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作 者:陈 琰[1] 董绍忠[1] 王秋枫 张庆诗[1] 杨连甲[1]
出 处:《口腔颌面外科杂志》2002年第1期29-33,共5页Journal of Oral and Maxillofacial Surgery
摘 要:目的 检测SAcc83细胞系端粒酶活性并观察人端粒酶模板hTR封闭后细胞生长行为的改变及与凋亡的关系。方法TRAP-PCR-ELISA法检测SAcc83细胞系端粒酶活性,脂质体介导真核表达载体PBBS212-ahTR转染SAcc83,PCNA免疫组化染色以证实其增殖活性改变,TUNEL法及琼脂糖凝胶电泳检测细胞凋亡。结果转染反义hTR后SAcc83细胞增殖能力明显下降,群体倍增时间延长,转染细胞约30天后死亡。各项凋亡检测均证明细胞凋亡的存在。结论hTR反义基因通过封闭端粒酶模板RNA抑制了端粒酶活性,并可能通过某种途径激活细胞的凋亡程序,这为以端粒酶为靶子治疗恶性肿瘤提供了方向。ve To demonstrate the effect of lipofect AMINE-mediated antisense human telomerase RNA trans-fection on the growth and apoptosis of SAcc83 cell line. Methods The activity of telomerase was examined by TRAP-PCR-ELISA. With the eukaryotic expression vector PBBS212-ahTR,anti-hTR was expressed in Acc83, which was tested by hTR in situ-mRNA hybridization. Immuno-histochemical technique was employed to estimate the proliferation ability of the cells. To determine the occurrence of apoptosis, a DNA fragmentation assay in agarose gel and TUNEL method were performed . Results The anti-hTR transfected cells underwent death after about 30 days, and their proliferating ability were greatly reduced compared to the control groups. Apoptosis was testified by several means. Conclusion Antisense hTR transfection may inhibit the activity of telomerase and induce apoptosis in SAcc83 cell line. Telomerase might become a promising target for cancer gene therapy.
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