外源性p16基因稳定转染对胶质瘤细胞周期和增殖的影响

目的：研究外源性ｐ１６基因对胶质瘤细胞系ＳＨＧ－４４的抑制作用，探索胶质瘤基因治疗的新途径。方法：利用携带４９０ｂｐ的人ｐ１６全长ｃＤＮＡ真核表达载体，采用脂质体介导法将其导入人恶性胶质瘤细胞系ＳＨＧ－４４中，用Ｇ４１８筛选阳性克隆，以原位杂交，免疫组化方法检测ｐ１６基因在细胞转染前后的表达情况。，应用流式细胞仪，电子显微镜，生长曲线测定等方法研究ｐ１６基因对胶质瘤细胞周期，形态，生长等的影响.

Effects of exogenous p16/CDKN2 gene on the cell cycle and proliferation of SHG-44 glioma cell line

AIM: To investigate the suppressive effects of exogenous p16/CDKN2 gene on SHG44 glioma cell line and to probe into new methods of gene therapy for glioma. METHODS: A eukaryotic expression vector containing 490 bp human fulllength p16/CDKN2 cDNA was transfected into human SHG44 glioma cells using lipofectamine. Expression of exogenous p16/CDKN2 gene was detected by in situ hybridization and immunohistochemistry. Flow cytometry, electron microscope and other methods were adopted to measure the effects of exogenous p16/CDKN2 gene on cell cycle progression, morphology and cell growth features of the transfected glioma cells, as well as on tumorigenicity in nude mice. RESULTS: In situ hybridization and immunohistochemistry revealed that p16/CDKN2 mRNA and protein were expressed in the transfected glioma cells. The progression of cell cycle was arrested from G1 to S phase. The growth of cells transfected with p16/CDKN2 cDNA in nude mice was dramatically inhibited compared to the parent SHG44 cells. CONCLUSION: Glioma cells could be arrested at G1/S phase and the growth of cells could be significantly suppressed by exogenous p16/CDKN2 gene.