上调lncRNA CTA-246H3.12通过调节miR-515-5p抑制鼻咽癌细胞的增殖和侵袭

Up-regulation of Long-chain Non-coding RNA CTA-246H312 Inhibits Proliferation and Invasion of Nasopharyngeal Carcinoma Cells by Modulating miR-515-5p

目的探讨长链非编码RNA(lncRNA)CTA-246H3.12在鼻咽癌组织和细胞系中的表达,研究上调CTA-246H3.12影响鼻咽癌细胞增殖和侵袭的分子机制。方法qPCR分别检测CTA-246H3.12在鼻咽癌组织和鼻咽癌细胞系中的表达。选取CTA-246H3.12表达量最低的细胞系,分别转染阴性质粒或表达CTA-246H3.12的质粒,定义为阴性对照组和CTA-246H3.12组。MTT法和Transwell侵袭实验分别检测上调CTA-246H3.12对细胞增殖和侵袭能力的影响。生物信息学方法预测CTA-246H3.12的靶基因。qPCR和Western blot法分别检测上调CTA-246H3.12对靶基因表达的影响。结果与慢性鼻咽炎组织比较,CTA-246H3.12在鼻咽癌组织表达下调(P<0.01)。与人永生化鼻咽上皮细胞比较,CTA-246H3.12在鼻咽癌细胞系表达下调(P<0.05),CTA-246H3.12在C666-1细胞中的表达量最低(P<0.01)。与阴性对照组比较,CTA-246H3.12组C666-1细胞的增殖能力显著降低(P<0.05),细胞侵袭能力显著降低(P<0.01)。CTA-246H3.12的靶基因可能是miR-515-5p,miR-515-5p的靶基因可能是尾侧型同源盒转录因子1(CDX1)。与阴性对照组比较,CTA-246H3.12组C666-1细胞中miR-515-5p表达显著降低(P<0.01),CDX1在mRNA和蛋白水平的表达显著增加。结论CTA-246H3.12在鼻咽癌组织及细胞系中表达下调,上调CTA-246H3.12可明显抑制鼻咽癌C666-1细胞的增殖和侵袭能力,其分子机制可能是通过抑制miR-515-5p的表达,间接促进CDX1基因的表达。

Objective To investigate the expression of long-chain non-coding RNA(lncRNA)CTA-246 H3.12 in nasopharyngeal carcinoma tissues and cell lines,and to investigate the molecular mechanism of up-regulation of CTA-246 H3.12 on proliferation and invasion of nasopharyngeal carcinoma cells.Methods qPCR was used to detecte the expression of CTA-246 H3.12 in nasopharyngeal carcinoma and nasopharyngeal carcinoma cell lines,respectively.The cell line with the lowest expression of CTA-246 H3.12 was selected and transfected with a negative plasmid or a plasmid expressing CTA-246 H3.12,which was defined as a negative control group and a CTA-246 H3.12 group.MTT assay and Transwell invasion assay were used to detect the effect of up-regulation of CTA-246 H3.12 on cell proliferation and invasion.The bioinformatics method predicts the target gene of CTA-246 H3.12.qPCR and Western blot were used to detect the effect of up-regulation of CTA-246 H3.12 on target gene expression.Results Compared with chronic nasopharyngitis tissues,CTA-246 H3.12 was down-regulated in nasopharyngeal carcinoma tissues(P<0.01).Compared with human immortalized nasopharyngeal epithelial cells,CTA-246 H3.12 was down-regulated in nasopharyngeal carcinoma cell lines(P<0.05),and CTA-246 H3.12 was the lowest in C666-1 cells(P<0.01).Compared with the negative control group,the proliferation ability of C666-1 cells in CTA-246 H3.12 group was significantly decreased(P<0.05),and the cell invasion ability was significantly decreased(P<0.01).The target gene of CTA-246 H3.12 may be miR-515-5 p,and the target gene of miR-515-5 p may be caudal homeobox transcription factor 1(CDX1).Compared with the negative control group,the expression of miR-515-5 p was significantly decreased in SMTA-7721 cells of CTA-246 H3.12 group(P<0.01),and the expression of CDX1 at mRNA and protein levels was significantly increased.Conclusion CTA-246 H3.12 was down-regulated in nasopharyngeal carcinoma tissues and cell lines.Up-regulation of CTA-246 H3.12 significantly inhibited the proliferation and invasion of nasopharyngeal carcinoma C666-1 cells.The molecular mechanism may be that it inhibits the expression of miR-515-5 p indirectly promotes the expression of the CDX1 gene.