大蒜素对人视网膜母细胞瘤Rb细胞中Bcl-2表达影响的研究

Expression of Bcl-2 in Human Retinoblastoma Rb Cell Influenced by Allicin

目的探讨大蒜素对人视网膜母细胞瘤Rb细胞的抑制作用,及其对Bcl-2表达的影响。方法人视网膜母细胞瘤Rb细胞株,经复苏,孵育,传代分为10个实验组。分别用不同浓度大蒜素(5μg/ml、10μg/ml、15μg/ml、20μg/ml、25μg/ml、30μg/ml)、顺铂(3μg/ml)及大蒜素(15μg/ml)和顺铂(3μg/ml)联合处理人视网膜母细胞瘤Rb细胞,在不同时间(24h、48h、72h)观察Rb细胞的形态,并应用免疫细胞化学SP法检测不同处理条件下和各时间点Bcl-2的表达情况。结果人视网膜母细胞瘤Rb细胞在不同浓度大蒜素、顺铂以及大蒜素和顺铂联合作用下,细胞生长增殖受到抑制,与大蒜素空白液、培养液对照组的Rb细胞相比形态学变化明显。不同浓度的大蒜素均可以下调人视网膜母细胞瘤Rb细胞中Bcl-2的表达,大蒜素作用组与对照组两两比较,差异均有统计学意义(P<0.05)。大蒜素下调Bcl-2的表达具有时间和剂量依赖性,随着大蒜素作用时间的延长,Bcl-2的表达依次下降。大蒜素(15μg/ml)和顺铂(3μg/ml)联合用药可增强对人视网膜母细胞瘤Rb细胞的生长抑制,与大蒜素(15μg/ml)、顺铂(3μg/ml)单独处理Rb细胞比较,下调Bcl-2的表达作用具有显著性差异(P<0.05)。结论人视网膜母细胞瘤Rb细胞中存在与细胞凋亡有关的Bcl-2蛋白低表达。大蒜素可抑制人视网膜母细胞瘤Rb细胞的生长,且抑制作用随药物作用浓度的增高和作用时间的延长而增强,具有时间和浓度依赖性。采用免疫细胞化学的方法检测人视网膜母细胞瘤Rb细胞中Bcl-2的表达,发现大蒜素作用组Bcl-2的表达较空白和溶剂对照组低,提示大蒜素抑制人视网膜母细胞瘤Rb细胞的生长与Bcl-2介导的细胞凋亡有关,大蒜素和顺铂联合用药作用强于单独用药,可进一步提高药物对肿瘤细胞的生长抑制。

Objective To research the exhibition on retinoblastoma Rb cell of allicin, and the impact of Bcl -2 expression. Methods Human retinoblastoma Rb cell was divided to 10 experiment groups through resuscitation and incubation. Different concentrations of allicin （5 μg/ml, 10μg/ ml, 15 μg/ml,20μg/ml,25 μg/ml,30μg/ml） disposes Human retinoblastoma Rb cell, cisplatin （ 3 μg/ ml） disposes Human retinoblastoma Rb cell, allicin（15μg/m） plus cisplatin（3μg/ml） disposes Human retinoblastoma Rb cell, observing the shapes of Human retinoblastoma Rb cell in different time （24 h, 48h, 72h）, At the same time, adopting immunocytochemistry SP method to detect the expressions of Bcl -2. Results Human retinoblastoma cells in different concentrations of allicin, cisplatin and allicin combined with cisplatin, cell growth and proliferation were inhibited. There were obvious morphological changes compared to the control group. Different concentrations of allicin can reduced the expression of Bcl -2 in human retinoblastoma Rb cell. There were significant difference between allicin group and the control group （P 〈 0. 05 ）. Allicin reduced expression of Bcl - 2 depending on time and dose manner, by the time, the expression of Bcl - 2 were declining. Allicin （ 15 μg/ml） combined with cisplatin （3 μg/ml） can enhance inhibition of the human retinoblastoma Rb cell growth. Comparing with allicin （15μg/ml）, cisplatin （3μg/ml） alone processing, there were significant difference in the role of reduced the expression of Bcl - 2 （ P 〈 0. 05 ）. Conclusion The expressions of Bcl - 2 showed an obvious reducing in human retinoblastoma Rb cell. Allicin can inhibit human retinoblastoma Rb cell growth, and the inhibition depended on the increasing of drug concentration and the extension of time. Using immunocytochemical methods for detection of retinoblastoma Rb cell, the expression of Bcl -2 in allicin group was lower than that in control group, suggesting that inhibition of retinoblastoma Rb cell growth was related to apoptosis mediated by Bcl -2. Allicin combining with cisplatin can be further enhanced the inhibition of tumor cells growth than that separate using.