反义端粒酶RNA对人胃癌细胞端粒长度及端粒酶活性调控的研究

 目的　探讨反义端粒酶RNA(hTR)对人胃癌细胞 端粒长度(TRF)及端粒酶活性的调控作 用。方法　应用反义hTR真核表达载体经脂质体介导转染胃癌细胞SGC7901 ，通过分子杂交及端粒重复扩增PCR方法检测hTR表达及端粒、端粒酶活性变化。结 果　经HYR筛选，转染细胞形成稳定克隆，反义hTR表达增强、正义hTR表达下调，平 均TRF缩短、端粒酶活性受抑。结论　反义hTR对胃癌细胞TRF及端粒酶活性 的调控可能是通过其对hTR模板的“封闭”而发挥作用的，反义hTR可以作为胃癌基因治疗的 一种新靶点。

Antisense Human Telomerase RNA Mediated Inhibition of Telomere and Telomerase Activity in Human Gastric Cancer Cells

Objectivc To observe effects of antisense human telomerase RNA(hTR) on telomeric length (TRF) and telomerease activity of human gastric cancer cell SGC7901.Methods SGC7901 cell line was transfected with antisense hTR expression vector(pBBS-hTR) by lipofectamine,and expressions of hTR,telomere and telomerase activity in the gene transfected cells were detected by hybridization of nucleic acids and a telomeric repeat amplification protocol(TRAP).Results Clones containing antisense or control plasmids were selected 3 weeks after transfection by HyR selection and either antisense hTR expression had been enhanced or sense hTR expression had been inhibited 72h after transfection by specific stained RNA probe.Telomerase activity and mean TRF length were general low in the gene transfected cells in comparision with those in the control plasmid transfected cells.Conclusion Antisense hTR may either control TRF length by blocking the template region of hTR or serve as a effective target for treating gastric cancer by inhibiting telomerase.