论文部分内容阅读
目的:探讨microRNA-320a(miR-320a)对唾液腺腺样囊性癌侵袭、转移能力的调控作用。方法:以唾液腺腺样囊性癌高转移细胞株ACC-M为实验样品,通过脂质体介导,将miR-320a的拟似物(miR-320a mimics)转染至ACC-M细胞,升高miR-320a的表达水平。采用荧光实时定量RT-PCR检测转染前、后ACC-M中miR-320a的表达变化,通过Transwell实验检测细胞侵袭、迁移能力的改变,并通过细胞计数试剂盒-8检测转染后细胞增殖能力的变化,流式细胞仪检测miR-320a对细胞凋亡的影响,Western印迹检测miR-320a的靶基因整合素β3(integrinβ3,ITGB3)的表达变化。应用SPSS13.0软件包对所得数据进行t检验或单因素方差分析。结果:转染miR-320amimics后的ACC-M中miR-320a的表达明显上调(P<0.001)。ACC-M细胞的侵袭迁移能力显著降低(P<0.001),而细胞的增殖和凋亡无显著变化。Western印迹检测结果显示,Integrinβ3在低转移细胞株ACC-2表达阴性,在ACC-M中高表达;升高miR-320a在ACC-M中的表达,Integrinβ3表达明显降低。结论:低表达miR-320a有助于维持ACC的侵袭转移特性,调高其表达水平能有效抑制ACC-M的侵袭迁移能力。miR-320a可能通过调控其靶基因Integrinβ3的表达而发挥作用。
Objective: To investigate the regulatory effect of microRNA-320a (miR-320a) on invasion and metastasis of salivary adenoid cystic carcinoma. Methods: ACC-M was transfected into ACC-M cells by liposome-mediated transfection of miR-320a mimics into ACC-M cell lines with high metastatic potential of salivary adenoid cystic carcinoma High miR-320a expression levels. Real-time quantitative RT-PCR was used to detect the expression of miR-320a in ACC-M before and after transfection. The changes of cell invasion and migration were detected by Transwell assay. The cell proliferation was detected by cytometry kit-8 The effect of miR-320a on apoptosis was detected by flow cytometry. The expression of integrinβ3 (ITGB3), a target gene of miR-320a, was detected by Western blotting. The SPSS 13.0 software package was used for t test or one-way analysis of variance. Results: miR-320a expression was significantly up-regulated in ACC-M transfected with miR-320amimics (P <0.001). The invasion and migration of ACC-M cells were significantly reduced (P <0.001), while there was no significant change in cell proliferation and apoptosis. Western blotting results showed that Integrinβ3 was negative in ACC-2 and ACC-M, and increased the expression of miR-320a in ACC-M. The expression of Integrinβ3 was significantly decreased. Conclusion: The low expression of miR-320a helps to maintain the invasion and metastasis of ACC, and the increased expression of miR-320a can effectively inhibit the invasion and migration of ACC-M. miR-320a may play a role by regulating the expression of its target gene Integrinβ3.