MiR497HG靶向miR-588调控肺癌细胞增殖、迁移侵袭和凋亡的机制研究

目的研究MiR497HG对肺癌细胞增殖、迁移侵袭及凋亡的影响,并探讨其机制。方法运用RT-PCR法检测永生化正常肺上皮细胞BEAS-2 B、肺癌细胞NCI-H1975、NCI-H460、A549中MiR497HG、miR-588的表达;将blank组(不做任何处理)、si-NC组(转染si-NC)、si-MiR497HG组(转染si-MiR497HG)、anti-miR-NC组(转染anti-miR-NC)、anti-miR-588组(转染anti-miR-588)、miR-NC组(转染miR-NC)、miR-588组(转染miR-588 mimics)、si-MiR497HG+anti-miR-NC组(共转染si-MiR497HG和anti-miR-NC)、si-MiR497HG+anti-miR-588组(共转染si-MiR497HG和anti-miR-588),均用脂质体法转染至NCI-H1975细胞;MTT法检测细胞增殖;Transwell小室检测细胞的迁移侵袭;流式细胞术检测细胞凋亡;双荧光素酶报告基因检测实验检测细胞中MiR497HG与miR-588的结合力。结果与永生化正常肺上皮细胞BEAS-2 B相比,肺癌细胞NCI-H1975、NCI-H460、A549中MiR497HG表达显著升高,miR-588表达显著降低(P<0.05);抑制MiR497HG或过表达miR-588均可抑制NCI-H1975细胞的增殖、迁移侵袭,促进凋亡;miR-588可抑制野生型MiR497HG细胞的荧光活性,且可负向调控MiR497HG的表达;过表达MiR497HG可逆转miR-588对肺癌细胞的增殖、迁移侵袭抑制和凋亡促进作用。结论长链非编码RNA MiR497HG可促进肺癌细胞增殖、迁移侵袭,抑制凋亡,其机制可能与靶向抑制miR-588有关,将可为肺癌的治疗提供新靶点。

Mechanism of MiR497HG regulating cell proliferation,migration, invasion and apoptosis of lung cancer cells by targeting miR-588

Objective To study the effect of MiR497HG on proliferation,migration,invasion and apoptosis of lung cancer cells,and to explore its mechanism.Methods RT-PCR was used to detect the expression of MiR497HG and miR-588 in immortalized normal lung epithelial cells BEAS-2 B,lung cancer cells NCI-H1975,NCI-H460 and A549.They were divided into the blank group(without any treatment),the si-NC group(transfected si-NC),the si-MiR497HG group(transfected si-MiR497HG),the anti-miR-NC group(transfected anti-miR-NC),the anti-miR-588 group(transfected anti-miR-588),the miR-NC group(transfected miR-NC),the miR-588 group(transfected miR-588 mimics),the si-MiR497HG+anti-miR-NC group(co-transfected si-MiR497HG and anti-miR-NC),and the si-MiR497HG+anti-miR-588 group(co-transfected si-MiR497HG and anti-miR-588).All were transfected into NCI-H1975 cells by liposome method.Cell proliferation was detected by MTT assay.Transwell was used to detect cell migration and invasion.Flow cytometry was used to detect apoptosis.Dual luciferase reporter assay was used to detect the binding of MiR497HG to miR-588 in cells.Results Compared with the immortalized normal lung epithelial cells BEAS-2B,the expression of MiR497HG was significantly increased in lung cancer cells NCI-H1975,NCI-H460 and A549,and the expression of miR-588 was significantly decreased(P<0.05).MiR497HG or over-expression was inhibited.miR-588 could inhibit the proliferation,migration and invasion of NCI-H1975 cells and promote apoptosis.miR-588 could inhibit the fluorescence activity of wild-type MiR497HG cells and negatively regulate the expression of MiR497HG.The over-expression of MiR497HG could reverse miR-588 proliferation,migration and invasion inhibition and apoptosis promotion of lung cancer cells.Conclusion The long-chain non-coding RNA MiR497HG can promote the proliferation,migration and inhibition of lung cancer cells,and its mechanism may be related to the targeted inhibition of miR-588,which will open a new target for the treatment of lung cancer.