RAB5A基因表达改变对人肺腺癌细胞系GLC-82和SPC-a1的分化和侵袭特性影响

Studies of the Cellular Biological Function of Expression Change of RAB5A Gene in Human Lung Adenocarcinoma GLC-82 and SPC-al

为探讨RAB5A基因对两种人肺腺癌细胞系GLC 82和SPC al分化及侵袭特性的影响 ,利用细胞转染技术将构建的RAB5A反义RNA重组质粒 (pcDNA3 AntiRAB5A)和RAB5A正义真核表达载体分别转染入低分化人肺腺癌细胞系GLC 82和低转移人肺腺癌细胞系SPC al中 ,在稳定筛选后 ,通过裸鼠体内实验和体外人工基底膜侵袭和细胞趋化运动实验 ,观察转染后细胞分化和转移特性的改变。观察转染前后细胞 ,发现转染后GLC 82细胞体外侵袭重组基底膜能力及趋化运动能力降低 (t检验P <0 .0 2 ) ;裸鼠体内成瘤实验 ,瘤块切片病理观察转染后GLC 82细胞出现腺腔样及基底模样结构 ,分化程度增高。转染后SPC al细胞体外趋化运动能力、侵袭重组基底膜能力均增强 (t检验P <0 .0 2 )。RAB5A基因通过影响细胞的体外趋化运动能力、侵袭重组基底膜能力等对GLC 82和SPC al细胞的侵袭转移表型形成及GLC 82细胞的分化性发挥重要作用。

To determine the effect of RAB5A gene over expression on invasion and differentiation of Human Lung Adnocarcinoma Cells SPC al and GLC 82. Using constructed antisense RNA of RAB5A (pcDNA3-AntiRAB5A) and RAB5A eukaryotic expression vector (pcDNA3.1-RAB5A),we stably transfected them into low differentiation human lung adenocarcinoma GLC 82 cell and human lung adenocarcinoma cells SPC al with low metastasis potential capability respectively in vitro . We observed the change of capability of transfected cells in invading recombinant basic membrane and chemotactic motion experiment in vitro , and we make use of hypodermic method with tumor cells in nude to observe the change of differentiation in transfected GLC 82 cells. The transfected GLC 82 cells with pcDNA3-antiRAB5A showed notable alteration. The capability of invading recombinant basic membrane and chemotatic motion decreased in transfected GLC 82 cells. The differentiation of transfected GLC 82 cells was apparently improved. The array of adenocytes is regular and it appears adenoid structure. After RAB5A eukaryotic expression plasmid was transfected into SPC al ,the invasive activity of cells is increased. Over expression of RAB5A played an important role in invasion and differentiation of human lung adenocarcinoma cells SPC al and GLC 82.