【摘要】 探讨C225单用及与DDP联用对人子宫颈癌HeLa细胞的体外抑制作用及其机制。分别用C225(100、300μg/ml)与DDP(0.1、0.5、1μg/ml)以单用和联用处理HeLa细胞72 h后,MTT法检测对HeLa细胞生长的抑制作用及协同效应;FCM技术检测C225(300μg/ml)与不同工作浓度DDP单用及联用72 h后对细胞周期及凋亡的影响;并用HOE-CHEST33258染色技术进行细胞凋亡的形态学检测。结果C225能明显抑制HeLa细胞增殖,与DDP联合具有协同作用或相加作用;C225能使细胞周期阻滞于G1期,并诱导细胞凋亡;与DDP联用后凋亡率更高,使细胞周期进一步阻滞于G2期,同时降低S期细胞比例。荧光显微镜下可见实验对照组所发荧光较弱,无凋亡小体出现,而各药物干预组均出现一定数量的凋亡细胞,并随药物浓度的增加而增多,其中尤以联合用药组最为明显。两药单用及联用均对HeLa细胞的体外生长具有显著抑制作用,C225可通过将细胞周期阻滞于G0-G1期及诱导细胞凋亡发挥其抑瘤效应;两药间存在协同作用,机制可能与C225增加HeLa细胞对DDP的敏感性有关。更多还原

【Abstract】 To explore the mechanism of single use of monoclonal antibody C225 or combined use with DDP,100 μg/ml or 300 μg/ml of monoclonal antibody C225 was singly or in combination with DDP was applied to the cultures of HeLa cells and the inhibitory effect upon the cellular growth was observed 72 hours later.Control without any treatment was also set up.MTT assay was used to detect the inhibitory effect on the growth HeLa cells and the synergistic effect,and flow cytometry technique was applied to find out the influence of 300 μg/ml of C225 singly used or in combined use of various concentrations of DDP upon the cell cycles and apoptosis.Staining technique with HOECHEST33258 was used to detect the morphological changes in cell apopotosis.It was found that monoclonal antibody C225 showed apparently inhibitory effect on the proliferation of HeLa cells and the synergistic or additive effect could be demonstrated in the combined use with DDP.In addition,C225 could also arrest the cell cycle in G1 phase and induce the cell apoptosis,in which the rate of apoptosis was even higher in case of the combined use with DDP and it could further arrest the cell cycle in G2 phase.and reduce the proportion of S-phase.Under fluorescent microscopic examination,the intensity of fluorescence after treatment was weaker than that of the controls,but no apoptotic bodies could be demonstrated.However,certain amount of cells with apoptosis could be found after treatment,and the rate of apoptosis increased with the increased concentration of drug used,in which it appeared to be most prominent in case of the combined use with DDP.From these observation,it suggests that the mechanism of inhibitory effect of monoclonal antibody C225 singly used or in combined use with DDP may be associated to its blocking effect on the cell cycle in G0 to G1 phase and the induction of apoptosis.更多还原