大蒜素抑制体外人肝癌细胞的侵袭能力  

Allicin inhibits invasion of human hepatocellular carcinoma cells in vitro

作　　者：钟宁[1] 马亚兵[1] 高海青[1] 张志勉[1] 程梅[1] 由倍安[1] 伊永亮[1] 刘新春[1] 

机构地区：[1]山东大学齐鲁医院消化内科,山东省济南市250012

出　　处：《世界华人消化杂志》2005年第6期743-747,共5页World Chinese Journal of Digestology

基　　金：山东省科技发展计划项目;No.95006~~

摘　　要：目的:肝癌的侵袭与转移是导致其不良预后的重要因素,目前临床缺乏针对肝癌侵袭与转移的有效治疗措施. 本研究通过人肝癌细胞培养,研究大蒜素对体外人肝癌细胞侵袭能力的影响,并在基因水平上探讨其机制. 方法:将人肝BEL-7402细胞传代,取进入指数生长期的细胞随机分为阿霉素组Human(5 mg/L)、大蒜素低、中、高剂量组(25、50、100 mg/L),另设空白对照. 处理后每30 min观察一次细胞转移相关超微结构,8 h 后用流式细胞术检测肿瘤侵袭转移抑制基因nm23-H1 和P2ras表达水平.数据用SAS 8.2软件进行X2检验. 结果:与阿霉素组相比,大蒜素处理后超微结构除凋亡表现外,大多数贴壁细胞胞质回缩,细胞之间的连接减少,空隙加大,细胞之间界限变清晰,表面的丝状微绒毛也明显减少.流式细胞检测显示,nm23-H1表达的荧光强度空白对照组为19.19,5 mg/L.阿霉素组为119.76,25 mg/L大蒜素组为84.28,50mg/L大蒜素组为92.64,100 mg/L大蒜素组为138.08,nm23-H1 表达与大蒜素呈明显剂量效应关系,大蒜素高剂量组比阿霉素组显著增强(138.08 vs 119.76,P<0.05).而P21ras 表达的荧光强度在空白对照组、阿霉素组和低、中、高剂量组分别为2.65%、3.56%、1.55%、3.22%、3.44%,都表现为阴性低表达,各组间没有明显差别. 结论:大蒜素可以抑制体外肝癌细胞的侵袭能力,其机制可能与大蒜素特异性影响BEL-7402细胞袁面的微绒毛和上调肿瘤侵袭与转移抑制基因nm23-H1的表达有关.AIM: To investigate the influence of allicin on the ultramicromorphology of hepatocellular carcinoma cells, and to explore the genetic mechanism by which allicin inhibits the invasion and metastasis of hepatocellular carcinoma. METHODS: Human hepatocellular carcinoma BEL-7402 cells were cultured in the presence of adriamycin (5 mg/L), or allicin at low, medium and high concentrations(25, 50, and 100 mg/L, respectively). Metastasis-related ultrami-croscopic structures were examined 0.5 hour after drug treatment. Tumor invasion and metastasis inhibitory genes nm23-H1 and P21ras were assayed by flow cytometry 8 hours following treatment. The data were analyzed by X2 test with SAS 8.2 software. RESULTS: The growth of BEL-7402 cells was inhibited and apoptosis was induced following allicin treatment. Most of these adherent cells shrinked, with the junctions reduced, the intercellular space widened, and the cell surface microvilli decreased. Although cells treated with adriamycin also showed typical apoptosis morphology, there was no remarkable reduction of the microvilli on cell surface, and the cross junctions of microvilli among cells exhibited no changes either. The fluorescent intensities of the expression of nm23-H1 were 19.19 in negative control group, 119.76 in 5 mg/L adriamycin group, 84.8 in 25 mg/L allicin group, 92.64 50 mg/L allicin group, and 138.08 in 100 mg/L allicin group, respectively. Dose-effect relationship was clearly showed for allicin in the induction of nm23-H1 expression. The fluorescent intensity of 100 mg/L allicin group was remarkably stronger than that of 5 mg/L adriamycin group (138.08 vs 119.76, P<0.05). Negative or low expression of P21ras was observed in all groups, with no difference among them. CONCLUSION: Allicin can inhibit the invasion of hepato-cellular carcinoma cells, which may be caused by its influence on the motor system of hepatocellular carcinoma cells and its upregulation nm23-H1 gene expression.

关 键 词：大蒜素 人肝癌细胞 体外 基因nm23-H1 BEL-7402 P21^RAS 癌细胞侵袭能力 流式细胞术检测 侵袭与转移 流式细胞检测 剂量效应关系 超微结构 转移抑制 肿瘤侵袭 荧光强度 癌细胞培养 阿霉素 X^2检验 高剂量 不良预后 治疗措施 

分 类 号：R735.7[医药卫生—肿瘤] R285.5[医药卫生—临床医学]