诱导型一氧化氮合酶基因转染对人舌癌细胞体内致瘤力的影响

Effects of inducible nitric oxide synthase on tumorigenesis and biological characteristics in human oral cancer cell

目的研究外源性基因诱导型一氧化氮合酶(iNOS)转染人舌癌细胞系Tca8113后,对肿瘤细胞生物学特性以及裸鼠体内致瘤力的影响。方法采用脂质体转染法将真核表达载体pCMV-iNOS/Tca转染入舌癌细胞系Tca8113,得到高表达iNOS的细胞系pCMV-iNOS/Tca,转染空白质粒的细胞系pCMV-Tca,并以Western blot方法鉴定转染效果。四甲基偶氮唑蓝(MTT)比色绘制生长曲线;流式细胞仪检测细胞周期分布;通过裸鼠成瘤实验,比较转染前后体内成瘤能力的差别。结果与未转染组和转染空白载体组比较,pCMV-iNOS/Tca组的细胞生长速度比Tca组快(P>0.05),细胞周期中G2/M和S期比例增加(P<0.05)。体内成瘤能力试验结果显示,pCMV-Tca、pCMV-iNOS/Tca组和Tca组平均瘤重分别为(1.1±0.24)g、(2.5±0.48)g和(1.3±0.32)g,pCMV-iNOS/Tca组的成瘤力高于pCMV-Tca和Tca组(P<0.01)。结论转染iNOS基因后,Tca8113细胞具有更强的体外增殖能力和体内致瘤能力。

Objective To investigate the effects of inducible nitric oxide synthase （iNOS） on tumorigenesis and biological characteristics in human oral cancer cell line Tca8113. Methods Tca8113 cell was transfected with murine pCMV-iNOS/Tca by lipofectin. Stable transfecrants that highly express iNOS were selected and named pCMV-iNOS/Tca, and pCMV - Tca was defined as cells transfected with empty vector. The transfection efficiency was detected by Western blot. Cell growth curve and cell cycle were measured by MTT assays and flow cytometry. The effects of iNOS on tumor growth were evaluated in vivo by using Tca8113 animal model. Results iNOS transfected cells grow quickly than those of the vector only transfectant and wild type. Moreover, increased G2/M and S phase cells were observed in iNOS transfectant in comparison with the other two groups. The mean tumor weight in iNOS group was（ 2.5 ± 0.48） g, which was significant higher than that of vector only group（ 1.1 ± 0.24） g and wild group （ 1.3 ± 0.32） g. Discussion iNOS may promote cell proliferation in vitro and increase tumorgenesis in vivo in Tca8113 human oral cancer cell line.