摘要
目的探讨电压门控钠离子通道Nav1.5表达对结直肠癌(Colorectal cancer,CRC)多药耐药细胞迁移及侵袭力的影响及可能机制。方法通过浓度递增法诱导人CRC亲本细胞(SW620)以构建耐奥沙利铂细胞(SW620/OxR)。采用Transwell小室及CCK-8法比较SW620细胞和SW620/OxR细胞侵袭、迁移及增殖能力差异,蛋白质免疫印迹法(Western blot,WB)及全细胞膜片钳技术检测SW620细胞和SW620/OxR细胞Nav1.5、P-糖蛋白(P-glycoprotein,P-pg)表达水平及细胞膜Na+电流水平,观察钠离子通道抑制剂河豚毒素(Tetrodotoxin,TTX)对SW620细胞和SW620/OxR细胞上述指标的影响。结果与SW620细胞相比,SW620/OxR细胞的侵袭及迁移力显著增加(P<0.05),而增殖力无显著变化(P>0.05),Nav1.5蛋白和P-gp蛋白表达水平显著上调(P<0.05),Na+电流水平也显著升高(P<0.05);TTX能显著降低SW620细胞和SW620/OxR细胞的迁移力和侵袭力(P<0.05),且降低Nav1.5蛋白表达水平(P<0.05);而TTX对SW620细胞和SW620/OxR细胞的增殖力均无影响(P>0.05)。结论Nav1.5在结直肠癌SW620耐药细胞中呈高表达,且伴随Na+电流水平和耐药细胞的迁移力、侵袭力显著增加;其抑制剂TTX能显著降低CRC耐药细胞的迁移及侵袭力,这可能与Nav1.5蛋白表达下调有关,提示Nav1.5激活可能参与结直肠癌的多药耐药(Multi-drug Resistance,MDR)过程。
Objective To explore the effect and possible mechanism of voltage-gated sodium ion channel Nav1.5 protein expression on the migration and invasiveness of colorectal cancer(CRC)multidrug resistant cells.Methods Human CRC parental cells(SW620)were induced by increasing concentration method to construct oxaliplatin-resistant cells(SW620/OxR).Transwell chamber and CCK-8 method were used to compare the difference of invasion,migration and proliferation ability in SW620 cells and SW620/OxR cells.Western blotting(WB)and whole-cell patch-clamp technique were used to detect Nav1.5,P-glycoprotein(P-pg)expression level and cell membrane Na+current level in SW620 cells and SW620/OxR cells.The effect of the sodium channel inhibitor Tetrodotoxin(TTX)on SW620 cells and SW620/OxR cells were observed.Results Compared with SW620 cells,the invasion and migration ability of SW620/OxR cells were significantly increased(all P<0.05),but the proliferation was not significantly changed(P>0.05).The expression level of Nav1.5 and P-gp protein were significant up-regulated(all P<0.05),Na+current level also increased significantly(P<0.05).TTX significantly reduced the migration and invasiveness of SW620 cells and SW620/OxR cells(P<0.05),and decreased Nav1.5 protein expression level(all P<0.05);while TTX had no effect on the proliferation of SW620 cells and SW620/OxR cells(P>0.05).Conclusion Nav1.5 was highly expressed in colorectal cancer SW620 drug-resistant cells,and significantly increased with Na+current level and migration and invasion of drug-resistant cells.Its inhibitor TTX can significantly reduce the migration and invasion of CRC resistant cells,which may be related to the down-regulation of Nav1.5 protein expression,suggesting that Nav1.5 activation may be involved in the multi-drug resistance(MDR)process of colorectal cancer.
作者
胡瑶瑶
汪谢嫒
冯光勇
岳国军
杨明镇
张贵海
Hu Yaoyao;Wang Xieai;Feng Guangyong;Yue Guojun;Yang Mingzhen;Zhang Guihai(Caner Hospital,Afficiated Hospital of Zunyi Medical University,Zunyi Guizhou 563099,China;Department of Oncology,Affiliated Hospital of Xiangnan University,Chenzhou Hunan 423000,China;Shenzhen Yantian District People's Hospital,Shenzhen Guangdong 518000,China;The Second Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563000,China;Department of Intervention,Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563099,China)
出处
《遵义医科大学学报》
2020年第4期425-430,共6页
Journal of Zunyi Medical University
基金
国家自然科学基金资助项目(NO:81260370)
贵州省科技厅联合基金资助项目(NO:黔科合LH[2014]7585)
遵义市科技局基金资助项目(NO:遵市科合社字[2011]25)
遵义市汇川区科技局资助项目(NO:209.001.122.02)。
关键词
结直肠癌
NAV1.5
多药耐药
侵袭
迁移
colorectal cancer
Nav1.5
multi-drug resistance
migration
invasion
