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目的探讨急性早幼粒白血病基因(PML)对于膀胱肿瘤细胞株UM-UG2的体外增殖抑制作用及其诱导凋亡的作用。方法应用脂质体Lipofectamine2000建立稳定可诱导表达的膀胱肿瘤细胞株UM-UG-2,Western blot检测PML蛋白的表达情况；噻唑蓝(MTT)比色法检测肿瘤生长情况；Giemsa染色、TUNEL和DNA ladder法检测肿瘤的凋亡情况。结果经100/μmol/L的ZnSO_4诱导表达后,与对照组比较PML组细胞生长明显受到抑制并且细胞周期停滞在G_1期。Giemsa染色提示过表达PML蛋白的细胞核浓染,细胞变圆,DNA ladder电泳提示转染PML细胞细胞在诱导表达后24h出现梯状凋亡特征的电泳条带,TUNEL显示转染PML细胞细胞核内蓝褐色颗粒,对照组未见明显凋亡特征性改变。结论建立了稳定的可诱导表达PML的膀胱肿瘤细胞株。同时,过表达PML可以抑制膀胱肿瘤细胞株UM-UC-2的体外增殖作用,诱导膀胱肿瘤细胞凋亡。
Objective To investigate the inhibitory effect of acute promyelocytic leukemia (PML) gene on the proliferation of bladder cancer cell line UM-UG2 and the induction of apoptosis. Methods The stable and inducible expression of bladder cancer cell line UM-UG-2 was established by Lipofectamine 2000. The expression of PML protein was detected by Western blot. The growth of tumor was detected by MTT assay. The expression of Giemsa staining, TUNEL and DNA ladder method to detect tumor apoptosis. Results After the cells were induced with 100 / μmol / L ZnSO_4, the cell growth in PML group was significantly inhibited and the cell cycle arrest was in G_1 phase compared with the control group. Giemsa staining indicated that the PML protein was overexpressed in nuclei and the cells became round. DNA ladder electrophoresis suggested that the ladder-like apoptotic characteristics appeared in transfected PML cells 24h after transfection, TUNEL showed that the translocation of PML Brown particles, the control group showed no significant changes in apoptosis. Conclusion A stable bladder cancer cell line that can induce the expression of PML was established. At the same time, Overexpression of PML can inhibit the proliferation of bladder cancer cell line UM-UC-2 in vitro and induce the apoptosis of bladder tumor cells.