HERG K+ channel related chemosensitivity to sparfloxacin in colon cancer cells
HERG K+ channel related chemosensitivity to sparfloxacin in colon cancer cells
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DOI：
10.3892/or_00000820
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发表时间：
2010-06-01
期刊：
影响因子：
4.2
通讯作者：
Zhen, Yong-Su
中科院分区：
文献类型：
--
作者：
Gong, Jian-Hua;Liu, Xiu-Jun;Zhen, Yong-Su
钾离子通道对于细胞增殖的调节是必不可少的。据报道，HERG蛋白在包括结肠癌在内的多种人类肿瘤中过表达。本研究旨在探讨抗菌药物司帕沙星（SPFX）对HERG K+通道高表达结肠癌细胞的作用。Western blotting检测HERG和凋亡相关蛋白的表达。采用MTT法检测药物及药物联合的体外细胞毒性。基因转染用于检测hERG相关化疗敏感性的变化。流式细胞仪检测细胞凋亡。明胶酶谱法和Boyden小室法测定SPFX对肿瘤细胞迁移和侵袭能力的影响。在BALB/c小鼠中的鼠结肠癌C26和裸鼠中的人结肠癌HCT 116异种移植物中评估SPFX的体内功效。HERG蛋白在结肠癌C26、HCT 116和HT-29细胞中高表达。SPFX以剂量依赖性方式抑制结肠癌细胞的细胞活力。SPFX诱导结肠癌HCT 116细胞凋亡并抑制其迁移和侵袭。细胞凋亡的增加与caspase-3和Bcl-2蛋白表达的减少有关。用hERG转染的HEK 293细胞和siRNA敲低的HCT 116细胞的研究证实，SPFX对细胞活力的抑制与HERG表达相关。当与5-氟尿嘧啶联合使用时，SPFX在HCT 116和HT-29细胞中显示出协同的抗增殖活性。SPFX对人结肠癌HCT 116移植瘤的生长有明显的抑制作用，并与5-氟尿嘧啶有协同作用。我们的研究结果表明，SPFX可能是一种生化调节剂，在结肠癌化疗药物的治疗。
Potassium channels are essential for the regulation of cell proliferation. As reported, HERG protein is over-expressed in a wide range of human tumors, including colon carcinoma. The aim of this study was to investigate the effects of antibacterial agents sparfloxacin (SPFX), a blocker of HERG channel, on HERG K+ channel highly expressing colon cancer cells. Expression of HERG and apoptosis correlative proteins was examined by Western blotting. The MTT assay was used to detect the cytotoxicity of drugs and drug combination in vitro. Gene transfection was used to examine the changes in herg-related chemosensitivity. Cell apoptosis was analyzed by flow cytometry. The migration and invasion capacity of tumor cells by SPFX was determined by gelatin zymography assay and Boyden chamber. The in vivo efficacy of SPFX was assessed in murine colon carcinoma C26 in BALB/c mice and human colon carcinoma HCT116 xenografts in nude mice. High expression of HERG protein was detected in colon cancer C26, HCT116 and HT-29 cells. The cell viability of the colon cancer cells was inhibited by SPFX in a dose-dependent manner. SPFX induced apoptosis and inhibited migration and invasion of colon cancer HCT116 cells. The increase in apoptosis was associated with a decrease in procaspase-3 and Bcl-2 protein expression. Study with herg-transfected HEK293 cells and siRNA-knock down HCT116 cells confirmed that the cell viability inhibition by SPFX was correlated with HERG expression. When combined with 5-fluorouracil, SPFX showed synergistic anti-proliferation activity in HCT116 and HT-29 cells. Furthermore, SPFX inhibited the growth of human colon carcinoma HCT116 xenografts and showed synergistic effect with 5-fluorouracil in vivo. Our finding suggested that SPFX could be a biochemical modulator in treatment of colon cancer with chemotherapeutic drugs.