长链非编码RNA HSALNT0000015对结直肠癌细胞生物学行为的影响

Effect of lncRNA HSALNT0000015 on the biological behavior of colorectal cancer cells

目的探讨长链非编码RNA(lncRNA)HSALNT0000015对结直肠癌细胞生物学行为的影响及其作用机制。方法采用实时荧光定量聚合酶链反应(qRT-PCR)检测80例结直肠癌组织及配对的15例癌旁组织,同时检测人结直肠癌细胞(Lovo、SW480、HT-29、HCT116)和正常人肠黏膜上皮细胞HIEC中lncRNA HSALNT0000015的表达水平;将HCT116细胞分为空白组、对照组及实验组,分别不感染慢病毒、只感染空载质粒及感染sh-HSALNT0000015-慢病毒,采用MTT法检测细胞增殖能力,Transwell实验检测细胞迁移和侵袭能力,免疫印迹法检测转录因子Slug、上皮钙黏素(E-Cadherin)及波形蛋白(Vimentin)的表达。结果结直肠癌组织lncRNA HSALNT0000015表达水平高于癌旁组织(P<0.05)。结直肠癌细胞中lncRNA HSALNT0000015的表达水平均高于正常人肠黏膜上皮细胞(P<0.05)。细胞培养12、24、48及72 h后,实验组光密度值(490 nm)低于空白组和对照组(均P<0.05)。敲减lncRNA HSALNT0000015后,HCT116迁移及侵袭能力下降(均P<0.05),E-cadherin表达水平上升(P<0.05),转录因子Slug和Vimentin表达水平下降(均P<0.05)。结论lncRNA HSALNT0000015可能通过上皮间质转化过程促进结直肠癌细胞迁移及侵袭。

Objective To investigate the effect of lncRNA HSALNT0000015 on the biological behavior of colorectal cancer cells.Methods Quantitative real-time PCR(qRT-PCR)was used to detect the expression level of lncRNA HSALNT0000015 in 80 samples of colorectal cancer tissue and 15 adjacent tissues.And in colorectal cancer cell lines Lovo,SW480,HT-29,HCT116,and normal human mammary epithelium cell line HIEC.The HCT116 cells were divided into negative control(NC)group,negative shRNA-Lenti(vector group)or Lenti-sh-HSALNT0000015(sh-HSALNT0000015 group).The cell proliferation ability was determined by MTT assay experiment;migration and invasion ability were measured by transwell assay;the transcription factor Slug,epithelial marker E-cadherin and interstitial marker vimentin were dectected by western blotting.Results The expression of HSALNT0000015 in the colorectal cancer tissues was significantly higher than that in the adjacent tissues(P<0.05).The relative expression levels of lncRNA HSALNT0000015 in colorectal cancer cell lines were significantly higher than that in normal human mammary epithelium cell line(P<0.05).MTT assay experiment demonstrated that the proliferation ability of sh-HSALNT0000015 group was lower than NC group and sh-vector group 12,24,48 and 72 h after cell culture(P<0.05).After HSALNT0000015 was knocked down,transwell assay showed that the migration and invasion ability of sh-HSALNT0000015 group was lower than NC group and sh-vector group(P<0.05)the epithelial marker E-cadherin in HCT116 was increased significantly(P<0.05),but the transcription factor Slug and interstitial marker vimentin decreased significantly(P<0.05).Conclusion lncRNA HSALNT0000015 may promote invasion and migration of colorectal cancer cells through epithelial-mesenchymal transition(EMT)process.