HSV_1-TK基因转导人肺腺癌细胞A549体内外表达的研究

Studies on the expression of HSV1TK gene in vitro and in vivo after transduction into human pulmonary carcinoma cell line A549

目的：观察ＴＫ基因转导Ａ５４９细胞后在体外和体内的表达。方法：将已构建的逆转录病毒表达载体ＰＬＸＳＮ－ＴＫ用电穿孔法转化Ａ５４９细胞，原位杂交检测ＴＫｍＲＮＡ在Ａ５４９－ＴＫ细胞和由它接种裸鼠形成肿瘤组织中的表达。斑点杂交检测外源基因在细胞中整合。并体外观察Ａ５４９－ＴＫ细胞对ＧＣＶ的敏感性。结果：原位杂交表明Ａ５４９－ＴＫ细胞及由其所形成的肿瘤组织中ＴＫｍＲＮＡ表达阳性，对照细胞无表达，斑点杂交证明Ａ５４９－ＴＫ细胞中有ＴＫ基因整合。Ａ５４９－ＴＫ细胞对ＧＣＶ的敏感性是亲代细胞的４６倍。结论：ＴＫ基因在Ａ５４９－ＴＫ细胞中表达阳性，转基因细胞对ＧＣＶ敏感。

Objective: To observe the expression of TK gene in vitro and in vivo after it being transduced into A549 cells. Methods: The constructed retroviral vector PLXSNTK was transduced into A549 cells by electrotransformation. The TK mRNA expression in A549TK cells and tumor tissue formed by itself in nude mice was determined with in situ hybridization. Recombination of TK gene into cells is examined with blot hybridization. Sensitivity of A549TK cells to GCV was also observed in vitro. Results: In situ hybridization showed that A549TK cells and tumor tissue positively expressed TK mRNA while the control cells did not. Blot hybridization showed that TK gene was recombined in A549TK cells. The sensitivity of A549TK cells to GCV was 46 times higher than that of parental cells. Conclusion: TK gene is positively expressed in A549TK cells and the sensitivity to GCV is also obtained by A549TK cells.