摘要
将编码人 94个氨基酸的前列腺分泌蛋白 ( PSP94) c DNA与酵母整合载体 p PICZαA重组 ,构建的重组质粒线性化后转染酵母细胞 GS1 1 5,获得了 PSP94在酵母细胞中遗传性稳定表达酵母工程细胞 .诱导后的培养物中 ,rh PSP94表达量约为 0 .9mg/L,分子量约 1 6.5k D.培养上清经离子交换层析纯化后 ,目的蛋白的纯度为 92 % .体外在人前列腺癌细胞上活性分析表明 ,rh PSP94以1 0 0μg/L ,对该细胞的抑制率 2 0 .4% ;单纯新型 TNF,以 1 0 3 U/ml,抑制率 2 9.8% ;rh PSP94和新型 TNF以上述同样剂量联合应用 ,抑制率为 86.3% .提示 PSP94在体外对抗前列腺癌细胞有杀伤作用 ,但不明显 ;PSP94与新型 TNF联合应用 ,可使抑制率明显提高 ,可能 PSP94与新型 TNF有协同抗前列腺癌的作用 .
cDNA encoding human prostate secretory protein(PSP) of 94 amino acids was inserted into yeast integrative vector pPICZαA and yeast cell GS115 was transfected by linear recombinant plasmid,then the yeast engineering cells were obtained stably expressing PSP94.Expression amount of rhPSP94 after induction was 0.9 mg/L,and molecular weight 16.5 kD.The purity of the title protein got from the medium supernatant after purified by ion exchange chromatography was 92%.The activity analysis of PSP 94 on human prostate carcinoma cells in vitro shows that inhibitory rate was 20.4% when rhPSP94 (100 μg/L) was applied;inhibitory rate was 29.8% when single TNF derivative of 10 3 U/ml was applied;inhibitory rate was 86.3% when combination of PSP 94 and TNF derivative was applied.The results indicate that PSP94 has the killer effect on human prostate carcinoma cells in vitro ,but not remarkable;combination of PSP94 and TNF derivative can increase the inhibitory rate.The possible reason is that PSP94 and TNF derivative have the cooperative effect on prostate carcinoma.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
2000年第4期542-545,共4页
Chinese Journal of Biochemistry and Molecular Biology
关键词
前列腺分泌蛋白
TNF衍生物
前列腺癌
抗肿瘤
Prostate secretory protein of 94 amino acids, TNF derivative, Prostate carcinoma, Antitumor
