羽扇豆醇调控miR-100-5p对非小细胞肺癌NCI-H1299细胞生物学行为的影响

Effects of lupeol on the biological behavior of non-small cell lung cancer NCI-H1299 cells by regulating miR-100-5p

目的 探讨羽扇豆醇调控miR-100-5p对非小细胞肺癌(NSCLC)NCI-H1299细胞增殖、迁移和侵袭的影响。方法 将体外培养的NCI-H1299细胞分为正常对照(NC)组(未处理)、低剂量组(12.5μmol/L羽扇豆醇)、中剂量组(25μmol/L羽扇豆醇)、高剂量组(50μmol/L羽扇豆醇)、miR-NC组(转染miR-NC)、miR-100-5p组(转染miR-100-5p)、anti-miR-NC+高剂量组(转染anti-miR-NC后用50μmol/L羽扇豆醇处理)、anti-miR-100-5p+高剂量组(转染anti-miR-100-5p后用50μmol/L羽扇豆醇处理)。CCK-8和克隆形成实验检测NCI-H1299细胞增殖;Transwell法检测NCI-H1299细胞迁移和侵袭。荧光定量PCR(q PCR)检测miR-100-5p表达;Western blot检测细胞周期蛋白D1(CyclinD1)、基质金属蛋白酶(MMP)2和MMP9蛋白的表达。结果 与NC组比较,低、中、高剂量组NCI-H1299细胞活力、细胞克隆数、迁移数、侵袭数、CyclinD1、MMP2和MMP9蛋白表达降低(P<0.05),miR-100-5p表达逐渐升高(P<0.05)。与miR-NC组比较,miR-100-5p组miR-100-5p表达升高,NCI-H1299细胞活力、细胞克隆数、迁移数、侵袭数、CyclinD1、MMP2和MMP9蛋白表达降低(P<0.05);与anti-miR-NC+高剂量组比较,anti-miR-100-5p+高剂量组miR-100-5p表达下降,NCI-H1299细胞活力、细胞克隆数、迁移数、侵袭数、CyclinD1、MMP2和MMP9蛋白表达升高(P<0.05)。结论 羽扇豆醇通过上调miR-100-5p表达抑制NSCLC细胞NCI-H1299增殖、迁移和侵袭。

Objective To investigate the effect of lupeol on the proliferation,migration and invasion of non-small cell lung cancer(NSCLC) NCI-H1299 cells by regulating miR-100-5p.Methods NCI-H1299 cells cultured in vitro were divided into the normal control(NC) group(untreated),the low-dose group(12.5 μmol/L lupeol),the medium-dose group(25 μmol/L lupeol),the high-dose group(50 μmol/L lupeol),the miR-NC group(transfected with miR-NC),the miR-100-5p group(transfected with miR-100-5p),the anti-miR-NC+high-dose group(the cells were treated with 50 μmol/L lupeol after transfected with anti-miR-NC),and the anti-miR-100-5p+high-dose group(the cells were treated with 50 μmol/L lupeol after transfected with anti-mi R-100-5 p).CCK-8 and colony formation assays were performed to evaluate the proliferation of NCI-H1299 cells.Transwell method was applied to detect the migration and invasion of NCI-H1299 cells.RT-qPCR was used to detect the expression of miR-100-5p.Western blot assay was used to detect the expression of CyclinD1,matrix metalloproteinase 2(MMP2) and matrix metalloproteinase 9(MMP9) proteins.Results Compared with the NC group,NCI-H1299 cell viability,colony formation number,migration number,invasion number,the expression levels of CyclinD1,MMP2 and MMP9 proteins were decreased in the low,medium and high dose groups(P<0.05),and the expression of miR-100-5p was significantly increased(P <0.05).Compared with the miR-NC group,the expression of miR-100-5p was increased,the NCI-H1299 cell viability,colony formation number,migration number,invasion number,the expression of CyclinDl,MMP2 and MMP9 proteins decreased in the miR-100-5p group(P <0.05).Compared with the antimiR-NC+high-dose group,the expression of miR-100-5p was decreased,NCI-H1299 cell viability,colony formation number,migration number,invasion number,the expression of CyclinDl,MMP2 and MMP9 proteins increased in the antimiR-100-5p+high-dose group(P <0.05).Conclusion Lupeol inhibits the proliferation,migration and invasion of NSCLC cells NCI-H1299 by up-regulating miR-100-5p.