辣椒素诱导膀胱癌RT4细胞的细胞周期阻滞

[Induction of cell cycle arrest in bladder cancer RT4 cells by capsaicin].

作者信息

Li Qin, Wang Xing-huan, Yang Zhong-hua, Wang Huai-peng, Yang Zhi-wei, Li Shi-wen, Zheng Xin-min

机构信息

Department of Urology, Zhongnan Hospital Affiliated to Wuhan University, Wuhan 430071, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2010 May 11;90(18):1230-3.

PMID:20646592

Abstract

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 micromol/L) on cell growth, cell cycle and apoptosis. Capsaicin (0 micromol/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 immunofluorescence respectively. And the expressions of cell cycle protein P53, P21, CDK2 were detected by Western blot after the treatment of capsaicin.

RESULTS

100 micromol/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 micromol/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 G(0)/G(1) phase in a dose-dependent way. The cell proportion of G(0)/G(1) phase in the control was 37.4% +/- 5.6%, however, it was 72.4% +/- 5.3% at 250 micromol/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 G(0)/G(1) phase and growth inhibition via TRPV1 receptor by modulating the expression of P53, P21 and CDK2.

摘要

目的

研究辣椒素对膀胱癌RT4细胞生长的影响及其潜在机制。

方法

采用细胞计数试剂盒-8（CCK-8）法和流式细胞术观察辣椒素（50、100、150、200、250微摩尔/升）对细胞生长、细胞周期和凋亡的影响。以辣椒素（0微摩尔/升）作为对照。分别通过RT-PCR和免疫荧光检测瞬时受体电位阳离子通道亚家族V 1（TRPV1）的mRNA和蛋白对RT4细胞的影响。辣椒素处理后，通过蛋白质免疫印迹法检测细胞周期蛋白P53、P21、CDK2的表达。

结果

100微摩尔/升辣椒素显著降低RT4细胞的活力[82.0%±6.2% vs 100.0%±12.4%（对照），P = 0.036]，而在250微摩尔/升时细胞活力为7.8%±2.9%（P = 0.000）。呈剂量依赖性。另一方面，辣椒素以剂量依赖的方式诱导膀胱癌RT4细胞的细胞周期停滞于G(0)/G(1)期。对照组G(0)/G(1)期细胞比例为37.4%±5.6%，而在250微摩尔/升时为72.4%±5.3%（P = 0.000）。结果表明TRPV1 mRNA和蛋白在RT4细胞中表达。辣椒素处理48小时后，P53和P21的表达上调，与CDK2的表达相反。