CAJ | 학술논문

目的了解辣椒素对膀胱癌RT4细胞生长的影响及可能机制.方法采用细胞计数kit-8(CCK-8)试剂盒观察辣椒素(50、100、150、200、250μmol/L)对膀胱癌RT4细胞生长的影响,以辣椒素(0 μmol/L)为对照组;流式细胞术检测细胞周期和凋亡;逆转录-聚合酶链式反应(RT-PCR)和免疫荧光检测瞬时受体电位香草酸亚型1(TRPV1)的表达;Western印迹检测细胞周期蛋白P53、P21、细胞周期依赖性激酶(CDK)2表达水平.结果 100 μmol/L辣椒素明显抑制RT4细胞生长,细胞成活率为82.0%±6.2%,低于对照组(100.0%±12.4%,P=0.036),且生长抑制呈剂量依赖性,250μmol/L时细胞成活率仅为7.8%±2.9%(P=0.000).辣椒素能诱导RT4细胞G0/G1期阻滞,同样呈剂量依赖性,对照组G0/G1期细胞比例为37.4%±5.6%,而250 μmol/L时达到72.4%±5.3%(P=0.000).RT4细胞表达TRPV1受体mRNA和蛋白.与对照组比较,辣椒素处理后48 h,P53、P21表达上调,而CDK2表达下调.结论辣椒素可以通过TRPV1受体调节P53、P21、CDK2表达以诱导人膀胱癌RT4细胞G0/G1期阻滞而抑制其生长.
목적 료해랄초소대방광암RT4세포생장적영향급가능궤제.방법 채용세포계수kit-8(CCK-8)시제합관찰랄초소(50、100、150、200、250μmol/L)대방광암RT4세포생장적영향,이랄초소(0 μmol/L)위대조조;류식세포술검측세포주기화조망;역전록-취합매련식반응(RT-PCR)화면역형광검측순시수체전위향초산아형1(TRPV1)적표체;Western인적검측세포주기단백P53、P21、세포주기의뢰성격매(CDK)2표체수평.결과 100 μmol/L랄초소명현억제RT4세포생장,세포성활솔위82.0%±6.2%,저우대조조(100.0%±12.4%,P=0.036),차생장억제정제량의뢰성,250μmol/L시세포성활솔부위7.8%±2.9%(P=0.000).랄초소능유도RT4세포G0/G1기조체,동양정제량의뢰성,대조조G0/G1기세포비례위37.4%±5.6%,이250 μmol/L시체도72.4%±5.3%(P=0.000).RT4세포표체TRPV1수체mRNA화단백.여대조조비교,랄초소처리후48 h,P53、P21표체상조,이CDK2표체하조.결론 랄초소가이통과TRPV1수체조절P53、P21、CDK2표체이유도인방광암RT4세포G0/G1기조체이억제기생장.
Objective To study the effects of capsaicin on the growth of bladder cancer RT4 cell and its potential mechanism. Methods Cell counting kit-8 (CCK-8) assay and flow cytometry were employed to observe the effects of capsaicin (50, 100, 150,200, 250 μmol/L) on cell growth, cell cycle and apoptosis. Capsaicin (0μmol/L) was used as a control. The effects of mRNA and protein of transient receptor potential cation channel subfamily V 1 (TRPV1) on RT4 cells were tested by RT-PCR and immunofluoresence respectively. And the expressions of cell cycle protein P53, P21, CDK2 were detected by Western blot after the treatment of capsaicin. Results 100 μmol/L capsaicin significantly decreased the viability of RT4 cell [82.0% ± 6. 2% vs 100. 0% ± 12.4% ( control ), P = 0. 036] while the cell viability was 7.8% ±2. 9% at 250 μmol/L (P =0. 000). It was in a dose-dependent manner. On the other hand,capsaicin induced the cell cycle arrest of bladder cancer RT4 cells G0/G1 phase in a dose-dependent way.The cell proportion of G0/G1 phase in the control was 37.4% ±5.6% ,however, it was 72.4% ±5. 3% at 250 μ mol/L (P =0. 000). It was showed that TRPV1 mRNA and protein were expressed in RT4 cells.After a 48-hour treatment with capsaicin, the expressions of P53 and P21 were up-regulated in contrary to the expression of CDK2. Conclusion Capsaicin induces the cell cycle arrest of bladder cancer RT4 cells G0/G1 phase and growth inhibition via TRPV1 receptor by modulating the expression of P53, P21 and CDK2.