目的：通过构建人血管能抑制素（Canstatin）真核表达载体，转染裸鼠肿瘤局部，探索其对肺癌治疗作用。方法: RT-PCR法获取Canstatin cDNA全长，定向克隆法构建Canstatin基因表达载体pCMV-Script-Cans。荧光定量PCR法检测转染细胞Canstatin mRNA的表达。台盼蓝拒染法、3H-TdR掺入法检测细胞生长增殖，TUNEL法检测细胞凋亡。基因枪转染重组载体到荷瘤裸鼠肿瘤局部，CD31单克隆抗体微血管记数检测抗血管生成效应。结果：成功构建pCMV-Script-Cans重组载体，并在转染的细胞株中检测到Canstatin mRNA的表达。人脐静脉内皮HUV-EC-C细胞株pCMV-Script-Cans质粒转染组比空载体组3H-TdR掺入量明显减低（P<0.01），细胞凋亡率明显增加（P<001），重组载体转染肿瘤生长缓慢，微血管数显著低于对照组（P<0.01）。结论pCMV-Script-Cans重组载体能在转染的哺乳动物细胞中表达Canstatin，并抑制内皮细胞增殖，而且有很好的抗肺癌血管生成作用。

Objective:To construct a mammal expression system of human canstatin and study its anti-tumor effects on lung cancer. Methods:Canstatin cDNA was acquired by RT-PCR, and cloned into a mammal exprssion vector named pCMV-Script. Canstatin expression was detected by Real-time PCR. The proliferation, apotosis of the cells transfected with recombinant canstatin vector were measured by trypan blue exclusive assay, 3H-thymidine incorporation and TUNEL method respectively. Then the recombinant vector encoding canstatin cDNAs was transferred into tumors of cancer-bearing nude mice with electroporator in vivo, and micro-vessel count was proceeded of each tumor by anti-CD31 antibody immunohistochemical staining.Results: The recombinant vector pCMV-Script-Cans was successfully constructed , and the canstatin mRNA was detected in both of the transformed HUVE and A549 cells. The 3H-TdR intake rate in pCMV-Script-Cans transformed HUVE cells is significant lower than that of the naked plasmid transformed cells (P<0.001) , while the apotosis rate of them is significant higher than that of the control cells(P<0.001). The micro-vessels in the recombinant vector transformed tumors were significant lower than that of the control group. Conclusions: Canstatin only inhibit cell proliferation and induce apotosis in endothelial cell, and it also has a good anti-tumor effect in vivo.