缝隙连接蛋白43组成的缝隙连接对依托泊苷抗肿瘤作用的影响  

The role of gap junctions composed of connexin 43 on the anti-tumor effect induced by etoposide

作　　者：汪灵芝[1] 彭建新[2] 陶亮[3] 黄焕森[1] 

机构地区：[1]广州医科大学附属第二医院麻醉科,广东广州510260 [2]广东省中医院肝胆外科,广东广州510120 [3]中山大学中山医学院药理学,广东广州510080

出　　处：《中国癌症杂志》2015年第1期1-5,共5页China Oncology

基　　金：国家自然科学基金资助项目(81401017);广州市卫生局西医引导项目(20141A011083)

摘　　要：背景与目的:缝隙连接能够增强化疗药物的细胞毒性,但缝隙连接蛋白43(connexin 43,Cx43)组成的缝隙连接对依托泊苷抗肿瘤作用的影响尚不清楚。因此,本研究拟观察睾丸癌细胞中Cx43对依托泊苷抗肿瘤作用的影响。方法:采用细胞接种荧光示踪实验检测睾丸癌细胞由Cx43形成的缝隙连接通讯功能,同时检测不同的药物(18-α-甘草次酸及维甲酸)和Cx43干扰RNA对缝隙连接功能的影响;采用标准细胞集落形成分析法检测睾丸癌细胞中由Cx43形成的缝隙连接通讯功能改变后,对依托泊苷抑制肿瘤细胞生长作用的影响。结果:荧光示踪实验显示18-α-甘草次酸可显著降低缝隙连接通讯功能,而维甲酸则可增强缝隙连接功能;Cx43干扰RNA能够显著抑制缝隙连接功能。标准细胞集落形成实验结果表明,在缝隙连接形成的睾丸癌细胞中,18-α-甘草次酸显著降低依托泊苷对睾丸癌细胞生长的抑制作用,而维甲酸则增强依托泊苷对睾丸癌细胞生长的抑制作用;应用Cx43干扰RNA能够显著降低依托泊苷对睾丸癌细胞生长的抑制作用。结论:睾丸癌细胞中Cx43形成的缝隙连接通讯功能降低能够减弱依托泊苷的抗肿瘤作用;而缝隙连接通讯功能增强能够增强其抗肿瘤作用。Background and purpose： Gap junctions（GJ） could enhance cytotoxicity induced by chemotherapeutic agents. However, whether or not GJ composed of connexin 43 （Cx43） could increase etoposide cytotoxicity remains unclear. The aim of this study was to explore the effect of GJ composed of Cx43 on etoposide cytotoxicity in testicular cancer cells. Methods： Eighteen-glycyrrhetinic acid and siRNA were used to inhibit GJ function. Retinoid acid was used to enhance GJ function.“Parachute”dye-coupling assay was used to examine dye spread through GJ composed of Cx43 in MLTC-1 cells. “Standard colony-forming assay” was used to examine cell survivals of MLTC-1 cells treated with etoposide. Results： Assayed by “parachute” dye-coupling assay, the dye spread through GJ in MLTC-1 cells was significantly decreased by 18-glycyrrhetinic acid however increased by retinoid acid. Cx43 expression and GJ function in MLTC-1 cells were inhibited by Cx43-siRNA. Results from “standard colony-forming assay” showed that etoposide cytotoxicity was decreased by 18-glycyrrhetinic acid and siRNA, however enhanced by GJ function enhancer retinoid acid. Conclusion：The function inhibition of Cx43 composed GJ in MLTC-1 cells could decrease etoposide cytotoxicity. The enhancement of GJ composed of Cx43 in MLTC-1 could increase etoposide cytotoxicity.

关 键 词：缝隙连接蛋白43 依托泊苷 缝隙连接 

分 类 号：R737.21[医药卫生—肿瘤]