论文部分内容阅读
目的探讨蛋白激酶A(PKA)活性下降对膀胱移行细胞癌细胞株T24增殖和化疗敏感性的影响及其机制。方法用0、15、30μmol/L H-89(PKA活性抑制剂)诱导T24细胞后,用放射免疫法检测H-89作用下T24细胞中PKA活性的变化；用流式细胞仪检测H-89作用下细胞凋亡的变化；用噻唑蓝(MTT)比色法检测丝裂霉素和表阿霉素单独作用下以及与15μmol/L H-89共同作用下T24细胞活力的变化；用Western blot检测15μmol/L H-89作用下T24细胞中bcl-2表达的变化。结果H-89能够使T24细胞中的PKA活性下降；在15μmol/LH-89作用36h后,流式细胞仪检测可见T24细胞凋亡率为57．81%,显著高于对照组4．37%(F=311．35,P＜0．01)。丝裂霉素(MMC)和表阿霉素(Epirubicin)对T24细胞的半数致死量分别为120mg/L和100mg/L；加入15μmol/L H-89后,细胞的存活率分别降为(31．14±4．37)%和(24．91±3．59)%,两组间差异有统计学意义(丝裂霉素组：F=23．94,P＜0．01；表阿霉素组：F=50．64,P＜0．01)。用15μmol/L H-89诱导T24细胞后,bcl-2表达呈下降趋势,并显著低于对照组。结论抑制PKA活性能够使bcl-2的表达下降,从而诱导膀胱癌T24细胞株凋亡,并提高其对化疗药物的敏感性。
Objective To investigate the effect of protein kinase A (PKA) activity on the proliferation and chemosensitivity of human bladder transitional cell line T24 and its mechanism. Methods T24 cells were induced with 0, 15, 30μmol / L H-89 (inhibitor of PKA activity), and the changes of PKA activity in T24 cells treated with H-89 were detected by radioimmunoassay. (MTT) assay was used to detect the cell viability of T24 cells treated with mitomycin and epirubicin alone and with 15μmol / L H-89. Western blot The changes of bcl-2 expression in T24 cells treated with 15μmol / L H-89 were detected. Results After treated with 15μmol / LH-89 for 36h, the apoptotic rate of T24 cells was 57.81%, which was significantly higher than that of the control group (4.37%) by H-89. (F = 311.35, P <0.01). The median lethal doses of MMC and Epirubicin to T24 cells were 120mg / L and 100mg / L, respectively. After addition of 15μmol / L H-89, the cell viability decreased to (31 .14 ± 4.37)% and (24.91 ± 3.59)%, respectively, with significant difference between the two groups (mitomycin group: F = 23.94, P <0.01; Supramolecular group: F = 50.64, P <0.01). After induced by 15μmol / L H-89, the expression of bcl-2 showed a downward trend, which was significantly lower than that of the control group. Conclusion Inhibition of PKA activity can decrease the expression of bcl-2, induce the apoptosis of bladder cancer T24 cell line and increase its sensitivity to chemotherapeutic drugs.