全硫代反义寡核苷酸抑制人膀胱癌Ej细胞活性的实验研究

Inhibitory effects of the proliferation of human bladder carcinoma Ej cell by antisense phosphorothioate oligonucleotide

目的研究全硫代反义寡核苷酸(PS-ASODN)对人膀胱癌细胞Ej端粒酶活性及细胞生长的影响,探讨PS-ASODN在膀胱癌治疗中的价值,为临床治疗膀胱癌提供理论依据。方法将PS-ASODN转染作用于人膀胱癌细胞Ej中,分别采用MTT法、流式细胞术、酶联免疫吸附(ELISA)法检测不同浓度PS-ASODN对人膀胱癌细胞Ej的细胞增殖、细胞凋亡和端粒酶活性的作用。结果PS-ASODN作用于人膀胱癌细胞Ej后能显著抑制人膀胱癌细胞Ej的端粒酶活性,抑制癌细胞生长增殖,促进细胞凋亡,而且呈现时间特异性和剂量依赖性,与SODN转染组及空白对照组进行比较,差异显著(P<0.05或0.01)。结论PS-ASODN能抑制人膀胱癌细胞Ej的生长,降低其端粒酶活性并诱导细胞凋亡;抑制端粒酶活性可能成为膀胱癌基因治疗的新靶点。

Objective To study the effects of antisense phosphorothioate oligodeoxy nucleotides （PS-ASODN） on telomerase activity and proliferation of bladder carcinoma Ej cell, and explore the treatment value of PS-ASODN for bladder cancer. Methods PS-ASODN was transfected into bladder cancer cell line Ej by liposomal transfection, Effects of different PS-ASODN density on proliferation activity of cell proliferation, cell withering, activity of telomerase of Ej were examined by methyl thiazolyl tetrazolium （MTT）, flow cytometermethod and ELISA. Results Telomerase activity of Ej cells, transfected with PS-ASODN, was significantly inhibited, compared with that of control groups and nonsense oligonucleotide （SODN） transfected cells （ P〈0. 05 or 0.01） ,and showed no difference between the latter two groups （ P 〉0.05）. Inhibitory effect of PS-ASODN was showed in a dose and time-depended manner. The proliferation of Ej cells transfected with ASODN undergoing apoptosis was statistically lower than that of control groups （ P〈0.05）. Condusion PS-ASODN can inhibit the growth of Ej Cells and decrease telomerase activity, causing the revulsion of cell withering. The results suggested that inhibition of telomerase activity would be a new target to treatment for bladder cancer.