组蛋白去乙酰化酶抑制剂SAHA联合溶瘤腺病毒ZD55-IL-24诱导SW480细胞凋亡  

Apoptosis in SW480 Cells Induced by Combining Oncolytic Adenovirus ZD55-IL-24 with Histone Deacetylase Inhibitor SAHA

作　　者：谌贺宽子[1] 梁天祥[1] 仇庆[1] 夏玉龙[1] 吴欣欣[1] 周秀梅[1] 

机构地区：[1]浙江理工大学生命科学学院新元医学与生物技术研究所,杭州310018

出　　处：《中国细胞生物学学报》2012年第1期54-60,共7页Chinese Journal of Cell Biology

基　　金：浙江理工大学科研启动基金(No.1016834-Y)资助项目~~

摘　　要：研究组蛋白去乙酰化酶抑制剂SAHA联合溶瘤腺病毒ZD55-IL-24对结肠癌SW480细胞的体外杀伤作用。采用MTT法、结晶紫实验检测SAHA、ZD55-IL-24以及二者联合使用对结肠癌细胞株SW480及人正常肺上皮细胞株Beas-2B的增殖抑制作用;利用Hoechst33342染色对经各种处理的细胞进行凋亡形态学观察,采用流式细胞术对凋亡进行量化;通过Western blot法在蛋白水平上检测SW480细胞中IL-24的表达情况。结果显示SAHA与ZD55-IL-24联合处理对SW480的增殖抑制作用明显优于两者单独使用。10 MOI病毒ZD55-IL-24与0.5μmol/L SAHA联合作用4天,SW480细胞存活率仅为12%,明显低于10 MOI病毒单独处理组细胞的存活率(40%,P<0.05)。然而,正常细胞对于联合给药显示出良好的耐受性。Hoechst33342染色和流式细胞术结果也表明联合处理组的SW480细胞凋亡特征更明显。此外,IL-24在ZD55-IL-24病毒单独感染组及病毒与药物联合组的SW480细胞中均能有效表达。To investigate the anti-tumor effects in colorectal cancer cell line SW480 in vitro by combining oncolytic adenovirus ZD55-IL-24 with suberoylanilide hydroxamic acid(SAHA),MTT assay and crystal violet assay were used to determine the growth inhibition effect of single or combination therapy on colorectal cancer cell line SW480 and normal human lung epithelial cell line Beas-2B.After Hoechst33342 staining,the morphological changes of apoptosis in treated cells were observed under fluorescence microscope.And flow cytometry assay was used to quantify apoptosis.The expression level of IL-24 protein was assayed by Western blot.The results of MTT and crystal violet staining showed that the growth inhibition of SW480 cells treated with the combination of ZD55IL-24 and SAHA were more obvious than that in ZD55-IL-24 group or SAHA group alone.Four days after administration of 10 MOI ZD55-IL-24 and 0.5 μmol/L SAHA,the cell viability rate of SW480 cells was only 12%,which was significantly lower than that of cells treated only by 10 MOI ZD55-IL-24(40%,P<0.05).In contrast,Beas-2B cells were spared from inhibition of combined treatment(P<0.05).Hoechst33342 staining and flow cytometry also confirmed that there were much more apoptotic SW480 cells in combinational treatment group.In addition,IL-24 protein was effectively expressed in SW480 cells infected with ZD55-IL-24 alone or treated with ZD55-IL-24 plus SAHA.

关 键 词：SAHA 溶瘤腺病毒 IL-24 结肠癌 流式细胞术 细胞凋亡 

分 类 号：R735.35[医药卫生—肿瘤]