真核表达的人Y盒结合蛋白1(YB-1)促进人HepG2肝癌细胞的增殖和迁移

Eukaryotic expression of human Y-box binding protein 1(YB-1)promotes the proliferation and migration of human HepG2 cells

目的构建带有FLAG标签的Y盒结合蛋白1(YB-1)真核表达载体,转染至HepG2肝癌细胞探究YB-1对肝癌细胞增殖和迁移的影响。方法以人卵巢文库为模板并采用PCR扩增出YB-1基因片段,经双酶切后与pcDNA3.0-FLAG载体连接,构建pcDNA3.0-FLAG-YB-1真核表达载体;将其转染至HepG2细胞,通过Western blot法验证YB-1在HepG2细胞的表达;CCK-8法和集落形成实验验证YB-1对HepG2细胞生长的影响;划痕实验验证YB-1对HepG2细胞迁移的影响。结果成功构建出pcDNA3.0-FLAG-YB-1真核重组表达载体,且能在HepG2细胞中表达YB-1蛋白;YB-1能够促进HepG2细胞的增殖和迁移。结论YB-1促进HepG2细胞的增殖和迁移。

Objective To establish the eukaryotic expression vector of Y-box-binding protein 1(YB-1) with FLAG-tagged and transfect it into hepatocellular carcinoma HepG2 cells to identify the effects of YB-1 on the proliferation and migration. Methods Human YB-1 gene was amplified from the human ovary library by PCR. YB-1 fraction was double enzyme digested and connected with pcDNA3.0-FLAG vector to construct eukaryotic expression vector pcDNA3.0-FlAG-YB-1, which was transfected into HepG2 cells. The expression of YB-1 was detected by Western blotting, and the effect of YB-1 on the proliferation of HepG2 cells was determined by CCK-8 assay and clone formation. The effect of YB-1 on the migration of HepG2 cells was analyzed by wound healing assays. Results The eukaryotic expression vector pcDNA3.0-FLAG-YB-1 was successfully established. YB-1 protein can be expressed in HepG2 cells, and YB-1 promoted the proliferation and migration of HepG2 cells. Conclusion YB-1 promotes the proliferation and migration of HepG2 cells.