携带Kallistatin的溶瘤腺病毒载体构建及其对肝癌细胞的体外杀伤效应

Construction of Oncolytic Adenovirus Vector Carrying Kallistatin and Its Killing-Effect to Liver Cancer Cells in Vitro

采用靶向基因-病毒治疗策略,通过同源重组的方式构建了携带Kallistatin(KAL)的重组溶瘤腺病毒ZD55-KAL,并研究其对肝癌细胞的杀伤作用。通过PCR方法鉴定病毒构建正确;MTT法检测病毒对肝癌细胞的杀伤能力和对正常细胞的安全性;结晶紫染色观察细胞凋亡现象。结果显示:经目的病毒ZD55-KAL感染后肝癌细胞出现明显的病变和生长抑制,而正常细胞未出现病变现象,表明目的病毒具有较高的安全性和对肿瘤细胞的特异性杀伤能力。此外ZD55-KAL感染肝癌细胞后引起凋亡,所携带的治疗基因KAL能通过促进肿瘤的凋亡达到抑制肿瘤细胞生长的效果。

Here the authors use targeting gene-virotherapy strategy to construct recombinant oncolytic adenovirus ZD55-KAL harboring Kallistatin（KAL） gene by homologous recombination method, and inves- tigate its antitumor effect to liver cancer ceils. Subsequently, PCR for identification of recombinant virus, MTT assay and crystal violet staining for detection of tumor killing-effect and safe to normal cells is per- formed. The results show that the hepatoma cells have apparent lesions and growth inhibition effect after infection by ZD55 KAL, while no impact on the normal cells showing its high security. In addition, infection by the therapeutic viruses causes apoptosis of hepatoma cells by carrying the KAL gene, indicating the perfect security ability and the effect of inhibition of tumor cell growth.