沉默LIMK1促进DADS抑制SW480细胞迁移与侵袭  

Promotion of silence LIMK1 on diallyl disulfide inhibiting the migration and invasion of human colon cancer SW480 cells

作　　者：苏坚[1,2] 周钰娟[2] 廖前进[2] 史玲[2] 夏红[2] 苏琦[2] 

机构地区：[1]南华大学附属二医院病理科,湖南衡阳421001 [2]南华大学肿瘤研究所,湖南省高校肿瘤细胞与分子病理学重点实验室,湖南衡阳421001

出　　处：《中国药理学通报》2013年第6期760-766,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 31000629,N081102854);湖南省卫生厅科研计划项目资助项目(NoB2008-067,B2010-038)

摘　　要：目的探讨LIMK1沉默对DADS抑制SW480细胞迁移与侵袭的影响。方法 RNA干扰技术建立稳定LIMK1-miRNA/SW480细胞株;免疫组化和Western blot检测DADS对沉默LIMK1结肠癌SW480细胞LIMK1和磷酸化LIMK1蛋白表达的影响。划痕实验和侵袭实验检测LIMK1 RNA沉默与DADS对SW480细胞迁移与侵袭的影响。结果 RT-PCR与Western blot显示,LIMK1-miR/SW480细胞LIMK1mNRA与蛋白表达明显下调,表明成功构建稳定沉默LIMK1基因的SW480细胞株。免疫组化和Western blot显示,45 mg.L-1DADS处理和沉默LIMK1组较未转染组与空载体组SW480细胞LIMK1蛋白和磷酸化LIMK1明显下调(P<0.05)。划痕实验和侵袭实验发现,沉默LIMK1或DADS处理SW480细胞的迁移与侵袭能力较未转染组与空载体组明显抑制(P<0.05)。而DADS处理沉默组抑制SW480细胞迁移和侵袭能力更为明显(P<0.05)。结论沉默LIMK1基因可抑制SW480细胞迁移与侵袭,增强DADS抑制SW480细胞迁移与侵袭作用。Aim To investigate the effects of silence LIMK1 on diallyl disulfide （DADS） inhibiting the mi- gration and invasion of human colon cancer SW480 cells. Methods The stable low expression of LIMK1 ceils （LIMKl-miR/SW480） was established by mi- croRNA interference in SW480 cell line. The effects of DADS on the expression of LIMK1 and p-LIMK1 in LIMKl-miR/SW480 cells were detected by immunocy- tochemistry and Western blot. Migration and invasion potentials were examined by scratch test and transwell membrane assays. Results RT-PCR and Western blot showed that the expression of LIMK1 mRNA and pro- tein was significantly downregulated in LIMKI-miR/ SW480, which indicated the stable low expression of LIMK1 cells was successfully established by microRNA interference. The expression of LIMK1 and p-LIMK1 in the silence LIMK1 group and treatment group with45 mg .L-1 DADS compared to the no-transfection and vector groups was significantly downregulated, which was observed by Western blot and immunocyto- chemistry. Scratch test and transwell membrane assay showed that the ability of migration and invasion was obviously reduced by silence LIMK1 or treatment with DADS （P 〈0.05）. Especially, the migration and in- vasion of SW480 cells were inhibited markedly in si- lence LIMK1 and simultaneously treatment with DADS （P 〈 0.05）. Conclusion Silence LIMK1 gene could inhibit the migration and invasion of SW480 cells and enhance the inhibition of DADS on the migration and invasion of SW480 cells.

关 键 词：二烯丙基二硫 结肠癌SW480细胞 LIMK1 RNA干扰 迁移与侵袭 

分 类 号：R284.1[医药卫生—中药学] R342.2[医药卫生—中医学]