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目的探讨PTEN基因转染人胶质瘤U251细胞后对其增殖及侵袭性的影响。方法应用脂质体转染技术,将含PTEN基因重组表达质粒转染U251细胞系,Western blot鉴定其蛋白表达;应用四甲基偶氮唑蓝比色(MTT)法观察对细胞株生长的影响;流式细胞仪检测细胞周期及凋亡;应用免疫组化检测转染PTEN的U251细胞基质金属蛋白酶(MMP-2、MMP-9)和金属蛋白酶抑制剂(TIMP-1、TIMP-2)的表达变化。结果 PTEN质粒成功转染U251细胞并有PTEN蛋白的表达。与对照组、空载体组相比,转染组细胞生长抑制率下降达52.46%;细胞周期改变,细胞从G1期到S期发生阻滞;转染组MMP-2、MMP-9表达显著下降,而TIMP-1、TIMP-2表达显著上升,两者呈负相关(r=-0.506,P<0.05)。结论 PTEN转染人胶质瘤细胞系U251后,可抑制其增殖、促进凋亡;并可能通过抑制MMPs和促进TIMPs来抑制细胞侵袭性。
Objective To investigate the effect of PTEN gene on proliferation and invasiveness of human glioma U251 cells. Methods The recombinant plasmids containing PTEN gene were transfected into U251 cell line by lipofection technique and the protein expression was determined by Western blot. The effect of PTEN gene on cell growth was observed by MTT assay The cell cycle and apoptosis were detected by flow cytometry. The expressions of MMP-2 and MMP-9 and TIMP-2 in U251 cells transfected with PTEN were detected by immunohistochemistry Change of expression Results PTEN plasmid was successfully transfected into U251 cells and expressed PTEN protein. Compared with the control group and the empty vector group, the cell growth inhibition rate decreased by 52.46% in the transfected group, the cell cycle changed, the cells from the G1 phase to the S phase block; transfected group MMP-2, MMP-9 expression was significantly decreased , While the expressions of TIMP-1 and TIMP-2 increased significantly (r = -0.506, P <0.05). Conclusion PTEN transfected human u251 glioma cell line U251 can inhibit its proliferation and promote apoptosis, and may inhibit cell invasiveness by inhibiting MMPs and promoting TIMPs.