人参皂苷Rh2调节转化生长因子β活化的长链非编码RNA对胶质瘤细胞侵袭和迁移的影响

Effects of ginsenoside Rh2 on proliferation,migration and invasion of glioma cells through regulating long non-coding RNA-activated by transforming growth factor-beta

目的探讨人参皂苷Rh2(G-Rh2)调控转化生长因子β活化的长链非编码RNA(LncRNA-ATB)对胶质瘤细胞增殖、迁移和侵袭的影响。方法采用实时荧光定量PCR(qPCR)检测人星形胶质细胞NHA和胶质瘤细胞(SHG-44、A172、Ln229、U87、U251)的LncRNA-ATB水平。培养U251细胞,将细胞分为si-NC组(阴性对照)和si-ATB组(干扰LncRNA-ATB),采用CCK-8、Transwell小室实验和划痕实验分别评价细胞增殖、迁移和侵袭能力。不同浓度G-Rh2处理U251细胞以计算G-Rh2的半数抑制浓度(IC_(50))值。将细胞分为对照组、G-Rh2组(20μg/ml G-Rh2)和G-Rh2+ATB组(20μg/ml G-Rh2+过表达LncRNA-ATB),检测细胞增殖、迁移和侵袭情况,qPCR和Western blotting检测基质金属蛋白酶(MMP)-9和Snail的水平。结果LncRNA-ATB在胶质瘤细胞的水平低于NHA细胞(P<0.05)。与si-NC组相比,si-ATB组U251细胞增殖、迁移和侵袭能力均受抑制(P<0.05)。G-Rh2的IC_(50)值为20.992μg/ml,并能够抑制U251细胞中LncRNA-ATB的表达(P<0.05)。G-Rh2组U251细胞增殖活力、迁移和侵袭能力低于对照组;G-Rh2+ATB组U251细胞增殖活力、迁移和侵袭能力明显高于G-Rh2组。G-Rh2组MMP-9和Snail的水平低于对照组(P<0.05),G-Rh2+ATB组MMP-9和Snai水平高于G-Rh2组(P<0.05)。结论干扰LncRNA-ATB表达抑制U251细胞的增殖、迁移和侵袭过程,G-Rh2可能下调LncRNA-ATB表达来抑制U251细胞的增殖、迁移和侵袭,发挥抗肿瘤功效。

Objective To explore the effects of ginsenoside Rh2(G-Rh2)on proliferation,migration and invasion of glioma cells through regulating long non-coding RNA-activated by transforming growth factor-beta(LncRNA-ATB).Methods Real-time quantitative polymerase chain reaction(qPCR)was performed to detect the level of LncRNA-ATB in human astrocytes NHA cell and glioma cells(SHG-44,A172,Ln229,U87 and U251).U251 cells were cultured and divided into si-NC group(negative control)and si-ATB group(downregulation of LncRNA-ATB).The proliferation,migration and invasion abilities of U251 cells were assessed by CCK-8,wound-healing and Transwell assays.U251 cells were treated with G-Rh2,and half-maximal inhibitory concentration(IC_(50))value of G-Rh2 was calculated.U251 cells were divided into Control group,G-Rh2 group(20μg/ml G-Rh2)and G-Rh2+ATB group(20μg/ml G-Rh2 plus overexpression of LncRNA-ATB).The cell proliferation,migration and invasion were measured.QPCR and Western blotting were used to examine the levels of matrix metalloproteinase(MMP)-9 and Snail.Results The levels of LncRNA-ATB in glioma cells were higher than NHA cells(P<0.05).The cell proliferation,migration and invasion of si-ATB group were inhibited compared with si-NC group.The IC_(50) value of G-Rh2 in U251 cells was 20.992μg/ml.G-Rh2 inhibited LncRNA-ATB expression in U251 cells(P<0.05).The cell proliferation,migration and invasion of G-Rh2 group were decreased compared with Control group(P<0.05).While the cell proliferation,migration and invasion of G-Rh2+ATB group were increased compared with G-Rh2 group.Moreover,the levels of MMP-9 and Snail were lower in G-Rh2 group than those in Control group(P<0.05).Levels of MMP-9 and Snail were higher in G-Rh2+ATB group than those in G-Rh2 group(P<0.05).Conclusion Silence of LncRNA-ATB inhibited proliferation,migration and invasion of glioma cell U251.G-Rh2 might suppresses the proliferation,migratory and invasive abilities of U251 cells possibly by downregulating LncRNA-ATB,exerting anti-tumor effects in glioma.