长链非编码RNA LINC00324对非小细胞肺癌细胞生物学功能的影响

Effect of long non-coding RNA LINC00324 on biological functions of non-small cell lung cancer cells

目的观测长链非编码RNA LINC00324在非小细胞肺癌中的表达情况,探讨其对非小细胞肺癌细胞系生物学功能的影响。方法通过qRT-PCR检测LINC00324在65对配对肺癌组织及癌旁正常组织中的表达,检测不同非小细胞肺癌细胞系(4株非小细胞肺癌细胞系:A549、SPCA1、H460和H1299; 1株支气管上皮细胞系:HBE)中LINC00324基因的表达;构建LINC00324基因的过表达载体,构建LINC00324过表达细胞系;采用CCK-8、Transwell、流式细胞术检测过表达LINC00324基因后对非小细胞肺癌细胞增殖、侵袭、迁移及凋亡的影响。结果 LINC00324在肺癌组织中较癌旁正常组织低表达(P <0. 05);以HBE作为对照,LINC00324在A549和H460细胞系中相对低表达(P <0. 05),在H1299和SPCA1细胞系中相对高表达(P <0. 05);成功构建LINC00324过表达A549和H460细胞系,LINC00324基因在A549和H460过表达细胞系中表达均升高(P <0. 05); CCK-8实验显示,过表达LINC00324基因能够抑制H460和A549肺癌细胞的增殖(P <0. 05); Transwell实验显示,过表达LINC00324基因较空质粒对照组能够显著降低H460和A549肺癌细胞的侵袭和迁移(P <0. 05);流式细胞术检测结果显示,过表达LINC00324基因较空质粒对照组能够显著增加H460和A549肺癌细胞的凋亡(P <0. 05)。结论长链非编码RNA LINC00324可能通过调节肺癌细胞的增殖、迁移、侵袭和凋亡,在肺癌的发生、发展中起着重要的抑癌基因作用。

Objective To investigate the expression of long non-coding RNA LINC00324 in non-small cell lung cancer( NSCLC) and explore its effect on the biological functions of the tumor cells.Methods q RTPCR was used to detect the expression of LINC00324 in 65 paired lung cancer tissues and adjacent normal tissues.The expression of LINC00324 was also detected in 4 different non-small cell lung cancer cell lines( A549,SPCA1,H460 and H1299) and human bronchial epithelial cell line HBE by qRT-PCR.The overexpression vector of LINC00324 gene was designed and constructed,and the LINC00324 overexpression cell lines were constructed through transfection.The effects of overexpression of LINC00324 on the proliferation,invasion, migration and apoptosis of NSCLC cells were measured by cell counting kit-8( CCK-8) assay,Transwell assay,and flow cytometry respectively.Results LINC00324 was down-regulated in the lung cancer tissues than adjacent normal tissues( P< 0.05).The expression of LINC00324 was observed in 4 NSCLC lines and HBE cells.The expression of LINC00324 was lowest in A549 and H460 cells( P< 0.05),and highest in H1299 and SPCA1 cells( P< 0.05).LINC00324 overexpression was successfully constructed in A549 and H460 cells.The expression of LINC00324 gene was increased in both A549 and H460 overexpressed cell lines( P< 0.05).The overexpression of LINC00324 gene inhibited the proliferation( P< 0.05),reduced the invasion and migration( P< 0.05),and promoted apoptosis( P< 0.05)in the H460 and A549 cells,when compared with the cells transfected with blank plasmids.Conclusion Long non-coding RNA LINC00324 may play an important anti-oncogene role in the development and progression of lung cancer by regulating the proliferation,migration,invasion,and apoptosis of lung cancer cells.