敲低lncRNA DILC对胶质瘤细胞体外生物学特性的影响

Effects of knocking down of lncRNA DILC on the biological characteristics of glioma cells in vitro

目的探讨敲低长链非编码RNA(lncRNA)对胶质瘤细胞体外生物学特性的影响。方法胶质瘤细胞系LN382、U87MG分别分组为NC组和敲低组,其中NC组以negative siRNA转染,敲低组以DILC siRNA转染。转染前和转染后48 h,PCR检测IL-6、JAK2、STAT3表达水平,ELISA法检测离心细胞上清液中IL-6水平,Western Blot法检测p-JAK2/JAK2和p-STAT3/STAT3蛋白质表达水平。细胞转染后48 h,Transwell小室侵袭实验观察细胞的侵袭能力,划痕实验观察细胞的转移能力状况。结果细胞转染后48 h,与LN382、U87MG胶质瘤细胞的NC组比较,其相应敲低组的穿膜细胞数增加,细胞迁移距离亦增加(P<0.05)。与LN382、U87MG胶质瘤细胞的NC组比较,其相应敲低组的IL-6、JAK2、STAT3的mRNA相对表达量升高,细胞上清液中IL-6水平升高,p-JAK2/JAK2和p-STAT3/STAT3蛋白质表达水平亦相应升高(P<0.05)。结论敲低lncRNA DILC可促进胶质瘤细胞的侵袭转移,而激活IL-6/JAK2/STAT3信号通路可能为其作用机制。

Objective To investigate the effect of knocking down long-chain non coding RNA(lncRNA)on the biological characteristics of glioma cells in vitro. Methods Glioma cell lines ln382 and U87 MG were both divided into two groups:NC group and knocking down group. The NC group was transfected with negative siRNA,while the knocking down group was transfected with dilc siRNA. Before and48 hours after transfection,the expression levels of IL-6,JAK2 and STAT3 were measured by RT-qPCR. The IL-6 level in the cell culture supernatant was detected by ELISA. The protein levels of p-JAK2/JAK2 and pSTAT3/STAT3 were detected by Western blot. 48 hours after transfection,the Transwell cell invasion experiment was used to observe the cell invasion ability,and the scratch experiment was used to observe the cell migration ability status. Results At 48 h after cell transfection,compared with the NC group of LN382 and U87 MG glioma cells,the number of transmembrane cells in the corresponding knocking down group increased,and the cell migration distance also increased(P<0.05). Compared with the NC group of LN382 and U87 MG glioma cells,the relative expression levels of IL-6,JAK2,and STAT3 mRNA in the corresponding knocking down group increased,and the IL-6 level in the supernatant increased. p-JAK2/JAK2 and p-STAT3/STAT3 protein expression levels also increased accordingly(P<0.05). Conclusion Knocking down lncRNA DILC can promote the invasion and metastasis of glioma cells. The mechanism of action may relate to the activation of IL-6/JAK2/STAT3 signaling pathway.