沉默LIMK1促进DADS抑制SW480细胞迁移与侵袭

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。

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

Aim To investigate the effects of silence LIMK1 on diallyl disulfide(DADS) inhibiting the migration and invasion of human colon cancer SW480 cells.Methods The stable low expression of LIMK1 cells(LIMK1-miR/SW480) was established by microRNA interference in SW480 cell line.The effects of DADS on the expression of LIMK1 and p-LIMK1 in LIMK1-miR/SW480 cells were detected by immunocytochemistry 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 protein was significantly downregulated in LIMK1-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 with 45 mg· L-1 DADS compared to the no-transfection and vector groups was significantly downregulated,which was observed by Western blot and immunocytochemistry.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 invasion of SW480 cells were inhibited markedly in silence 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.