中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
Chinese Journal of Pathophysiology
2015年
9期
1537-1544
,共8页
周珺%周鹏%秦华龙%国风
週珺%週鵬%秦華龍%國風
주군%주붕%진화룡%국풍
非小细胞肺癌%乳腺丝氨酸蛋白酶抑制物%细胞迁移%细胞侵袭
非小細胞肺癌%乳腺絲氨痠蛋白酶抑製物%細胞遷移%細胞侵襲
비소세포폐암%유선사안산단백매억제물%세포천이%세포침습
Non-small-cell lung cancer%Maspin%Cell migration%Cell invasion
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xCelligence 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integrin β1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integrin β1相关。
目的：探討乳腺絲氨痠蛋白酶抑製物（ maspin）對于非小細胞肺癌細胞生物學有何影響及其作用機製。方法：PCR擴增人類maspin編碼區全長，併構建maspin過錶達載體，轉染A549細胞。藥物篩選齣穩定高錶達maspin蛋白的細胞株，併通過real－time PCR和Western blot 鑒定。通過xCelligence 繫統實時動態觀測轉染後A549細胞的生長、遷移和侵襲情況。利用Western blot和real－time PCR的方法對此細胞生物學變化的相關基因進行分析，探討相應的作用機製。結果：成功構建過錶達Maspin的載體MSCV－maspin，併穫得穩定高錶達maspin的A549細胞株（ A549－maspin）。 A549－control細胞和A549－maspin細胞生長麯線無差異，但是遷移和侵襲麯線有顯著差異。在A549－maspin細胞中integrin β1的錶達水平低于A549－control細胞。結論：Maspin對于A549細胞的生長無影響，但是抑製瞭A549細胞的遷移和侵襲能力，併且此抑製作用與integrin β1相關。
목적：탐토유선사안산단백매억제물（ maspin）대우비소세포폐암세포생물학유하영향급기작용궤제。방법：PCR확증인류maspin편마구전장，병구건maspin과표체재체，전염A549세포。약물사선출은정고표체maspin단백적세포주，병통과real－time PCR화Western blot 감정。통과xCelligence 계통실시동태관측전염후A549세포적생장、천이화침습정황。이용Western blot화real－time PCR적방법대차세포생물학변화적상관기인진행분석，탐토상응적작용궤제。결과：성공구건과표체Maspin적재체MSCV－maspin，병획득은정고표체maspin적A549세포주（ A549－maspin）。 A549－control세포화A549－maspin세포생장곡선무차이，단시천이화침습곡선유현저차이。재A549－maspin세포중integrin β1적표체수평저우A549－control세포。결론：Maspin대우A549세포적생장무영향，단시억제료A549세포적천이화침습능력，병차차억제작용여integrin β1상관。
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.
