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目的探讨多肽N-乙酰氨基半乳糖转移酶2(ppGalNAc-T2)对胃癌细胞株SGC-7901细胞性能的影响。方法构建ppGalNAc-T2RNA干扰真核载体(pSilenCircle-SiT2)、正义真核表达载体(pEGFP-C1-T2)及空载体(pEGFP-C1),分别转染胃癌细胞株SGC-7901细胞。MTT法检测上述各组转染后细胞对SGC-7901细胞增殖的影响;细胞黏附和穿膜实验分别检测其黏附力和迁移力的变化;同时采用RT-PCR和Westernblot检测MMP2、MMP14、TGF-β1等与肿瘤细胞浸润、转移、增殖相关基因mRNA及蛋白的表达水平。结果转染正义真核表达载体的SGC-7901细胞,随着ppGalNAc-T2表达量的增加,细胞的生长增殖受到抑制,并对纤连蛋白、基质胶和透明质酸的黏附能力降低,但其侵袭迁移能力变化不明显;而转染ppGalNAc-T2RNA干扰载体的SGC-7901细胞,细胞生长最快。ppGalNAc-T2与肿瘤细胞相关因子TGF-β1、MMP2的mRNA表达水平成负相关,但对MMP14无明显影响,Westernblot检测相关蛋白结果与mRNA表达水平基本一致。结论ppGalNAc-T2可影响胃癌细胞株SGC-7901细胞的增殖、黏附能力,并影响与肿瘤细胞浸润、转移、增殖相关基因mRNA及蛋白的表达水平。
Objective To investigate the effect of polypeptide N-acetylgalactosamine 2 (ppGalNAc-T2) on the cell proliferation of gastric cancer cell line SGC-7901. Methods We constructed ppGalNAc-T2 RNA interference eukaryotic vector (pSilenCircle-SiT2), eukaryotic expression vector (pEGFP-C1-T2) and empty vector (pEGFP-C1) and transfected it into SGC-7901 cells. The cell proliferation and proliferation of SGC-7901 cells were detected by MTT assay. The adhesion and migration of SGC-7901 cells were detected by MTT assay. The expressions of MMP2, MMP14 and TGF- β1 and other tumor cells infiltration, metastasis, proliferation-related gene mRNA and protein expression levels. Results The SGC-7901 cells transfected with sense eukaryotic expression vector inhibited the proliferation of SGC-7901 cells and inhibited the adhesion of fibronectin, matrigel and hyaluronic acid with the increase of ppGalNAc-T2 expression. However, However, SGC-7901 cells transfected with ppGalNAc-T2RNA had the fastest cell growth. The expression of ppGalNAc-T2 was negatively correlated with the mRNA level of TGF-β1 and MMP2, but had no significant effect on MMP14. Western blot showed that the expression of ppGalNAc-T2 protein was consistent with that of mRNA. Conclusion ppGalNAc-T2 can affect the proliferation and adhesion of gastric cancer cell line SGC-7901 and affect the expression of mRNA and protein related to the infiltration, metastasis and proliferation of gastric cancer cell line SGC-7901.