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目的研究芒柄花黄素对膀胱癌细胞的凋亡作用及对β-catenin通路的影响。方法选择不同浓度(20、40、80μmol/L)的芒柄花黄素作用膀胱癌T24、EJ细胞,同时设对照组(芒柄花黄素浓度为0μmol/L),采用CCK8法检测两种细胞被各浓度药物处理0、24、48和72 h后的细胞增殖情况,采用流式细胞术检测两种细胞被各浓度药物处理48 h的细胞凋亡率,Hoechst 33258荧光染色法检测T24细胞被各浓度药物处理48 h后的细胞凋亡情况,RT-PCR检测T24细胞被各浓度药物处理48 h后β-catenin mRNA表达水平。结果 CCK8实验显示芒柄花黄素明显抑制膀胱癌T24、EJ细胞增殖,呈剂量和时间依赖性(P<0.05),并且药物对T24细胞的抑制增殖效果比EJ细胞高(P<0.05);流式细胞术显示T24细胞的对照组及20、40、80μmol/L芒柄花黄素组48 h后凋亡率分别为(1.75±0.71)%、(5.21±1.02)%、(17.78±1.67)%、(30.48±2.51)%,而EJ细胞凋亡率分别为(1.95±1.21)%、(4.84±1.35)%、(14.87±2.59)%、(21.84±2.64)%,与对照组比较,两种细胞的40、80μmol/L芒柄花黄素组凋亡率显著增高(P<0.05),并且芒柄花黄素对T24细胞的凋亡影响比对EJ细胞更明显(P<0.05);Hoechst33258染色显示T24细胞随药物浓度的升高,凋亡明显增多;RT-PCR结果显示T24细胞中β-catenin的mRNA表达水平随着药物浓度升高而降低(P<0.05)。结论芒柄花黄素能抑制膀胱癌T24细胞和EJ细胞增殖、诱导其凋亡,其机制可能与抑制β-catenin通路有关。
Aim To study the apoptosis of bladder cancer cells and the effect on the β-catenin pathway of mangiferin. Methods Toxoplasmacetin (20,40 and 80μmol / L) was used to treat T24 and EJ cells in bladder cancer. At the same time, the control group (0μmol / L of mangrove flavonoids) was treated with CCK8 Cells were treated with various concentrations of drugs for 0,24,48 and 72 h after the proliferation of cells by flow cytometry were treated with various concentrations of drugs for 48 h apoptosis rate, Hoechst 33258 fluorescence staining T24 cells The apoptotic cells were treated with various concentrations of drugs for 48 h. The expression of β-catenin mRNA in T24 cells treated with various concentrations of drugs for 48 h was detected by RT-PCR. Results The results of CCK8 showed that the apoptosis of bladder cancer T24 and EJ cells was significantly inhibited by monascus yellow in dose and time dependent manner (P <0.05), and the inhibitory effect of drug on T24 cells was higher than that of EJ cells (P <0.05). Flow cytometry showed that the apoptotic rates of T24 cells in control group and 20, 40, 80μmol / L of manganic stem cell were (1.75 ± 0.71)%, (5.21 ± 1.02)%, (17.78 ± 1.67 ) And (30.48 ± 2.51)%, respectively. The rates of EJ cell apoptosis were (1.95 ± 1.21)%, (4.84 ± 1.35)%, (14.87 ± 2.59)% and (21.84 ± 2.64)%, respectively. Compared with the control group (P <0.05), and the apoptotic rate of T24 cells was more obvious than that of EJ cells (P <0.05) ); Hoechst33258 staining showed that the T24 cells with the drug concentration increased significantly increased apoptosis; RT-PCR results showed that the mRNA expression ofβ-catenin in T24 cells decreased with increasing drug concentration (P <0.05). Conclusion It is suggested that canonon may inhibit the proliferation and induce the apoptosis of T24 and EJ cells, which may be related to the inhibition of β-catenin pathway.