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目的观察异丹叶大黄素(ISO)对膀胱癌细胞增殖及X染色体连锁凋亡抑制蛋白(XIAP)的影响,并观察异丹叶大黄素对化疗药物多西他赛诱导膀胱癌细胞凋亡及其细胞毒性的增敏作用。方法以异丹叶大黄素作用于人源性膀胱癌T24T细胞,以ATPase法检测异丹叶大黄素对膀胱癌细胞的细胞毒性,并以软琼脂集落形成实验,检测异丹叶大黄素对T24T增殖能力的影响。以逆转录-聚合酶链反应和蛋白质印迹法检测异丹叶大黄素对膀胱癌细胞X染色体连锁凋亡抑制蛋白基因表达的影响。以ATPase法、倒置显微镜下以及流式细胞仪分别观察检测异丹叶大黄素对多西他赛诱导的膀胱癌细胞凋亡和细胞毒性的影响。结果异丹叶大黄素可显著抑制膀胱癌细胞的增殖和集落形成,其IC50约为(54.5±3.6)μmol·L-1。经蛋白印迹法和逆转录-聚合酶链反应法证实,异丹叶大黄素在作用于人源性膀胱癌T24T细胞后,其X染色体连锁凋亡抑制蛋白基因在蛋白和mRNA水平均显著性降低。经多西他赛处理T24T细胞后,IC50为(8.78±1.32)nmol·L-1;而联用异丹叶大黄素后,IC50仅为(1.02±0.38)nmol·L-1,两者之间有显著性差异(P<0.01)。经流式细胞仪检测,T24T细胞接受2.5 nmol·L-1多西他赛组凋亡率(21.07±2.79)%显著低于联用异丹叶大黄素治疗组的凋亡率(49.59±5.67)%(P<0.01)。结论异丹叶大黄素可抑制膀胱癌细胞增殖,并可通过下调X染色体连锁凋亡抑制蛋白基因表达,显著增强多西他赛诱导的膀胱癌细胞凋亡,并增强细胞毒性。
Objective To observe the effect of iso-dentin emodin (ISO) on the proliferation of bladder cancer cells and the inhibitor of X-linked inhibitor of apoptosis (XIAP), and to observe the apoptosis of bladder cancer cells induced by docetaxel Its cytotoxic sensitization. Methods The human bladder cancer T24T cells were treated with different doses of emodin and the cytotoxicity of different doses of emodin on bladder cancer cells was detected by ATPase. The soft agar colony formation assay was used to detect the effect of different doses of emodin on T24T cells Effects of proliferation. Effects of Isoelandin Emodin on X - linked Inhibitory Protein Gene Expression of Bladder Cancer Cells by Reverse Transcription - polymerase Chain Reaction and Western Blot. The effects of isoflurane on apoptosis and cytotoxicity of docetaxel-induced bladder cancer cells were detected by ATPase, inverted microscope and flow cytometry respectively. Results The isodenasal emodin significantly inhibited the proliferation and colony formation of bladder cancer cells with IC50 of (54.5 ± 3.6) μmol·L-1. Western blotting and reverse transcription - polymerase chain reaction confirmed that isofan emodin in human bladder cancer T24T cells, the X - linked inhibitor of apoptosis protein in the protein and mRNA levels were significantly lower . The IC50 was (8.78 ± 1.32) nmol·L-1 after treated with docetaxel, while the IC50 was only (1.02 ± 0.38) nmol·L-1 when combined with isoflurane, There was a significant difference (P <0.01). Flow cytometry showed that the apoptosis rate of T24T cells receiving docetaxel 2.5 nmol·L -1 (21.07 ± 2.79)% was significantly lower than that of the cells treated with isoflurane emodin (49.59 ± 5.67% )% (P <0.01). CONCLUSION: Isodenas emodin inhibits the proliferation of bladder cancer cells and significantly enhances the apoptosis of bladder cancer cells induced by docetaxel and enhances cytotoxicity by down-regulating the expression of X-linked inhibitor of apoptosis protein.