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目的探讨芒柄花黄素诱导膀胱癌细胞凋亡的作用及机制。方法采用不同浓度(20、40、80μmol/L)芒柄花黄素处理膀胱癌细胞,采用噻唑蓝法观察芒柄花黄素对膀胱癌T-24和BIU-87细胞的增殖的抑制作用,使用流式细胞术及Hoechst 33258染色检测T-24和BIU-87细胞凋亡率,采用RT-PCR检测T-24细胞Bcl-2 mRNA表达;western blot检测T-24细胞p-p38蛋白激酶、Bcl-2蛋白含量变化。结果与对照组比较,40、80μmol/L芒柄花黄素组T-24和BIU-87细胞增殖率下降、细胞凋亡率明显升高,Hoechst33258染色显示T-24细胞呈凋亡状态;40、80μmol/L芒柄花黄素组T-24细胞Bcl-2 mRNA表达水平[(0.701±0.029)、(0.408±0.036)]均低于对照组,差异均有统计学意义(P<0.01);与对照组比较,20、40、80μmol/L芒柄花黄素组T-24细胞p-p38蛋白激酶活性水平[(0.483±0.026)、(0.569±0.031)、(0.935±0.037)]均升高,Bcl-2蛋白含量[(0.998±0.034)、(0.591±0.030)、(0.426±0.027)]均降低,差异均有统计学意义(P<0.01)。结论芒柄花黄素可诱导膀胱癌T-24和BIU-87细胞凋亡、抑制细胞生长,其机制可能与下调Bcl-2蛋白表达并活化p38蛋白激酶有关。
Objective To investigate the effect and mechanism of apoptosis induced by cinobufotalin in bladder cancer cells. Methods The bladder cancer cells were treated with different concentrations (20, 40, and 80μmol / L) of mangiferin, and the inhibitory effects of monascus on proliferation of bladder cancer T-24 and BIU-87 cells were observed by MTT method. The apoptosis rate of T-24 and BIU-87 cells was detected by flow cytometry and Hoechst 33258 staining. The expression of Bcl-2 mRNA in T-24 cells was detected by RT-PCR. The expressions of p-p38 protein kinase, Bcl-2 protein content changes. Results Compared with the control group, the proliferation rates of T-24 and BIU-87 cells in the 40 and 80μmol / L of manganicusin group were significantly decreased, and the apoptosis rate was significantly increased. The apoptosis of T-24 cells was observed by Hoechst33258 staining. 40 (P <0.01). The expression of Bcl-2 mRNA in T-24 cells treated with 80 μmol / L of manganose was significantly lower than that of the control group [(0.701 ± 0.029), (0.408 ± 0.036) ; Compared with the control group, the p-p38 protein kinase activity of T-24 cells in 20, 40, 80μmol / L manganicin group [(0.483 ± 0.026), (0.569 ± 0.031), (0.935 ± 0.037) (0.998 ± 0.034), (0.591 ± 0.030) and (0.426 ± 0.027), respectively, which were significantly lower than those in control group (P <0.01). CONCLUSION: The expression of Bcl-2 protein and activation of p38 protein kinase may be related to the apoptosis of bladder cancer cell line T-24 and BIU-87 induced by mangiferin.