AFAP-1L2对胰腺癌细胞侵袭及转移的影响及机制
| 作者: |
1刘博,
1戚诚,
2刘学臣,
1赵晓东
1 河北医科大学第二医院 肿瘤外科，河北 石家庄 050000 2 河北医科大学第二医院 消化内科，河北 石家庄 050000 |
| 通讯: |
赵晓东
Email: larrygh@aliyun.com |
| DOI: | 10.3978/.10.3978/j.issn.1005-6947.2015.09.010 |
摘要
目的：探讨AFAP-1L2表达下调对胰腺癌细胞的影响及其机制。
方法：人胰腺癌SW1990细胞分别转染靶向AFAP-1L2的干扰质粒siAFAP-1L2和阴性对照siRNA，以未处理的SW1990细胞为空白对照，检测各组细胞的迁移与侵袭能力，以及肿瘤浸润相关分子的蛋白与mRNA变化；免疫共沉淀法检测AFAP-1L2蛋白与p85α蛋白相互作用关系。
结果：与空白对照SW1990细胞比较，转染siAFAP-1L2下调AFAP-1L2后，SW1990细胞的迁移与侵袭能力明显降低，p85α及α-pAkt表达降低，α-Akt表达升高（均P<0.05），MMP-9与E-cadherin表达无变化（均P>0.05）；转染阴性对照siRNA的SW1990细胞各项指标无明显变化（均P>0.05）。免疫共沉淀显示，胰腺癌SW1990细胞中AFAP-1L2蛋白与p85α蛋白存在相互作用。
结论：AFAP-1L2可能通过与p85α相互作用调控PI3K/Akt通路，从而影响胰腺癌细胞的迁移及浸润，下调AFAP-1L2表达能抑制胰腺癌细胞迁移与侵袭能力。
关键词：
胰腺肿瘤；肌动蛋白丝相关蛋白1相似蛋白2；肿瘤侵润
方法：人胰腺癌SW1990细胞分别转染靶向AFAP-1L2的干扰质粒siAFAP-1L2和阴性对照siRNA，以未处理的SW1990细胞为空白对照，检测各组细胞的迁移与侵袭能力，以及肿瘤浸润相关分子的蛋白与mRNA变化；免疫共沉淀法检测AFAP-1L2蛋白与p85α蛋白相互作用关系。
结果：与空白对照SW1990细胞比较，转染siAFAP-1L2下调AFAP-1L2后，SW1990细胞的迁移与侵袭能力明显降低，p85α及α-pAkt表达降低，α-Akt表达升高（均P<0.05），MMP-9与E-cadherin表达无变化（均P>0.05）；转染阴性对照siRNA的SW1990细胞各项指标无明显变化（均P>0.05）。免疫共沉淀显示，胰腺癌SW1990细胞中AFAP-1L2蛋白与p85α蛋白存在相互作用。
结论：AFAP-1L2可能通过与p85α相互作用调控PI3K/Akt通路，从而影响胰腺癌细胞的迁移及浸润，下调AFAP-1L2表达能抑制胰腺癌细胞迁移与侵袭能力。
Influence of AFAP-1L2 on invasion and metastasis of pancreatic cancer cells and the mechanism
CorrespondingAuthor:ZHAO Xiaodong Email: larrygh@aliyun.com
Abstract
Objective: To investigate the influence of AFAP-1L2 down-regulation on pancreatic cancer cells and its mechanism.
Methods: Human pancreatic cancer SW1990 cells were transfected with AFAP-1L2 targeting plasmids siAFAP-1L2 or negative control siRNA sequences, using untreated SW1990 cells as blank control, and then the migration and invasion ability, and the protein and mRNA levels of the molecules related to metastasis and invasion in each group of cells were detected. The interaction between AFAP-1L2 and p85α protein was tested by co-immunoprecipitation assay.
Results: Compared with the blank control SW1990 cells, in SW1990 cells with down-regulated AFAP-1L2 expression after siAFAP-1L2 transfection, the migration and invasion ability was decreased, expressions of p85α and α-pAkt were decreased, and α-Akt expression was increased (all P<0.05), while the expressions of MMP-9 and E-cadherin showed no significant change (both P>0.05); all the studied parameters in SW1990 cells transfected with negative control siRNA seqeunces had no obvious change (all P>0.05). Co-immunoprecipitation assay showed that there was interaction between AFAP-1L2 and p85α protein in SW1990 cells.
Conclusion: AFAP-1L2 may regulate PI3K/Akt pathway through interaction with p85α and thereby influence the migration and invasion of pancreatic cancer cells, and down-regulating AFAP-1L2 expression can decrease the migration and invasion ability of pancreatic cancer cells.
Keywords：
Methods: Human pancreatic cancer SW1990 cells were transfected with AFAP-1L2 targeting plasmids siAFAP-1L2 or negative control siRNA sequences, using untreated SW1990 cells as blank control, and then the migration and invasion ability, and the protein and mRNA levels of the molecules related to metastasis and invasion in each group of cells were detected. The interaction between AFAP-1L2 and p85α protein was tested by co-immunoprecipitation assay.
Results: Compared with the blank control SW1990 cells, in SW1990 cells with down-regulated AFAP-1L2 expression after siAFAP-1L2 transfection, the migration and invasion ability was decreased, expressions of p85α and α-pAkt were decreased, and α-Akt expression was increased (all P<0.05), while the expressions of MMP-9 and E-cadherin showed no significant change (both P>0.05); all the studied parameters in SW1990 cells transfected with negative control siRNA seqeunces had no obvious change (all P>0.05). Co-immunoprecipitation assay showed that there was interaction between AFAP-1L2 and p85α protein in SW1990 cells.
Conclusion: AFAP-1L2 may regulate PI3K/Akt pathway through interaction with p85α and thereby influence the migration and invasion of pancreatic cancer cells, and down-regulating AFAP-1L2 expression can decrease the migration and invasion ability of pancreatic cancer cells.