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为了研究端粒酶反义RNA对膀胱癌T24细胞恶性表型的抑制及促进其凋亡的作用,采用脂质体转染法将能转录出端粒酶反义RNA质粒导入膀胱癌T24细胞,应用PCR-ELISA法测定转染后T24细胞的端粒酶活性;光镜、电镜、MTT法及流式细胞检测(FCM)等方法观察端粒酶反义RNA对T24细胞的生长及凋亡的影响。结果显示,端粒酶反义RNA能显著抑制T24细胞的端粒酶活性,使其生长受抑制。形态学观察转染后T24细胞出现典型的凋亡现象。FCM检测发现细胞周期G1期前出现亚2倍体峰——凋亡峰。结论:转染端粒酶反义RNA能抑制膀胱癌T24细胞的恶性表型,并促进其凋亡,从而为以端粒酶作为膀胱肿瘤基因治疗的新靶点提供实验依据。
To study the inhibitory effect of telomerase antisense RNA on the malignant phenotype of bladder cancer T24 cells and to promote its apoptosis, antisense RNA plasmids capable of transcribing telomerase were transfected into bladder cancer T24 cells by lipofection method, The telomerase activity of transfected T24 cells was detected by PCR-ELISA. The growth and apoptosis of T24 cells were observed by light microscopy, electron microscopy, MTT and flow cytometry (FCM) influences. The results showed that telomerase antisense RNA could significantly inhibit the telomerase activity of T24 cells and inhibit its growth. Morphological observation T24 cells after transfection typical apoptotic phenomenon. The sub-2ploid-apoptotic peak appeared before the G1 phase of the cell cycle by FCM. CONCLUSION: Transfection of telomerase antisense RNA can inhibit the malignant phenotype of bladder cancer T24 cells and promote its apoptosis, which provides an experimental basis for using telomerase as a new target for bladder cancer gene therapy.