Cloning Of Human Endostatin Gene And Expression In Hela Cell

Endostatin, a carboxy-terminal 20KDa fragment of the non-coliagenous (NC) 1 domain of collagen â…©â…§, is the first endogenous angiogenesis inhibitor entering human clinical trials. It can inhabit endothelial cell migration and multiplication to reduce the vascularization of tumors and finally kill tumors. The great advantage of the agent lies in the lack of toxicity and drug resistance. Human Endostatin gene cloned by RT-PCR was inserted into separate expression vector pIRES2-EGFP and fusion expression vector pEGFP-Ci, which were then transfected into hela cell with lipofectamine? regent. After 48 hours, under the fluorescence microscope, the green fluorescence was spreading in the transfected hela cell. After culturing ten generations, the green fluorescence expressed persistently. This illustrated that the transfection was steady. Further more, after immunocytochemistry in the fixed tranfected hela cell with rabbit anti-Endostatin, under the microscope, the transfected hela cell became brown-red and the negative control was colourless, which meant that Endostatin has steadily expressed in the transfected hela cell. Our results provided a basal evidence for gene therapy and gene medicine.