miR-582-5p在唾液腺腺样囊性癌侵袭、转移中的作用

Micro RNA-582-5p regulates invasion and migration of salivary gland adenoid cystic carcinoma

目的 :探讨micro RNA-582-5p(miR-582-5p)对唾液腺腺样囊性癌侵袭、转移能力的调控作用。方法 :通过q PCR验证腮腺腺样囊性癌与正常组织中miR-582-5p的表达水平差异,以唾液腺腺样囊性癌肺转移细胞株SACC-83为实验对象,通过脂质体介导,将miR-582-5p的拟似物(miR-582-5p mimics)转染至SACC-83细胞,升高miR-582-5p的表达水平。采用荧光实时定量PCR(RT-PCR)检测转染前、后SACC-83中miR-582-5p的表达变化,通过Transwell实验检测细胞侵袭、迁移能力的改变,通过荧光素酶报告实验验证靶基因,Western印迹检测miR-582-5p的靶基因整合素FOXC1的表达变化,免疫组织化学检测FOXC1在癌与正常组织中的差异表达,应用SPSS 16.0软件包对所得数据进行t检验或单因素方差分析。结果:miR-582-5p在唾液腺腺样囊性癌组织中低表达,在正常组织中高表达。转染miR-582-5p mimics后的SACC-83中,miR-582-5p的表达显著上调(P<0.001),SACC-83细胞的侵袭迁移能力显著降低(P<0.001)。Western印迹检测结果显示,在SACC-83中升高miR-582-5p的表达水平后,FOXC1表达显著降低。FOXC1在唾液腺腺样囊性癌组织中高表达,在正常组织中低表达。结论 :低表达miR-582-5p有助于维持腺样囊性癌的侵袭、转移特性;调高其表达水平,能有效抑制SACC-83的侵袭、迁移能力。miR-582-5p可能通过调控其靶基因FOXC1的表达而发挥作用。

PURPOSE： To investigate the function of miR-582-5p in cell invasion and migration in salivary gland adenoid cystic carcinoma （ACC）. METHODS： The expression levels of miR-582-Sp were detected by real-time PCR in parotid ACC cancers and normal tissues. Cell invasion and migration were detected by Transwell assays. Western blotting and IHC were performed to detect the expression of FOXC1, which was predicted as target gene of miR-582-Sp. Student＇s t test or one-way ANOVA were performed to analyze the data using SPSS 16.0 software package. RESULTS： After transfection with miR- 582-5p mimics, the expression of miR-582-5p was significantly up-regulated （P〈0.001）. The invasion and migration ability of SACC-83 was substantially inhibited by miR-582-5p.The target gene of miRNA was identified by luciferase reporter assay. Western blotting showed that FOXC1 was positive in SACC-83. With the elevated expression of miR-582-Sp, the expression of FOXC1 was down regulated substantially in SACC-83. FOXC1 was found with higher expression in SACC compared with adjacent normal tissues. CONCLUSIONS： Lower expression of miR-582-5p played an important role in the process of invasion and migration of SACC-83. Overexpression of miR-582-5p in SACC-83 could＇ substantially suppress the ability of cell invasion and migration. MiR-582-Sp may exert its action by regulating the expression of target gene FOXC 1.