摘要
用重组PCR技术得到Kininogen D5和TRAIL的融合编码序列,将该DNA片段克隆到原核表达载体pMAL-c2,重组质粒转化大肠杆菌BL21,IPTG诱导蛋白表达,得到MBP-KT和MBP-TK,经AmyloseResin亲和层析柱层析,得到初步纯化的融合蛋白。结果表明MBP-KT对胰腺癌细胞1990有明显的杀伤作用其作用的ED50为20 ng/ml,而MBP-TK对1990的抑制作用不明显;同时,MBP-KT和MBP-KD5对内皮细胞ECV304有明显的抑制作用,MBP/KT作用于ECV304的ED50为0.1μg/ml,MBP/KD5作用于ECV304的ED50为9.5μg/ml,但是MBP-TK和TRAIL几乎无作用,表明融合蛋白KT既具抗肿瘤作用又有抗新生血管的作用。本研究为进一步开发靶向性杀伤肿瘤药物奠定了基础。
The fused gene of Kininogen D560148 and TNF-related apoptosis-inducing ligand(TRAIL114-281) were amplified by PCR, and was inserted into pMAL-c2 vector. The recombinant plasmid pMAL-KT and pMAL-TK were transformed into E. coli BL21. After IPTG induction, the recombinant MBP-KT and MBP-TK proteins in partially purified forms were obtained respectively. The recombinant products were tested for their cytotoxic or cytostatic activities against SW1990 and ECV304 cells. The results showed that MBP-KT plays a significant cytotoxicity against SW-1990 cells, but MBP-TK had no evident effect. Meanwhile, it exhibited also a growth inhibitory effect on proliferation of ECV304 cells. The results suggested that fusion protein MBP-KT had an antiproliferative effect on cancer cells as well as an inhibition effect on angiogenesis. This research provided the foundations of the target therapy in cancer treatment.
出处
《药物生物技术》
CAS
CSCD
2008年第1期1-5,共5页
Pharmaceutical Biotechnology
基金
上海市现代生物与新药产业发展基金(024319115)