lncRNA PCAT19吸附miR-142-5p调控ING3基因表达抑制鼻咽癌细胞的增殖和侵袭

lncRNA PCAT19 inhibits the proliferation and invasion of nasopharyngeal carcinoma cells by adsorbing miR-142-5p and regulating the expression of ING3 gene

目的:检测长链非编码RNA(long non-coding RNA,lncRNA)PCAT19在鼻咽癌组织和细胞株中的表达量,探讨其抑制鼻咽癌细胞增殖和侵袭的分子作用机制。方法:实时荧光定量PCR(qRT-PCR)法检测鼻咽癌组织和5种鼻咽癌细胞株中PCAT19的相对表达。在表达最低的鼻咽癌细胞株转染空载质粒(对照组)或高表达PCAT19质粒(实验组)。qRT-PCR检测转染效率。MTS法和Transwell侵袭实验分别检测高表达PCAT19对鼻咽癌细胞增殖能力和侵袭能力的影响。生物信息学预测PCAT19的靶基因。qRT-PCR和Western blot检测靶基因在mRNA和蛋白水平的表达。结果:与慢性鼻咽炎组织比较,PCAT19在鼻咽癌组织的表达降低(P<0.01)。与永生化鼻咽上皮细胞比较,5种鼻咽癌细胞株PCAT19的表达均明显降低(P<0.05),SUNE-1细胞中的表达最低(P<0.01)。转染高表达PCAT19质粒可明显促进PCAT19的表达(P<0.01)。高表达PCAT19可抑制鼻咽癌SUNE-1细胞的增殖能力(P<0.01)和侵袭能力(P<0.01)。PCAT19的靶基因是miR-142-5p,miR-142-5p的靶基因生长抑制因子3(inhibitor of growth gene 3,ING3)。高表达PCAT19后,miR-142-5p表达明显降低(P<0.01),而ING3在mRNA和蛋白水平上的表达均明显增加(P<0.01)。结论:鼻咽癌组织和细胞株中PCAT19的表达下调。上调PCAT19可抑制鼻咽癌SUNE-1细胞的增殖和侵袭能力,其作用机制可能是PCAT19通过吸附miR-142-5p促进ING3基因的表达。

AIM:To investigate the expression of long non-coding RNA(lncRNA)PCAT19 in nasopharyngeal carcinoma tissues and cell lines,and to explore its molecular mechanism of inhibiting proliferation and invasion of nasopharyngeal carcinoma cells.METHODS:Real-time fluorescent quantitative PCR(qRT-PCR)was used to detect the relative expression of PCAT19 in nasopharyngeal carcinoma tissues and five nasopharyngeal carcinoma cell lines.The lowest expressing nasopharyngeal carcinoma cell line was transfected with empty plasmid(control group)or high expression PCAT19 plasmid(experimental group).qRT-PCR was used to detect transfection efficiency.MTS method and Transwell invasion test were used to detect the effect of overexpressing PCAT19 on the proliferation and invasion ability of nasopharyngeal carcinoma cells.Bioinformatics predicts target genes for PCAT19.qRT-PCR and Western blot were used to detect target gene expression at mRNA and protein levels.RESULTS:Compared with adjacent tissues,the expression of PCAT19 in nasopharyngeal carcinoma tissues was decreased(P<0.01).Compared with immortalized nasopharyngeal epithelial cells,the expression of PCAT19 in five nasopharyngeal carcinoma cell lines was significantly reduced(P<0.05),and the expression was the lowest in SUNE-1 cells(P<0.01).Transfection of high-expressing PCAT19 plasmid could significantly promote the expression of PCAT19(P<0.01).High expression of PCAT19 could inhibit the proliferation ability(P<0.01)and invasive ability(P<0.01)of nasopharyngeal carcinoma SUNE-1 cells.The target gene of PCAT19 was miR-142-5 p,the target gene of miR-142-5 p was inhibitor of growth gene 3(ING3).After high expression of PCAT19,miR-142-5 p expression was significantly reduced(P<0.01),and the expression of ING3 at both mRNA and protein levels was significantly increased(P<0.01).CONCLUSION:PCAT19 expression is down-regulated in nasopharyngeal carcinoma tissues and cell lines.Up-regulating PCAT19 can inhibit the proliferation and invasion of nasopharyngeal carcinoma SUNE-1 cells.The mechanism may be that PCAT19 promotes the expression of ING3 gene by adsorbing miR-142-5 p.