rhddADAM15抑制肝癌细胞Bel-7402增殖的作用研究  

Effect and Mechanism of rhddADAM15 on the Proliferation of Bel-7402 Cells

作　　者：侯颖[1] 储敏[1] 王晓敏[1] 陈蕴[1] 金坚[1] 

机构地区：[1]江南大学药学院,药物设计与分子药理实验室,无锡214122

出　　处：《中国细胞生物学学报》2014年第4期415-420,共6页Chinese Journal of Cell Biology

摘　　要：ADAM15属于跨膜蛋白ADAM家族中的一员,在乳腺癌、宫颈癌、卵巢癌等多种实体瘤中均发现其表达量提高。ADAM15在降解细胞外基质、介导细胞的黏附、细胞间信号转导及在肿瘤发展进程中起到重要作用。因其去整合素区域含有RGD序列,ADAM15可与多种整合素相互作用。在前期工作中,实验组利用大肠杆菌表达系统表达了重组人ADAM15去整合素区域蛋白,记作rhddADAM15。该研究将进一步针对rhddADAM15抑制肝癌细胞Bel-7402增殖的机理进行分析及探讨。SRB法显示,rhddADAM15可抑制Bel-7402细胞增殖并呈剂量依赖性,IC50为1.14μmol/L;利用DAPI核染发现,rhddADAM15可显著诱导Bel-7402细胞凋亡;流式细胞仪分析发现,rhddADAM15浓度为6μmol/L时,(87.44±7.25)%的细胞发生凋亡;PI单染分析细胞周期表明,rhddADAM15作用后,部分Bel-7402细胞周期被阻滞于S期及G2/M期,并呈剂量依赖性,4μmol/L rhddADAM15处理后G0/G1期含量下降约14%;Western blot分析显示,rhddADAM15可下调Bel-7402细胞周期蛋白CDC2的表达,抑制CDC2-Tyr15的去磷酸化,引起G2/M期的阻滞。ADAM15, a member of transmembrane proteins ADAMs, is over-expressed in many solid tumors such as breast, colorectal and ovarian cancer, playing important roles in the degradation of extracellular matrix, cell adhe- sion, intracellular signal transduction and pathological changes in tumors. Because of the RGD motif in its disintegrin domain, ADAM15 is considered to interact with integrins multifariously. We have established the recombinant human disintegrin domain of ADAM15 （rhddADAM15） by E.coli in our previous research. This study aimed at assessing the ef- fect and mechanism of rhddADAM 15 on the proliferation of Bel-7402 cells, rhddADAM15 inhibited the proliferation of Bel-7402 with an IC50 of 1.14 pmol/L by SRB assay. Bel-7402 cells had apoptotic amorphological nucleus changes using DAPI staining, and （87.44：t：7.25）% of Bel-7402 cells showed apoptotic features when treated with 6 lamol/L rhddAD- AM 15 analyzed by PI staining. Moreover, partial S and G2/M arrests were observed on Bel-7402 cells. When treated with rhddADAM15 at the concentration of 4 pmoFL, the G0/G1 phase of Bel-7402 cells reduced by 14%. The level of CDC2 was down-regulated and the phosphorylation of CDC-Tyr15 was increased which aroused the G2/M arrest.

关 键 词：ADAM15 去整合素 凋亡 CDC2 

分 类 号：R735.7[医药卫生—肿瘤]