摘要
目的:探讨上调或下调FMNL2表达与大肠癌细胞侵袭能力,以及该基因与Src、Talin表达的相关性.方法:将FMNL2-CT过表达重组慢病毒颗粒转染SW480和HT29大肠癌细胞,构建FMNL2干扰质粒沉默SW620大肠癌细胞,通过侵袭实验观察FMNL2表达量与细胞侵袭能力关系;Westernblot检测转染前后细胞Talin、Src表达情况,及PP1处理前后Talin、Src、FMNL2三种蛋白表达变化;荧光免疫共定位检测FMNL2、Src、Talin大肠癌细胞内定位.结果:高表达FMNL2的癌细胞侵袭能力较低表达者显著提高(51.20±8.00vs38.00±4.00,P<0.05).随着大肠癌细胞内FMNL2蛋白表达量的升高或降低,Src蛋白也上升或降低(F=15.659,P<0.05),Talin则反之(SW480F=540.595,HT29F=163.816,SW620F=125.507,P<0.01).PP1处理阻断Src通路后,FMNL2、Src蛋白含量均无明显变化,而Talin蛋白表达量随之减低.免疫共定位检测FMNL2与Src或Talin在胞浆和胞膜上存在共定位关系.结论:FMNL2可以显著促进结直肠癌细胞的侵袭能力,FMNL2处于上游参与调控Src、Talin表达,并可能通过Src间接影响Talin的表达进而参与对黏附斑转换的调控.
AIM：To study the role of FMNL2 in regulating the migration of colorectal cancer cells by overexpressing and silencing FMNL2 in these cells,and to evaluate the correlation between the expression of FMNL2 gene and that of Src and Talin.METHODS：FMNL2-expressing lentivirus was infected into SW480 and HT29 cell lines.RNAi plasmid that expresses a siRNA targeting the FMNL2 gene was designed,constructed,and transfected into SW620 cells line.In vitro invasion assay was performed to investigate the influence of FMNL2 expression on colorectal cell invasion.Western blot was used to detect the expression of FMNL2,Src and Talin in cells and to assess the effect of PP1 on Talin,Src,and FMNL2 expression.Immuno-colocalization assay wasused to analyze the interaction of FMNL2 with Src and Talin.RESULTS：Cell invasion was significantly increased in cells overexpressing FMNL2（51.20 ± 8.00 vs 38.00 ± 4.00,P 0.05）.FMNL2 expression was positively correlated with Src expression（F = 15.659,P 0.05）,but negatively correlated with Talin expression.Treatment with PP1 prominently decreased Talin expression（SW480F = 540.595,HT29F = 163.816,SW620F = 125.507,all P 0.01）,but did not change FMNL2 and Src expression.FMNL2 and Talin were co-localized in the cytoplasm,and FMNL2 and Src were colocalized in the plasma membrane.CONCLUSION：FMNL2 significantly promotes invasion of colorectal cells.FMNL2 can regulate Src and Talin expression and indirectly control the transition of focal adhesions by regulating Src.
出处
《世界华人消化杂志》
CAS
北大核心
2012年第4期289-295,共7页
World Chinese Journal of Digestology
基金
广东省自然科学基金资助项目
No.9151503101000016~~
