摘要
观察重组人ADAM15去整合素区域蛋白干预肿瘤细胞增殖效应,为研究其作用机制提供基础。采用SRB法筛选重组人ADAM15去整合素区域蛋白敏感肿瘤细胞,分析其剂量依赖关系。利用流式细胞仪分析重组人ADAM15对肿瘤细胞周期的影响,并利用DAPI染色检测其对肿瘤细胞凋亡的诱导作用。重组人ADAM15去整合素区域蛋白对于观察的8种肿瘤细胞的增殖均有明显的抑制作用,其中人宫颈癌细胞Hela及野生型乳腺癌细胞MCF-7最为敏感,IC50分别为2.97,2.90μmol/L,当其浓度低于8μmol/L时,细胞增殖抑制率与蛋白浓度呈剂量依赖关系。选择Hela细胞进行周期检测发现,重组人ADAM15去整合素区域蛋白浓度高于2μmol/L时,Hela细胞周期开始出现S期阻滞,G2/M期含量下降,并出现核凋亡现象。当蛋白干预浓度达6μmol/L时,(53.57±1.43)%的Hela细胞出现凋亡。重组人ADAM15去整合素区域蛋白通过阻滞细胞分裂和诱导凋亡抑制肿瘤细胞的增殖。
ADAM15, a transmembrane ADAM, which is usually expressed on smooth-muscle, mesangial and endothelial cells, is over-expressed in many solid tumors such as breast, colorectal, and ovarian cancer . It plays important roles in the degradation of extracellular matrix, cell adhesion,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. The recombinant human disintegrin domain of ADAM15 （rhddADAM15 had been expressed） by E. coli in our previous research . This study aimed at assessing the effect of recombinant disintegrin domain of ADAM15 （ rhddADAM15 ） on the proliferation of different tumor cells and selecting the sensitive cells for the further study. The SRB assay was used to determine the effect of rhddADAM15 on the proliferation of different tumor cells and the DAPI staining was used to detect the morphological changes of the nuclei. The cell cycle was analyzed using flow cytometer. It was found that the rhddADAM15 had strong inhibitory effect on the proliferation of the 8 tested tumor cells and the most sensitive cells were human cervical carcinoma cell Hela and human breast cancer cell MCF-7, with ICs0 of 2.97 and 2.90 μmol/L respectively. When the concentration of rhddADAM15 was lower than 8 μmol/L, the inhibitory rate was in a dose-dependent manner. The Hela cell was chosen and the cell cycle and nuclear morphology were evaluated. After treated with rhddADAM15, the cells had a partial S phase arrest analyzed using flow cytometer. In addition ,the nuclei of Hela cells had morphological changes of apoptosis and the number of apoptotic cells increased in a dose-dependent manner. In conclusion, rhddADAM15 inhibited the proliferation of different tumor cells and induced morphological nucleus changes of apoptosis and a partial S phase arrest of Hela cells.
出处
《药物生物技术》
CAS
2014年第2期95-98,共4页
Pharmaceutical Biotechnology
