人多肽N-乙酰氨基半乳糖转移酶-2对胶质瘤细胞U251中MMP-2、TIMP-2 mRNA表达及细胞迁移能力的影响

目的探讨多肽N-乙酰氨基半乳糖转移酶-2（ppGalNAc-T2）对胶质瘤细胞侵袭转移的作用。方法将ppGalNAc-T2的真核表达正义载体、RNA干扰载体通过脂质体稳定转入胶质瘤细胞U251;通过荧光显微镜观察及RT-PCR检测ppGalNAc-T2的表达来确定载体正确转入细胞;细胞分为以下5组：U251转染正义载体组与空载体组、U251转染干扰载体组与对照载体组、U251未转染组,通过RT-PCR方法检测各组细胞基质金属蛋白酶（MMP）-2、基质金属蛋白酶抑制剂（TIMP）-2mRNA表达的变化;通过划痕法检测各组细胞的迁移差异。结果荧光显微镜观察及RT-PCR检测结果表明,载体成功转入细胞;RT-PCR检测结果发现,随着ppGalNAc-T2的增高,MMP-2的表达降低,而TIMP-2的表达升高（P〈0.05）;划痕结果显示,正义组细胞迁移能力随ppGalNAc-T2的上调而减弱（P〈0.05）,干扰组划痕修复较快（P〈0.05）,其他对照组相近（P〈0.05）。结论 ppGalNAc-T2的上调可抑制细胞的划痕修复,由此推测ppGalNAc-T2可能与胶质细胞的侵袭转移密切相关。

Effect of Human Polypeptide N-Acetylgalactosaminyltransferase-2 on the mRNA Expression of Matrix Metalloproteinase-2, Tissue Inhibitor of Metalloproteinase-2 and Cell Migratory Capability in Human Glioma Cells U251

Objective To probe the effect of ppGalNAc-T2 on migration and invasion in human glioma cell U251.Methods ppGalNAc-T2 sense vector and RNA interference vector were transfected stably into U251 cells by Liprofectamine 2000.Fluorescence microscope and RT-PCR were used to detect the vectors were transfected into cells successfully;There were five groups：U251 transfected with sense vector and empty vector groups,U251 transfected with interference vector and control vector groups,U251 hadn’t tranfected group.RT-PCR was used to study the variational characteristics of MMP-2 and TIMP-2 expression on mRNA level.Wound healing was used to compare the capability of migration in different group cells.Results The stable upregulation and downregulation of ppGalNAc-T2 U251 cells were established.Upregulation of ppGalNAc-T2 decreased the mRNA expression of MMP-2 and increased TIMP-2.Downregulation cells showed opposite results.ppGalNAc-T2 knockdown cells migrated slower than upregulation group cells.Conclusion ppGalNAc-T2 has a negative correlation with MMP-2 and positive correlation with TIMP-2;ppGalNAc-T2 inhibited cell migration.These suggest that ppGalNAc-T2 may has an important role in migration and invasion of tumor cells.