Suppression of tumorigenicity and metastasis of human renal carcinoma cells by infection with retroviral vectors harboring the murine inducible nitric oxide synthase gene
Suppression of tumorigenicity and metastasis of human renal carcinoma cells by infection with retroviral vectors harboring the murine inducible nitric oxide synthase gene
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DOI：
10.1089/hum.1998.9.6-845
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发表时间：
1998-04-10
影响因子：
4.2
通讯作者：
Fidler, IJ
中科院分区：
文献类型：
--
作者：
Juang, SH;Xie, KP;Fidler, IJ
本研究的目的是确定是否逆转录病毒介导的小鼠巨噬细胞诱导型一氧化氮合酶（iNOS）基因转移可以抑制人肾癌细胞的致瘤性和转移。在pLXSN逆转录病毒载体中构建编码鼠巨噬细胞iNOS的逆转录病毒载体，其中iNOS基因在长末端重复启动子的控制下，新霉素抗性基因在内部猿猴病毒40启动子的控制下。用对照或iNOS逆转录病毒感染高转移性人肾癌SN 12 PM 6细胞，通过北方和Western印迹分析证实iNOS的表达，并且功能性iNOS蛋白，即，一氧化氮（NO）的产生通过测量培养物上清液中亚硝酸盐的积累来确定。未感染或对照细胞在裸鼠的肾脏中产生大的原位肿瘤和大量的实验肺转移，而iNOS感染的细胞在肾脏中产生小肿瘤和很少或没有肺转移，这些数据表明，携带鼠iNOS基因的逆转录病毒感染人肾癌细胞可诱导高水平NO的产生，这与自身细胞毒性、致瘤性抑制、和消除转移。
The purpose of this study was to determine whether retrovirus-mediated transfer of the murine macrophage inducible nitric oxide synthase (iNOS) gene can inhibit tumorigenicity and metastasis of human renal cancer cells. Retroviral vectors encoding murine macrophage iNOS were constructed in the pLXSN retroviral vector with the iNOS gene under the control of a long terminal repeat promoter and a neomycin resistance gene under the control of an internal simian virus 40 promoter. Highly metastatic human renal carcinoma SN12PM6 cells were infected with control or iNOS retrovirus, Expression of iNOS was confirmed by Northern and Western blot analyses, and expression of the functional iNOS protein, i.e., production of nitric oxide (NO), was determined by measuring nitrite accumulation in culture supernatants, Noninfected or control cells produced large orthotopic tumors in the kidney of nude mice and a larger number of experimental lung metastases, whereas iNOS-infected cells produced small tumors in the kidneys and few to no lung metastases, The data indicate that the infection of human renal cancer cells by retroviruses harboring the murine iNOS gene can induce the production of high levels of NO, which is associated with autocytotoxicity, suppression of tumorigenicity, and abrogation of metastasis.