国家重点基础研究发展规划项目(973)(39700158)和国家自然科学基金资助项目(39800142,30100105).
This work was supported by grants from the Special Funds for Major State Basic Research of China (G199980510008) and The National Natural Science Foundation of China (39800142, 30100105).
为了探讨鼻咽癌表达下调基因NAG7对鼻咽癌细胞系HNE1生长的影响, 构建了NAG7基因的真核表达载体pcDNA3.1(+)/NAG7, 并采用脂质体转染技术将真核重组体pcDNA3.1(+)/NAG7质粒和真核空载体pcDNA3.1(+)质粒分别导入HNE1细胞, 经G418筛选后获得稳定转染细胞克隆, RT-PCR和RNA印迹检测NAG7基因的表达, 并通过细胞生长曲线、裸鼠接种和流式细胞等方法对转染细胞的生物学行为进行检测.结果显示:转染NAG7基因后,基因表达增加,细胞生长倍增时间较空载体转染和HNE1明显延长,流式细胞技术检测表明,NAG7可延缓细胞由G0~G1期进入S期;裸鼠接种实验显示转染NAG7基因后的HNE1细胞致瘤性受到抑制.上述结果表明:NAG7基因转染后鼻咽癌细胞生长受到抑制,提示NAG7基因是一鼻咽癌相关的抑瘤基因候选者.
In order to study the effect of NAG7 gene on NAG7 down-regulated nasopharyngeal carcinoma (NPC) cell line HNE1, the pcDNA3.1(+)/NAG7 mammalian expression recombination was constructed and transfected into HNE1 cells. G418 was used to obtain the neomycin-resistant transformants of which indicated the vector was present in the HNE1 cells. The expression of NAG7 gene was detected by RT-PCR and Northern blot. The cytobiologic characteriztions of the transfected HNE1 cells were probed by population double time (PDT), xenograft of nude mice and cell cycle analysis. The results showed that the PDT of G418-resistant HNE1 cells with expression of NAG7 was longer than that of vector transfected HNE1 cells and untranstected HNE1 cells, and the more NAG7 transfected cells went into phase G0-G1 compared with the two other cells. It also presented inhibited the tumor formation in mude mice. Thus, these data suggested that NAG7 gene play an important role in the cell growth of HNE1, and might be a good candidate of tumor suppressor gene correlated with NPC.
谭琛,李江,王洁如,彭聪,谢奕,曹利,李小玲,李桂源. NAG7基因转染对鼻咽癌细胞生长的影响[J].生物化学与生物物理进展,2002,29(3):372-377
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