU-5、AGS、SGC-7901中表达均升高(t=11.106,P<0.001;t=10.872,P<0.001;t=12.134,P<0.001;t=16.178,P<0.001),细胞增殖活性(t=7.252,P<0.001;t=8.964,P<0.001;t=8.666,P=0.003;t=8.697,P<0.001)和细胞迁移能力(t=4.040,P=0.010;t=7.079,P<0.001;t=6.833,P=0.003;t=7.527,P<0.001)均升高。双荧素酶基因报告实验显示miR-513a-3p与UCA1和CYP1B1靶向结合。与NC siRNA组相比,UCA1 siRNA显著下调UCA1和上调miR-513a-3p在SGC-7901细胞中的表达(t=7.895,P<0.001;t=12.911,P<0.001);miR-513a-3p表达上调显著抑制SGC-7901细胞中UCA1和CYP1B1 mRNA表达(t=7.224,P<0.001;t=5.645,P=0.002),而抑制miR-513a-3p则显著上调SGC-7901细胞中UCA1和CYP1B1 mRNA表达(t=9.628,P<0.001;t=11.665,P<0.001)。与单独UCA1敲低的SGC-7901细胞相比,UCA1和miR-513a-3p共同敲低的SGC-7901细胞CYP1B1 mRNA表达、细胞增殖活性和细胞迁移能力均升高(t=3.695,P=0.014;t=4.198,P=0.009;t=3.602,P=0.016)。结论UCA1通过miR-513a-3p/CYP1B1生物学轴促进人胃癌细胞的增殖和迁移。

Objective To study the expression levels of long non-coding RNA(lncRNA)urothelial carcinoma antigen 1(UCA1)in the human gastric cancer cells,and to explore the effect of promoting the proliferation and migration of human gastric cancer cells through the microRNA-513a-3p(miRNA-513a-3p)/cytochrome P_(450)1B1(CYP1B1)axis.Methods UCA1 small interference RNA(siRNA),miR-513a-3p-mimic and miR-513a-3p-inhibitor were introduced to human gastric cancer cells to inhibit UCA1,upregulate miR-513a-3p and inhibit miR-513a-3p expression.The expression of UCA1,miR-513a-3p and CYP1B1 mRNA was detected by quantitative PCR,cell proliferation was detected by the CCK8 kit,and cell migration was detected in Transwell chambers.Results Compared with GES-1 cells,UCA1 expression was significantly increased in the different gastric cancer cells(HGC-27,SNU-5,AGS,SGC-7901)(t=11.106,P<0.001;t=10.872,P<0.001;t=12.134,P<0.001;t=16.178,P<0.001),proliferation(t=7.252,P<0.001;t=8.964,P<0.001;t=8.666,P=0.003;t=8.697,P<0.001)and migration ability(t=4.040,P=0.010;t=7.079,P<0.001;t=6.833,P=0.003;t=7.527,P<0.001)were significantly increased.Dual fluorase gene reporting experiments showed that miR-513a-3p was targeted to UCA1 and CYP1B1.Compared with the NC siRNA group,UCA1 siRNA significantly down-regulated the expression of UCA1 and up-regulated the expression of miR-513a-3p in the SGC-7901 cells(t=7.895,P<0.001;t=12.911,P<0.001);the up-regulated expression of miR-513a-3p significantly inhibited the expression levels of UCA1 and CYP1B1 mRNA in the SGC-7901 cells(t=7.224,P<0.001;t=5.645,P=0.002),while inhibition of miR-513a-3p significantly up-regulated the expression levels of UCA1 and CYP1B1 mRNA in the SGC-7901 cells(t=9.628,P<0.001;t=11.665,P<0.001).Compared with the SGC-7901 cells with UCA1 knockdown alone,the CYP1B1 mRNA expression level,cell proliferation activity and cell migration ability of the SGC-7901 cells with UCA1 and miR-513a-3p knockdown were significantly increased(t=3.695,P=0.014;t=4.198,P=0.009;t=3.602,P=0.016).Conclusion UCA1 promotes the proliferation and migration of human gastric cancer cells through the miR-513a-3p/CYP1B1 biological axis.