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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。
Objective It is known that overexpression of RAB5A is associated with the formation of malignant phenotype in human non-small cell lung cancer. Based on this study, we further investigate the role of overexpression of RAB5A in the malignant progression of human lung adenocarcinoma. Methods The antisense expression vector RAB5A was stably transfected into the human lung adenocarcinoma cell line L18 and the human lung adenocarcinoma cell line 95D. The recombinant adenovirus vector was used to detect the adhesion of basement membrane to chemotactic ability Determination, gelatinase secretion and activity test and other in vitro experimental methods to observe the changes of cell biological characteristics before and after transfection. Results After transfection with RAB5A antisense RNA, the ability of L18 cells to chemotactic and invasion to basement membrane was significantly decreased (P <0.05). The gelatinase activity assay showed that the secretion of MMP 2 in transfected cells was significantly decreased. After stable transfection of 95D cells Chemotaxis and invasion of the basilar membrane decreased significantly (P <0.05). Conclusion In vitro experiments show that RAB5A gene overexpression plays an important role in the chemotactic ability of tumor cells and invasion of recombinant basement membrane, which further suggests that RAB5A overexpression plays a role in the invasion and metastasis of human lung adenocarcinoma.