HSV-tk/GCV系统对高转移性粘液表皮样癌细胞系M3SP2的抑制作用  

Inhibitory effects of the herpes simplex virus-thymidine kinase gene/ganciclovir system on the highly metastatic mucoepidermoid carcinoma cell line M3SP2

作　　者：刘斌[1] 吴军正[1] 司徒镇强[1] 王江华[1] 刘杰忠[1] 

机构地区：[1]第四军医大学口腔医学院口腔生物学教研室,陕西西安710033

出　　处：《第四军医大学学报》2003年第12期1078-1081,共4页Journal of the Fourth Military Medical University

基　　金：第四军医大学科技创新工程课题 (CX99A0 1 1 )

摘　　要：目的 :探讨单纯疱疹病毒胸苷激酶基因 (HSV tk) /丙氧鸟苷 (GCV)自杀基因系统对高转移性粘液表皮样癌细胞系M3SP2的作用 .方法 :采用阳离子脂质体介导法将含HSV tk基因复制缺陷型逆转录病毒载体 (pBabe/tk)导入M3SP2细胞 ,嘌呤霉素筛选抗性克隆 .用MTT比色法测定HSV tk/GCV对tk阳性M3SP2细胞 (M 3SP2 /tk)生长的影响及旁观者效应 ;用流式细胞术分析HSV -tk/GCV对M3SP2 /tk细胞周期的影响 .结果 :M3SP2 /tk细胞对GCV的敏感性显著增加 ,细胞生长受到抑制 ,在GCV(0 .1～ 10 0 0mg·L-1)作用 6d时 ,M3SP2 /tk细胞存活率为 6 2 .3%- 14 .5 %,IC50 值为 0 .6 8mg·L-1,而转空载体细胞和亲本细胞IC50 值大于 10 0 0mg·L-1.M3SP2 /tk细胞在GCV(1～ 10mg·L-1)作用 4 8h时 ,明显阻滞于S期 .10 %tk阳性的M 3SP2 /tk细胞与 90 %tk阴性的亲本细胞M3SP2混合培养 ,10mg·L-1的GCV仍可杀伤 5 4 .4 %的细胞 .M3SP2 /tk细胞含GCV (1,10mg·L-1)培养上清液仍能影响tk阴性的舌癌细胞系TbpTL的生长 .结论 :HSV tk/GCV具有显著抑制涎腺粘液表皮样癌细胞体外生长的作用 。AIM: To explore inhibitory effect of the herpes simplex virus thymidine kinase gene (HSV tk ) plus ganciclovir (GCV) system on the highly metastatic mucoepidermoid carcinoma cell line M3SP2 from human salivary gland. METHODS: The M3SP2 cells were transducted by the replicative deficient retroviral vector containing the HSV tk gene with the cationic liposomes. The cytotoxicity and the bystander effect of the HSV tk /GCV system on the M3SP2 cells were examined by MTT assay. The effect of HSV tk /GCV on cell cycle of the M3SP2 cells transducted with HSV tk gene (M3SP2/ tk ) was tested by the flow cytometry. RESULTS: The sensitivity of the M3SP2/ tk cells to GCV was increased obviously. When the M3SP2/ tk cells were exposed to GCV at the concentration of 0.1-1000 mg·L -1 for 6 days, the cell survival rate of M3SP2/ tk cells were 62.3% -14.5% and the 50% of the inhibitory concentration (IC 50 ) was 0.68 mg ·L -1 , whereas the IC 50 of the parent cells or of the cells transducted with blank vector was more than 1000 mg·L -1 . The M3SP2/ tk cells treated with GCV at the dose of 1-10 mg·L -1 for 48 h could be significantly arrested in S phase. Moreover, even when only 10% of the tk + M3SP2/ tk cells plus 90% of tk tongue cancer cells TbpTL, 54.4% of the mixed cells could be killed while exposed to GCV (10 mg ·L -1 ) and the supernatant from M3SP2/ tk cells treated with GCV (10 mg ·L -1 ) could induce growth inhibition of tk tongue cancer cell line TbpTL. CONCLUSION: The HSV tk /GCV system can inhibit in vitro growth of the mucoepidermoid carcinoma cells, and its mechanism of cell killing is related to the arrest of cell cycle progresss and the bystander effect.

关 键 词：癌 粘液表皮样 涎腺 单纯病毒属 胸苷激酶 旁观者效应 基因疗法 

分 类 号：R736[医药卫生—肿瘤]