蛋白酶体抑制剂Velcade诱导肝癌细胞HepG2凋亡  

Velcade induced apoptosis in HepG2 cells

作　　者：陈锡林[1] 汪谦[1] 江穗[2] 曹良启[3] 黄晓卉[1] 谭浩翔[1] 陈劲松[1] 

机构地区：[1]中山大学附属第一医院普通外科,广州510080 [2]广东省人民医院 [3]广州医学院附属第二医院肝胆外科,广州510260

出　　处：《中华普通外科学文献（电子版）》2008年第5期17-19,共3页Chinese Archives of General Surgery(Electronic Edition)

基　　金：广东省科技计划项目(2007B031514005)

摘　　要：目的探讨蛋白酶体抑制剂Velcade诱导人肝癌细胞株HepG2凋亡的作用和机制。方法以不同浓度Velcade处理HepG2细胞24h和48h。四甲基偶氮唑蓝比色法评价细胞生长情况;流式细胞术检测细胞凋亡;Western blot测定caspase 3改变;RT-PCR检测Bcl-2家族、Cyclin A和Cyclin D mRNA的表达。结果不同Velcade浓度(32、64、128、256、512nmol/L)处理HepG2细胞48h后,细胞活性较对照组明显减少(分别是91.4%±2.1%、75.0%±3.7%、57.3%±1.6%、52.7%±1.6%和31.4%±2.6%),与对照组(100.0%±1.7%)比较具有显著性差异(P<0.05)。128nmol/L的Velcade处理HepG2细胞,48h流式细胞仪检测结果显示SubG1细胞百分数明显升高,SubG1细胞百分数(27.3%±5.3%),与对照组(4.3%±0.5%)差别显著(P<0.05),同时,Pro-caspase 3明显下降;但Velcade对Bcl-2家族表达无明显影响;Velcade诱导HepG2细胞G2/M阻滞,抑制Cyclin A和Cyclin D表达。结论Velcade诱导肝癌细胞凋亡不通过改变Bcl-2家族表达,可能存在其它途径;Velcade诱导肝癌细胞G2/M周期阻滞与调节通过下调Cyclin A和Cyclin D表达相关。Objective To investigate the effects of Velcade, a proteasome inhibitor, on HepG2 cells, and to explore the mechanism. Methods HepG2 cells were treated with various concentrations of Velcade for 24 h and 48 h. The cell viability was assessed by 3-（4,5-Dimethyhhiazol-2-yl）-2,5-diphenyl tetrazolium bromide （MTT assay. Cell.apoptosis was investigated by flow cytometry. Western hlot assayed the change of easpase 3. The expressions of Bcl-2 family, Cyclin A and Cyclin D were detected by reverse transcription-polymerase chain reaction（RT-PCR）. Results Viabilities of HepG2 cells treated with various concentrations of Velcade （32, 64, 128, 256, 512 nmol/L） for 48 h were significantly decreased compared to control group. Viabilities of HepG2 cells gradually reduced to 91.4% ± 2.1%, 75.0%± 3.7%, 57.3% ±1.6%, 52.7% ± 1.6% and 31.4% ± 9.6%, respectively. The differences compared with＇control group （100.0% ± 1.7%） were significant （P〈0.05）. The percent of SubG1 of HepG2 ceils （27.3% ± 5.3%） treated with 128 nmol/L Velcade 48 h was significantly higher than that in control group （4.3% ± 0.5%, P〈0.05）. After treatment with Velcade （128 nmol/L） for 24 h, the expression of caspase 3 markedly decreased compared with control. Velcade greatly increased the G2/M phase in HepG2 cells. Velcade resulted in no significant changes of Bcl-2 family member. However, Velcade down regulated the expressions of Cyclin A and Cyclin D. Conclusion The mechanism of Velcade to induced apoptosis in HepG2 is not associated with change of Bcl-2 family member. Velcade induced G2/M phase arrest via down regulation of Cyclin A and D.

关 键 词：蛋白酶体抑制剂 VELCADE 肝癌 凋亡 

分 类 号：R735.7[医药卫生—肿瘤] R979.1[医药卫生—临床医学]