腺病毒介导的p16基因对TJ899细胞生长的影响

p16/CDKN2 Gene Transfected via Adenoviral Vector Suppresses the Growth of Human Glioma Cell line TJ899

重建抑癌基因功能是肿瘤基因治疗所追求的一个基本目标。其中抑癌基因p16的缺失、失活、突变等改变在多种肿瘤中发生率较高。尤其是在人脑恶性胶质瘤中p16的缺失更为常见。本实验研究5型腺病毒载体(Ad5)携带p16基因对恶性脑胶质瘤细胞系TJ899生长状态的影响。方法:采用免疫组化(SP法)测定p16蛋白表达,MTT(methly thiazolyl tetrazolium,MTT)法测定恶性脑胶质瘤细胞系生长状态,克隆形成实验。结果:重组体腺病毒能介导p16外源基因在恶性脑胶质瘤细胞系TJ899细胞中表达,6天时肿瘤细胞生长抑制率为93%,并且能显著地抑制肿瘤细胞的克隆形成能力。结论:腺病毒介导p16基因在肿瘤细胞中表达能明显抑制肿瘤细胞的生长。

BACKGROUND & OBJECTIVE: Both the activation of proto-oncogene and the inactivation of tumor suppressor gene are the fundamental steps in the development of tumor. Particularly, the latter is of critical impor-tance. Thus, the restoration of suppressor gene function is a major goal of gene therapy for cancer. The deletion, inactiva-tion and mutation of tumor suppressor gene p16 are quite common in many tumors, especially in glioma. This study was designed to investigate the effect of insertion of p16 gene in suppressing the growth of a human glioma cell line. METH-ODS: The p16 gene was transfected into human glioma cell line TJ899 through adenoviral vector(Ad5). The success of transfection was confirmed by testing the p16 gene expression using immunohistochemical method. The proliferation of the cell line was determined by MTT and colony formation assay. RESULTS: The p16 gene expression was observed in TJ899 cell line that was transfected with recombinant adenovirus (Ad - p16) . The proliferation rate of the Ad-p16 TJ899 cells was inhibited by 93% as compared with that of the Ad-LacZ transfected cells (control). Colony formation in Ad-p16 transfected cells was significantly reduced as compared with Ad LacZ tranfected cells. CONCLUSION: The expression of exogenous p16 inserted in a human glioma cell line, using adenovirus as vector, can inhibit growth and colony formation of the infected glioma cells.