长链非编码RNA PTENP1在非小细胞肺癌中的表达及对增殖和迁移的影响

Expression of long-chain non-coding RNA PTENP1 in non-small cell lung cancer and its effect on proliferation and migration

目的研究非小细胞肺癌(NSCLC)细胞系和正常肺上皮细胞系中长链非编码RNA PTENP1(LncRNA PTENP1)的表达,探讨过表达LncRNA PTENP1对NSCLC细胞增殖和迁移的影响并阐述其机制。方法采用实时荧光定量PCR(qRT-PCR)技术检测NSCLC细胞系A549、H1299、H1650、HCC827和正常肺上皮细胞系BEAS-2B中LncRNA PTENP1的表达。NSCLC细胞系H1299分别转染LncRNA PTENP1过表达质粒(PTENP1组)、阴性对照质粒(Vector组)及空白质粒(Blank组),CCK-8法和细胞划痕实验分别测定细胞增殖和迁移能力,Western blot检测PTEN和SOCS6蛋白的表达。结果NSCLC细胞系A549、H1299、H1650、HCC827中LncRNA PTENP1的相对表达水平明显低于人正常肺上皮细胞系BEAS-2B(P<0.05)。在细胞铺板后0、24、48 h,PTENP1组与Vector组450 nm处吸光度值(A450值)无明显差异(P>0.05);72、96 h,PTENP1组A450值明显低于Vector组(P<0.05)。PTENP1组划痕愈合率为(29.5±2.6)%,Vector组为(53.4±4.8)%,Blank组为(52.7±5.3)%,PTENP1组划痕愈合率明显低于Vector组(P<0.01),Vector组与Blank组无明显差异(P>0.05)。PTENP1组PTEN、SOCS6蛋白相对表达水平高于Vector组(P<0.01),Blank组与Vector组无明显差异(P>0.05)。结论LncRNA PTENP1在NSCLC细胞系中低表达,过表达LncRNA PTENP1可抑制NSCLC增殖和迁移,其机制可能与PTEN、SOCS6蛋白上调有关。

Objective To study the expression of long-chain non-coding RNA PTENP1(LncRNA PTENP1)in non-small cell lung cancer(NSCLC)cell lines and normal lung epithelial cell lines,and to explore the effect of over-expression of LncRNA PTENP1 on the proliferation and migration of NSCLC cells and elucidate its mechanism.Methods The expression of LncRNA PTENP1 in NSCLC cell lines A549,H1299,H1650,HCC827 and normal lung epithelial cell line BEAS-2B was detected by real-time fluorescence quantitative PCR(qPCR).LncRNA PTENP1 overexpression plasmid(the PTENP1 group),negative control plasmid(the vector group)and blank plasmid(the blank group)were transfected into NSCLC cell line H1299.Cell proliferation and migration were measured by CCK-8 method and cell scratch test.Western blot was used to analyze the expression of PTEN and SOCS6 protein.Results The relative expression level of LncRNA PTENP1 in NSCLC cell lines A549,H1299,H1650 and HCC827 was significantly lower than that in human normal lung epithelial cell lines BEAS-2B(P<0.05).There was no significant difference in value of absorbance at 450 nm(A450)between the PTENP1 group and the vector group at 0,24 and 48 h after cell placement(P>0.05);while,the of A450 in the PTENP1 group was significantly lower than that in the vector group at 72 and 96 h after cell placement(P<0.05).The healing rate of scratches in the PTENP1 group was(29.5±2.6)%,(53.4±4.8)%in the vector group,and(52.7±5.3)%in the blank group.The healing rate of scratches in the PTENP1 group was significantly lower than that in the vector group(P<0.01),while no statistically significant difference was found between the vector group and the blank group(P>0.05).The relative expression levels of PTEN and SOCS6 proteins in the PTENP1 group were higher than those in the vector group(P<0.05),while no statisticallysignificant difference was found between the blank group and the vector group(P>0.05).Conclusion LncRNA PTENP1 is under-expressed in NSCLC cell lines.Overexpression of LncRNA PTENP1 can inhibit the proliferation and migration of NSCLC cells,which may be related to the up-regulation of PTEN and SOCS6 proteins.