三七总皂甙对人肝癌细胞的抑制作用

Inhibitory effects on human hepatocarcinoma cells with panax notoginseng saponins

目的:观察三七总皂甙对人肝癌细胞SMMC-7721增殖、凋亡及缝隙连接细胞间通讯的影响。方法:实验于2005-08/2006-02在江西省分子医学重点实验室完成。①人肝癌细胞株SMMC-7721由江西省医学科学研究所提供,三七总皂甙注射剂(从五加科人参属多年生草本植物三七中提取的有效成分,昆明制药集团股份有限公司)。②以二苯基四氮唑溴盐比色法观察三七总皂甙对SMMC-7721细胞增殖的影响,分为三七总皂甙25,50,100,200,400,800mg/L组,设不加三七总皂甙为空白对照。③用流式细胞仪检测细胞周期时相分布及细胞凋亡率,并采用划痕标记/染料示踪技术观察三七总皂甙对SMMC-7721细胞的缝隙连接细胞间通讯功能的影响。结果:①三七总皂甙对SMMC-7721细胞增殖的抑制作用:随着三七总皂甙浓度增大、作用时间延长,其对SMMC-7721细胞增殖的抑制作用逐渐增强,呈明显的浓度-时间依赖关系,至800mg/L则有细胞毒性。②三七总皂甙对SMMC-7721细胞周期时相分布和细胞凋亡率的影响:不同浓度的三七总皂甙处理细胞72h后,与空白对照组相比,G0/G1期细胞明显增多,S期和G2/M期细胞减少,且三七总皂甙浓度越高,该作用越强,其中三七总皂甙400mg/L组72h后G0/G1期细胞高达87.42%;同时三七总皂甙还能诱导SMMC-7721细胞凋亡,并呈明显的剂量效应正比关系。③三七总皂甙对SMMC-7721细胞的缝隙连接细胞间通讯功能的影响:三七总皂甙可上调或恢复SMMC-7721细胞的缝隙连接细胞间通讯的作用,400mg/L三七总皂甙处理72h后,荧光染料传输的范围可达4～5层细胞。结论:三七总皂甙可抑制SMMC-7721细胞增殖、促进细胞凋亡,使细胞生长阻滞于G0/G1期,同时上调或恢复细胞的缝隙连接细胞间通讯功能,其抗肿瘤作用可能与此有关。

AIM：To investigate the effects of panax notoginseng sapenins （PNS） on proliferation, apoptosis and gap junctional intercelluar communication （GJIC）of SMMC-7721 human hepatocarcinoma cells. METHODS：The experiment was conducted in the Jiangxi Key Laboratory of Molecular Medicine between August 2005 and February 2006.①Human hepatocarcinoma cells SMMC-7721 was provided by Jiangxi Institute of Medical Sciences, PNS injection （effective components extracted from perennial herb notoginseng, radix, manufactured by Kunming Pharmaceutical Co., Ltd.）.②MTT assay was used to study the effects of PNS on cell proliferation of SMMC-7721 cells. Cells were divided into 25, 50, 100, 200, 400, 800 mg/L group, and non-PNS group was taken as the blank control group.③Cell apeptosis and distribution of cell cycles were examined with flow cytometry. Meanwhile,effect on GJIC of SMMC-7721 cells was observed with scrape labeling/dye tranSfer（SL/DT）technique. RESULTS： ①Inhibitory effect of PNS on proliferation of SMMC-7721 cells： As the concentration of PNS increasing and the time prolonging, the inhibitory effect on proliferation of SMMC-7721 cells was gradually reinforced, which was in a concentration-time dependent manner, and there was cytotoxicity at 800 mg/L.②Effects of PNS on distribution and apeptosis of SMMC-7721 in cell cycle： compared with blank control group at 72 hours after treated with PNS at different concentration, the cells at G0/G1 phase were significantly increased, while cells at S phase and G2/M phase obviously decreased. Moreover, the higher the concentration of PNS was, the stronger the effect was. 72 hours later, cells in the 400 mg/L PNS group at Go/G1 phase were as high as 87.42%. Meanwhile, PNS could induce the apeptosis of SMMC-7721 dells and the effects of PNS were in a dose-dependent manner.③Effects of PNS GJIC of SMMC-7721 cells： PNS could also up-regulate or recover GJIC of SMMC-7721 cells. After treated with 400 mg/L PNS for 72 hours ,the transferring range of LY fluorescence could reach 4-5 rows of cells from the scraped line. CONCLUSION：PNS can inhibit the proliferation and promote apeptosis of SMMC-7721 cells as well as arrest the cell growth at G0/G1 phase of cell cycle. Furthermore,PNS can up-regulate or recover GJIC function of SMMC-7721. cells, which maybe correlate with the anti-tumor effect.