Maspin在非小细胞肺癌A549细胞生物学行为中的作用

Roles of maspin in biological behaviors of A549 cells

目的:探讨乳腺丝氨酸蛋白酶抑制物(maspin)对于非小细胞肺癌细胞生物学有何影响及其作用机制。方法:PCR扩增人类maspin编码区全长,并构建maspin过表达载体,转染A549细胞。药物筛选出稳定高表达maspin蛋白的细胞株,并通过real-time PCR和Western blot鉴定。通过x Celligence系统实时动态观测转染后A549细胞的生长、迁移和侵袭情况。利用Western blot和real-time PCR的方法对此细胞生物学变化的相关基因进行分析,探讨相应的作用机制。结果:成功构建过表达Maspin的载体MSCV-maspin,并获得稳定高表达maspin的A549细胞株(A549-maspin)。A549-control细胞和A549-maspin细胞生长曲线无差异,但是迁移和侵袭曲线有显著差异。在A549-maspin细胞中integrinβ1的表达水平低于A549-control细胞。结论:Maspin对于A549细胞的生长无影响,但是抑制了A549细胞的迁移和侵袭能力,并且此抑制作用与integrinβ1相关。

AIM：To investigate the roles of maspin in the biological behaviors of non-small-cell lung cancer A549 cells.METHODS：Full-length coding region of maspin was amplified by PCR from human normal breast tissue and cloned into MSCV vector .Virus supernatants was produced by Phoenix A packaging system , and target cell line was infec-ted with the virus supernatants .The transfected cells were screened with puromycin .The cell line with maspin over-expres-sion was identified by real-time PCR and Western blot.Cell growth, migration and invasion were investigated by xCelli-gence system .RESULTS：Maspin over-expression vector was successfully constructed .The cell line with maspin over-ex-pression was established .No difference of the growth between A 549-control cells and A549-maspin cells was observed .The migration and invasion were quite different between A 549-control cells and A549-maspin cells.The expression level of inte-grin β1 in A549-maspin cells was decreased compared with A 549-control cells .CONCLUSION：Maspin has no effect on the growth of A549 cells.Maspin suppresses the migration and invasion of A 549 cells, in which integrin β1 is involved.