RAB5A反义RNA对人肺腺癌细胞系侵袭转移影响的体外研究

目的　已知RAB5A基因过表达与人非小细胞肺癌恶性表型形成相关,本研究在此基础上进一步探 讨该基因过表达在人肺腺癌恶性演进中的作用。　方法　将RAB5A反义表达载体稳定转染入高浸润人肺腺癌细 胞系L18和高转移人肺腺癌细胞系95D中,采用重组基底膜侵袭、与基底膜成分黏附能力测定、趋化运动能力测 定、明胶酶分泌与活性检测等体外实验方法观察转染前后细胞生物学特性的改变。　结果　RAB5A反义RNA稳 定转染后L18细胞趋化运动和侵袭基底膜能力显著降低(P<0.05),明胶酶活性检测发现转染后细胞MMP 2的分 泌能力明显减弱;稳定转染后95D细胞趋化运动和侵袭基膜能力显著降低(P<0.05)。　结论　体外实验显示, RAB5A基因过表达对肿瘤细胞的趋化运动能力和侵袭重组基底膜能力起重要作用,进一步提示RAB5A基因过表 达在人肺腺癌的侵袭转移过程中发挥一定作用。

STUDY ON THE EFFECT OF ANTISENSE RNA OF RAB5A ON INVASION AND METASTASIS OF HUMAN LUNG ADENOCARCINOMA CANCER IN VITRO

Objective It is indicated that over-expression of RAB5A gene is associated with neoplasia metastasis. In this study we did some research work to determine the relation of the over-expression of RAB5A gene to the malignancy progression of human lung adenocarcinoma cancer in vitro. Methods We transfected RAB5A antisense construct into human lung adenocarcinoma cell line L18 which manifested higher invasive capability and 95D with higher metastatic potential. The changes of biological characters between non-transfected and transfected cells were observed, by using reconstituted basement membrane invasion technique, analysis of adhesion capability of tumor cell, assay of chemotactic migration of tumor cells and analysis of gelatinase SDS-PAGE. Results The results showed that the capability of adhesion of transfected Ll8 and 95D had no change, the capability of invasion of basement membrane, chemotactic migration transfected L18 and 95D cells were lower, the secreting activity of gelatinase MMP2 of transfected L18 decreased obviously.Conclusion The results in vitro suggested that the over-expression of RAB5A gene play important roles in chemotactic migration and invasion of basement membrane, which may be involved in invasion and metastasis of neoplasma.