ppGalNAc-T2对胃癌细胞株SGC-7901生物学性能的影响

Influence of ppGalNAc-T2 on Biological Proporties of Gastric Cancer Cell Line SGC-7901

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 probe how polypeptide：N-acetylgalactosaminyltransferase2 （ppGalNAc-T2） affects gastric cancer cell line SGC7901. Methods ppGalNAc-T2 RNA interferential carrier （pSi-lenCircle-SiT2）,plus sense carrier （pEGFP-C1-T2） and vacuity carrier （pEGFP-C1） was separately constructed and SGC-7901 was transfected; how the multiplication of SGC-7901 was affected by ppGalN-AcT2 was detected with MTT; the change of capability of adhesion and metastasis were gauged by cell adhesion and penetration experiment; reverse RT-PCR was adopted to detect mRNA horizontal express status of ppGalNAc-T2 and determinants correlated with tumor infiltration; Western blot was used to detect express status of related protein in dissociation liquor of tumor cells （MMP2,MMP14,TGF-β 1）. Results With the increase of express volume of ppGalNAc-T2,the cell growth was depressed and the ability of adhesion to matrigel,hyaluric acid and fibronectin drops,however,metastasis ability was not changed any way. The cell growth was increased by the pSilenCircle-SiT2-transfected SGC-7901. Of on the level of mRNA,ppGalNAc-T2 had negative correlation with TGF-β1,MMP2,but didn＇t exhibit apparent correlation with MMP14. The protein express status detected by Western blot method was in accordance with the level of mRNA. Conclusion It is possible that the tune-up and tune-down of pp-GalNAc-T2 are related with adhesive metastasis of tumor.