小干扰RNA沉默垂体瘤转化基因1对人胶质瘤U251细胞增殖侵袭的影响

Effect of silencing pituitary tumor transforming gene 1 by siRNA on proliferation and invasion of human glioma U251 cells

目的 探讨小干扰RNA沉默垂体瘤转化基因l (PTTG1)对人胶质瘤U251细胞增殖侵袭的影响及可能机制。方法 设计靶向沉默PTTG1的siRNA,将U251细胞随机分为PTTG1基因沉默组、阴性对照组和空白组,PTTG1基因沉默组U251细胞转染PTTG1-siRNA,阴性对照组细胞转染无关序列,空白组细胞不转染任何序列。转染后继续培养48 h,采用real-time PCR与Western blot方法鉴定转染结果,MTT方法和transwell实验分别检测细胞的增殖能力与侵袭能力,Western blot方法检测U251细胞中p-Akt、基质金属蛋白酶2/9(MMP-2/9)的表达。结果 PTTG1基因沉默组U251细胞中PTTG1 mRNA和蛋白表达显著低于阴性对照组和空白组(P<0.01),结果提示转染成功。与空白对照组及阴性对照组比较,PTTG1基因沉默组U251细胞活力显著降低,侵袭能力显著降低,p-Akt、MMP-2/9的表达显著降低(P<0.01)。结论 沉默PTTG1基因可以抑制U251细胞的增殖和侵袭,PTTG1有望成为胶质瘤靶向治疗的新靶点。

Objective To investigate the effect of small interfering RNA silencing pituitary tumor transforming gene 1(PTTG1) on the proliferation and invasion of human glioma U251 cells and its possible mechanism. Methods The siRNA targeting PTTG1 was designed, and U251 cells were randomly divided into PTTG1 gene silencing group, negative control group and blank group. U251 cells in PTTG1 gene silencing group were transfected with PTTG1 siRNA, negative control group were transfected with unrelated sequences, blank group were not transfected with any sequence. After 48 hours of culture, the transfection results were identified by real-time PCR and Western blot. The proliferation and invasion of U251cells were detected by MTT assay and Transwell assay, and the expression of p-Akt and matrix metalloproteinase-2/9(MMP-2/9) in U251 cells were detected by western blot. Results The expression of PTTG1 mRNA and protein in U251cells in PTTG1 gene silencing group was significantly lower than that in negative control group and blank group(P<0.01),which indicated that the transfection was successful. Compared with the blank control group and negative control group, the viability and invasive ability of U251 cells in PTTG1 gene silencing group were significantly decreased, and the expressions of p-Akt and MMP-2/9 were significantly decreased(P<0.01). Conclusion Silencing PTTG1 gene can inhibit the proliferation and invasion of U251 cells, and PTTG1 is expected to become a new target for targeted therapy of glioma.