三七皂苷R1通过细胞缝隙连接对顺铂细胞毒性作用的影响

Panax notoginseng saponins R1 enhances the cytotoxicity of cisplatin via gap junction intercellular communication

目的:探讨三七皂苷R1对稳定表达连接蛋白26、32的HeLa细胞的细胞缝隙连接(gap junction,GJ)功能的作用及其对顺铂细胞毒性的影响。方法:磺基罗丹明B法观察三七皂苷R1作用不同时间以后对HeLa细胞增殖的抑制作用,分为三七皂苷R1 10、25、50、100、200μmol/L组,以不加三七皂苷R1组为空白对照组。计算药物作用24、48、72 h各组对HeLa细胞的生长抑制率;划痕标记/染料示踪技术检测三七皂苷R1作用48 h荧光黄在细胞之间传递的距离来评价其对GJ功能的影响;标准细胞集落形成分析法观察R1与顺铂合用以后对细胞集落形成的影响,以顺铂细胞集落形成率的降低作为顺铂的细胞毒性的指标。结果:三七皂苷R1对HeLa细胞增殖具有一定的抑制作用;R1具有上调GJ功能的作用;有GJ时R1与顺铂合用后可明显增强顺铂的细胞毒性作用,在无GJ时这种作用则不明显。结论:三七皂苷R1可通过增强细胞之间的GJ功能来增强顺铂的细胞毒性作用。

Objective：To investigate the effect of panax notoginseng saponins R1 on gap junction（GJ） function in HeLa cells transfected with connexin 32 and connexin 26 plasmid and the cytotoxity of cisplatin, nethods：Sulforhodamine B assay was used to determine the inhibitory effect of panax notoginseng saponin R1 on HeLa cells growth for different times. Cells were divided into panax notoginseng saponins R1 10,25,50,100, and 200 μmol/L groups, and non-R1 group was taken as the blank control group. After 24,48, and 72 hours the inhibition rate of HeLa ceils growth was evaluated ; the GJ function of HeLa cells treated with R1 for 48 hours was detected by SL/DT（ scrape-loading dye transfer） ,and distance between cells of lucifer yellow was introduced to reflect the GJ function. The cytotoxity of cisplatin combined with R1 was determined by Standard colony-forming assay, and ＂Surviving Fraction＂ as index was used to evaluate the cytotoxicity of cisplatin. Results： Panax notoginseng saponins R1 suppressed the growth of HeLa cells; R1 up- regulated the GJ of HeLa cells. In the presence of GJ, combination of R1 and cisplatin significantly enhance the cytotoxicity of cisplatin compared with cisplatin treatment group, but in absence of GJ, there was no significant difference between these two groups. Conclusions：R1 enhances the cytotoxicity of cisplatin through enhancement of GJ function.