下调LSINCT5对人脑胶质瘤细胞系U251增殖、迁移和放疗敏感性的影响

摘要

目的 观察下调LSINCT5对人脑胶质瘤细胞系U251增殖、迁移能力和放疗敏感性的影响.方法 采用慢病毒转染法构建稳定干扰LSINCT5的细胞株(U251 sh LSINCT5细胞),得到干扰效率高的sh LSINCT51#组、sh LSINCT53#组和转染GV248的对照组细胞进行后续实验.采用噻唑蓝比色法(MTS)观察细胞培养0、24、48、72 h时增殖能力变化,划痕修复实验观察细胞迁移能力的变化,Western blotting法检测细胞中细胞黏附分子(E-cad-herin)、紧密连接蛋白-1(Zo-1)、细胞骨架蛋白(Vimentin)表达,MTS法观察X射线仪下分别照射0、2、4、8 Gy后细胞增殖能力.结果 sh LSINCT51#组、sh LSINCT53#组培养48、72 h时细胞增殖能力均较对照组低(P均<0.05).sh LSINCT51#组、sh LSINCT53#组细胞的修复距离均较对照组短,P均<0.05.与对照组相比,sh LSINCT51#组、sh LSINCT53#组E-cadherin、Zo-1表达高,Vimentin表达低(P均<0.05).sh LSINCT51#组、sh LSINCT53#组在2、4 Gy剂量照射时细胞增殖能力均较对照组低(P均<0.05).结论 下调人胶质瘤细胞系U251中LSINCT5的表达,能抑制细胞增殖、转移,并增强细胞放疗敏感性.%Objective To investigate the effect of down-regulation of LSINCT5 on proliferation,migration ability,and radiosensitivity of human glioma cell line U251. Methods Lentiviral transfection was used to construct stable LSINCT5 in-terference cells (U251 shLSINCT5 cells),and the cells in the sh LSINCT51# group and sh LSINCT53# group with high efficiency of interference and the cells in the control group (transfected with GV248) were used for the subsequent experi-ments. MTS method was used to detect the changes of cell proliferation abilities at 0,24,48,and 72 h. Scratch healing assay was used to detect the changes of cell migration abilities. Western blotting was used to detect the expression of E-cad-herin,Zo-1 and Vimentin. MTS assay was used to detect the changes of cell proliferation abilities at 0,2,4,8 Gy under X-ray. Results The proliferation abilities of the sh LSINCT51# group and sh LSINCT53# group were significantly lower than that of the control group at 48 and 72 h (all P < 0. 05). The repair distance of the sh LSINCT51# and sh LSINCT53#groups were significantly lower than that of the control group (P < 0. 05). Compared with the control group,the expression levels of E-cadherin and Zo-1 in the sh LSINCT51# group and sh LSINCT53# group increased,and the expression of Vim-entin decreased (all P < 0. 05). The proliferation abilities of sh LSINCT51# group and sh LSINCT53# group were lower than that of the control group at 2 and 4 Gy doses (both P < 0. 05). Conclusion Down-regulation of LSINCT5 expression in human glioma cell line U251 can inhibit cell proliferation and metastasis,and increase radiosensitivity of cells.