lncRNA GPC3-AS1在非小细胞肺癌中的表达及对癌细胞生物学行为的影响

Expression of lncRNA GPC3-AS1 in non-small cell lung cancer and its effect on the biological behavior of the cancer cells

目的探讨lncRNA GPC3-AS1在非小细胞肺癌组织中的表达及对A549细胞增殖、迁移和侵袭的影响。方法qPCR检测非小细胞肺癌组织和细胞中lncRNA GPC3-AS1表达;将lncRNA GPC3-AS1 siRNA及其对照siRNA转入A549细胞中,qPCR检测lncRNA GPC3-AS1表达变化,MTT测定细胞增殖,划痕实验测定细胞迁移,Transwell测定细胞侵袭,免疫印迹检测细胞中PI3K/Akt信号通路蛋白表达。结果与非小细胞肺癌癌旁组织和人肺上皮细胞BEAS-2B相比,非小细胞肺癌组织和HCC827、H1650、A549细胞中lncRNA GPC3-AS1表达明显增加(P<0.05)。NSCLC组织中lncRNA GPC3-AS1表达与性别、年龄、病理类型及分化程度均无关,而与TNM分期、肿瘤大小、淋巴结转移均明显相关(P<0.05)。与沉默对照组相比,沉默GPC3-AS1组A549细胞中lncRNA GPC3-AS1表达显著降低(P<0.05),细胞增殖、迁移和侵袭能力亦显著降低(P<0.05)。沉默GPC3-AS1组和抑制剂组细胞中PI3K/Akt信号通路蛋白p-PI3K、p-Akt及其下游蛋白Cyclin D1和P70S6K表达较沉默对照组和对照组显著降低(P<0.05)。结论LncRNA GPC3-AS1在非小细胞肺癌中高表达,沉默LncRNA GPC3-AS1可抑制非小细胞肺癌细胞增殖、迁移和侵袭,其机制可能与抑制PI3K/Akt信号通路有关。

Objective To investigate the expression of lncRNA GPC3-AS1 in non-small cell lung cancer(NSCLC)and its effects on the proliferation,migration and invasion of A549 cells.Methods qPCR was used to detect the expression of lncRNA GPC3-AS1 in non-small cell lung cancer tissues and cells.Then,lncRNA GPC3-AS1 siRNA and its control siRNA were transferred into A549 cells.qPCR was used to detect the change in lncRNA GPC3-AS1 expression;MTT assay was used to examine the cell proliferation;scratch test was used to measure the cell migration;Transwell assay was used to measure cell invasion,and Western blotting was used to detect the expression of PI3K/Akt signaling pathway proteins in cells.Results Compared with NSCLC-adjacent tissues and human lung epithelial cells BEAS-2B,the expression of lncRNA GPC3-AS1 in NSCLC tissues and HCC827,H1650,and A549 cells was significantly increased(P<0.05).The expression of lncRNA GPC3-AS1 in NSCLC tissues was not related to gender,age,pathology and differentiation,but was significantly related to TNM staging,tumor size,and lymph node metastasis(P<0.05).Compared with the silenced control group,the expression of lncRNA GPC3-AS1 in A549 cells in the GPC3-AS1-silenced group was significantly reduced(P<0.05),and the ability of cell proliferation,migration and invasion was also significantly reduced(P<0.05).The expressions of PI3K/Akt signaling pathway proteins p-PI3K,p-Akt and its downstream proteins Cyclin D1 and P70S6K in the cells of GPC3-AS1-silenced group and inhibitor group were significantly lower than those in the silenced control group and the control group(P<0.05).Conclusion LncRNA GPC3-AS1 is highly expressed in NSCLC.Silencing LncRNA GPC3-AS1 can inhibit the proliferation,migration and invasion of NSCLC cells.The mechanism may be related to inhibition of PI3K/Akt signaling pathway.