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目的探讨特异性下调葡萄糖调节蛋白78(GRP78)对涎腺腺样囊性癌(ACC)侵袭和转移能力的影响;方法应用siRNA干扰技术,特异性下调ACC-2细胞内GRP78的表达,通过划痕实验、Transwell实验、四甲基偶氮唑盐(MTT)实验和细胞骨架染色,观察下调GRP78表达后细胞的变化,并应用免疫印迹法检测下调GRP78表达对基质金属蛋白酶(MMP)-2、MMP-9、基质金属蛋白酶抑制因子(TIMP)-1和TIMP-2的影响。结果划痕实验显示,下调GRP78的表达能够抑制划痕愈合;Transwell实验发现,下调GRP78在ACC-2的表达明显降低穿过的细胞数;下调GRP78表达后细胞骨架染色发现,骨架纤维明显增粗;MTT实验显示,下调GRP78后细胞的增殖能力降低1/3;免疫印迹法检测结果发现,下调GRP78表达后ACC-2细胞的MMP-2、MMP-9的表达降低,TIMP-1、TIMP-2的表达增高。结论特异性下调GRP78的表达能够降低ACC的侵袭和转移能力,这与GRP78参与调节MMP和TIMP的表达有关。
Objective To investigate the effects of GRP78 on the invasion and metastasis of human salivary adenoid cystic carcinoma (ACC) .Methods The expression of GRP78 in ACC-2 cells was downregulated by siRNA interference. The changes of GRP78 expression were observed by Transwell assay, MTT assay and cytoskeleton staining. Western blotting was used to detect the expression of GRP78 on the expression of matrix metalloproteinase (MMP) -2, MMP-9, Matrix Metalloproteinase Inhibitors (TIMP) -1 and TIMP-2. Results Scratch experiments showed that downregulation of GRP78 could inhibit the healing of scratches. Transwell assay showed that down-regulation of GRP78 expression in ACC-2 significantly reduced the number of cells passing through. Scaffold staining of GRP78 down-regulated GRP78 expression, ; The results of MTT assay showed that the proliferation ability of GRP78 cells decreased by 1/3 after immunostaining. The results of Western blotting showed that the expression of MMP-2 and MMP-9 in ACC-2 cells decreased after GRP78 downregulation, while TIMP-1 and TIMP- 2 expression increased. Conclusion The down-regulation of GRP78 expression can reduce the invasion and metastasis of ACC, which is related to the regulation of GRP78 on the expression of MMP and TIMP.