中药癌痛克对人肝癌细胞HepG2增殖、凋亡及Rb基因表达的影响  

Influence of Chinese herb aitongke on the proliferation, apoptosis and Rb gene expression of hepatoma carcinoma cells HepG2

作　　者：徐波[1] 朱光辉[1] 夏金堂[1] 李书华[2] 

机构地区：[1]广州市第一人民医院普外科,广东省广州市510180 [2]广州医学院病理教研室,广东省广州市510180

出　　处：《中国组织工程研究与临床康复》2007年第12期2253-2256,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广州市中医药科研资助项目(2005A007)~~

摘　　要：目的:通过观察不同浓度癌痛克对肝癌细胞株HepG2增殖及凋亡的作用,以及在相应状态下细胞内Rb基因表达量的改变,探讨中药癌痛克抗肝癌的可能作用机制。方法:实验于2005-09/2006-03在广州医学院中心实验室完成。取对数生长期的HepG2细胞,使细胞静止于G0/G1期。随机分为癌痛克2,10,50mg/L组和对照组,癌痛克2,10,50mg/L组分别加入对应浓度癌痛克(购自河南省肿瘤研究所,由金蝎、土元、九香虫、大黄、人参、灵芝、黄芪等纯中药组成的粉状制剂,功能:消癌肿、消癌痛、消积水、升白排毒),对照组不加药物,每组设4个复孔。MTT法检测各组细胞24,48,72h增殖率,细胞增殖率=[(A570癌痛克组-A570对照组)/A570对照组]×100%;以流式细胞术检测各组细胞24,48,72h凋亡率;RT-PCR方法检测各组细胞24,48,72h细胞内Rb基因mRNA表达量。结果:①2～50mg/L癌痛克具有明显的增殖抑制作用,癌痛克2,10,50mg/L组在24,48,72h与对照组比较,差异有显著性意义(P<0.05);3组各时点两两之间比较差异有显著性意义(P<0.05);同组各时点之间比较差异有显著性意义(P<0.05)。②2～50mg/L癌痛克组可诱导HepG2细胞凋亡,相同作用时间癌痛克2,10,50mg/L组与对照组比较差异有显著性意义(P<0.001);同一时点各组比较差异有显著性意义(P<0.05);同组不同作用时间点比较差异有显著性意义(P<0.001)。③2～50mg/L癌痛克可上调Rb基因的表达,各浓度组在各时点与对照组比较,差异均有显著性意义(P<0.05);24,48,72h癌痛克2,10,50mg/L组之间比较差异无显著性意义(P>0.05);3组各时点比较差异有显著性意义(P<0.05)。结论:中药癌痛克可通过抑制HepG2细胞增殖或诱导其凋亡,而发挥抗肝癌作用;且其诱导细胞凋亡的机制之一可能为通过增加Rb基因的表达而实现。AIM： To investigate the possible anti-cancer mechanisms of aitongke by observing its impact on the proliferation, apoptosis and the expression of Rb gene in human hepatic cell line HepG2. METHODS： The experiment was conducted in the central laboratory of Guangzhou Medical College from September 2005 to March 2006. HepG2 cells at logarithmic growth phase were cultured until the G0/G1 phase of the cell cycle, then divided into 2, 10, 50 mg/L aitongke groups and control group. The aitongke groups were treated with aitongke at different concentrations （2, 10 and 50 mg/L, purchased from the tumor research institute of Henan, the powder was composed of gold scorpion, eupolyphaga, aspongopus, rhubarb, ginseng, gyrophora, astragali and so on, which could reduce cancer swelling, cancer pain, hydrops, lift white cells and evacuate pus）; the control group was not given any treatment, and 4 replicated wells were set in each group. The proliferating rate at 24, 48 and 72 hours was evaluated by MTT method： cell proliferating rate=[（A570 aitongke group -A570 control group）/A570 control group]×100%; the apoptosis rate and Rb gene mRNA level changes of HepG2 cells at 24, 48 and 72 hours were measured by flow cytometry analysis and RT-PCR method, respectively. RESULTS, （1）Aitongke （2-50 mg/L） could effectively inhibit HepG2 cell proliferation; the differences were significant when compared with the control group （P 〈 0.05）; there were significant differences between any two groups of the three at each time point （P 〈 0.05）; remarkable differences were also found in the same group at different time points （P 〈 0.05）. （2）Aitongke （2-50 mg/L） could effectively inhibit cell apoptosis; the differences were significant when compared with the control group at the same time （P 〈 0.05）; there were significant differences between any two groups of the three at same time point （P 〈 0.05）; remarkable differences were also found in the same group at different time points （P 〈 0.0

关 键 词：癌 肝细胞/病理学 基因.肿瘤抑制 基因 视网膜母细胞瘤/遗传学 抗肿瘤药/药理学 细胞凋亡/药物作用 

分 类 号：R735.7[医药卫生—肿瘤]