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NGX6基因是我室利用定位候选克隆策略,在鼻咽癌的高频杂合性缺失区9p21-22克隆的新基因.前期研究结果提示,它与鼻咽癌细胞的侵袭转移密切相关.为了进一步阐明其作用的结构基础,本研究成功构建了含NGX6基因及突变体的pCMV-myc瞬时和pcDNA3.1-his-myc(-)B稳定表达载体.通过脂质体转染技术,构建了NGX6及突变体的稳定表达细胞系5-8F.用免疫荧光试验和免疫电镜观察了NGX6在5-8F细胞中的亚细胞定位主要位于胞膜、核膜以及胞浆中的膜质结构,缺失突变的功能域对其定位没有明显的影响.基质胶侵袭试验和划痕试验研究了NGX6及突变体对细胞运动的影响.NGX6能抑制高转移潜能的鼻咽癌细胞5-8F的运动和侵袭能力,缺失胞内区(CYTO)后NGX6不能抑制5-8F细胞的运动和侵袭,提示CYTO可能是其发挥作用的重要功能域.
NGX6 gene is a new gene cloned by 9p21-22 in high frequency heterozygous deletion region of nasopharyngeal carcinoma, which is a candidate gene cloning strategy in our laboratory.Previous studies suggest that it is closely related to the invasion and metastasis of nasopharyngeal carcinoma cells.In order to further To clarify the structural basis of its role, this study successfully constructed transient expression vector pCMV-myc and pcDNA3.1-his-myc (-) B containing NGX6 gene and its mutants, and constructed the NGX6 And mutants of stable expression cell lines 5-8F.Using immunofluorescence assay and immunoelectron microscopy, the subcellular localization of NGX6 in 5-8F cells was mainly located in the membranous membrane, nuclear membrane and cytoplasm, the deletion mutation Had no significant effect on its localization.Matrix matrigel invasion assay and scratch test investigated the effect of NGX6 and its mutants on cell motility.NGX6 can inhibit the motility and invasion ability of high metastatic potential nasopharyngeal carcinoma cells 5-8F , NGX6 can not inhibit the motility and invasion of 5-8F cells after CYTO is deleted, suggesting that CYTO may be an important functional domain of CYTO.