反义hTERT抑制恶性胶质瘤生长的体内外研究

Antisense human telomerase reverse transcriptase inhibits the growth of human glioma cells in vitro and in vivo

目的探讨腺病毒介导的反义hTERT在体内外对恶性胶质瘤细胞生长的影响。方法构建含有hTERT反义序列的腺病毒载体在体内外转染恶性胶质瘤细胞系U251，检测肿瘤细胞生长情况、细胞周期变化、端粒酶活性、hTERT蛋白表达等。结果腺病毒介导的hTERT反义治疗在体外明显抑制肿瘤细胞生长，增殖减慢，凋亡增多，细胞较多聚集在G0／G0期，转染后第6天细胞生存率为46．9％，端粒酶活性和hTERT蛋白表达都明显下降，在体内实验中肿瘤生长明显减慢。结论腺病毒介导的反义hTERT在体内外能明显抑制恶性胶质瘤细胞的生长。

Objective To study inhibitory efficacy of antisense hTERT for malignant gliomas transfected by adenovirus in vitro and in vivo. Methods To construct adenovirus vector containing antisense hTERT and transfect into U251 human malignant glioma cells in vitro and in vivo. U251 cell proliferation in vitro was determined by MTT assay and flow cytometry, tumor growth in vivo was measured the volume of glioma in nude mice. Telomerase activity was detected by TRAP assay ; Expression of hTERT was detected by Western blot and RT-PCR methods. Results After transfection in vitro, the growth of U251 cells was inhibited significantly as a time dependent manner, majority of U251 cells were arrested at G0/G1 stages. At the 6th day, the survival rate was 46.9%. Telomerase activity and hTERT protein expression were inhibited significantly. The growth of tumors in vivo was also effectively inhibited after transfection. Conclusion Growth of malignant glioma cells was effectively inhibited after transfection with antisense hTERT in vitro and in vivo.