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辣椒素介导活性氧产生及线粒体膜电位去极化抑制膀胱癌细胞增殖的实验研究

发表时间:2017-03-15  浏览量:1920  下载量:298
全部作者: 杨中华,王行环
作者单位: 武汉大学中南医院泌尿外科
摘 要: 目的:探讨辣椒素(capsaicin,CAP)对膀胱癌T24细胞增殖及其荷瘤裸鼠体内移植肿瘤生长的影响。方法:逆转录PCR(reverse transcription-PCR,RT-PCR)及Western blotting检测T24细胞瞬时受体电位香草素1型(transient receptor potential vanilloid type-1,TRPV1)的表达;MTT和流式细胞术检测CAP对T24细胞活力及增殖的影响;JC-1染料法检测CAP对T24细胞活性氧(reactive oxygen species,ROS)生成及线粒体膜电位的影响;荷瘤裸鼠模型评估CAP(5 mg/kg体重,皮下注射)对膀胱癌体内肿瘤生长的影响。结果:T24细胞表达有功能的TRPV1通道。CAP可降低T24细胞活力,并呈剂量依赖性。CAP可激活TRPV1通道介导钙离子内流而诱导细胞死亡,而非凋亡,其机制在于诱导ROS产生和线粒体膜电位去极化。动物实验显示CAP显著减缓T24膀胱癌移植瘤的生长[(567.02±43.94)mm3 vs(661.80±62.03)mm3,P<0.01]。结论:CAP激活TRPV1通道,引起细胞内钙超负荷,诱导ROS生成和线粒体膜电位去极化,进而引起T24细胞死亡。CAP有望为膀胱癌局部及全身化疗提供新思路。
关 键 词: 泌尿外科学;膀胱癌;钙通道;辣椒素;活性氧
Title: Study on capsaicin mediating cell proliferation in bladder cancer through reactive oxygen species production and depolarization of mitochondrial membrane potential
Author: YANG Zhonghua, WANG Xinghuan
Organization: Department of Urology, Zhongnan Hospital of Wuhan University
Abstract: Objective: To investigate the effects of capsaicin (CAP) on cell proliferation of bladder cancer T24 cells in vitro as well as on xenografts in nude mice in vivo. Methods: Reverse transcription-PCR (RT-PCR) and Western blotting were used to assay the expression of transient receptor potential vanilloid type-1 (TRPV1) in bladder cancer T24 cells. Then T24 cells were assessed for cell viability and apoptosis by MTT assay and flow cytometry analysis after incubation with different concentrations of CAP. To uncover the mechanism by which CAP affected the viability of T24 cell, intracellular production of reactive oxygen species (ROS) and mitochondrial membrane potential were assessed by JC-1 stain. To study the in vivo effects of CAP, T24 cells were grown as xenografts in nude mice and CAP (5 mg/kg b.w.) was given by subcutaneous injection in nude mice with bladder tumor. Results: The functional TRPV1 channel could be expressed in T24 cells. CAP decreased the viability of T24 cells in a dose-dependent manner. CAP induced ROS production and mitochondrial membrane depolarization through activate the TRPV1 channel to mediate the calcium inflow, hence induced cell death, not apoptosis. In vivo experiment showed that CAP significantly slowed the growth of T24 bladder cancer xenografts as measured by size [(567.02±43.94) mm3 vs (661.80±62.03) mm3, P<0.01]. Conclusion: CAP mediates cell death in T24 cells through calcium entry-dependent ROS production and mitochondrial depolarization and may have a role in the management of bladder, which is expected to provide new ideas for local and systemic chemotherapy of bladder cancer.
Key words: urology; bladder cancer; calcium channels; capsaicin; reactive oxygen species
发表期数: 2017年3月第5期
引用格式: 杨中华,王行环. 辣椒素介导活性氧产生及线粒体膜电位去极化抑制膀胱癌细胞增殖的实验研究[J]. 中国科技论文在线精品论文,2017,10(5):502-508.
 
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