TGF-β1和LPA增强整合素β6基因启动子在肝癌细胞Hep-3B中的转录活性 外科学专业论文.docxVIP

汕头大学 汕头大学医学院硕士研究生学位论文 汕头大学医学院硕士研究生学位论文 汕头大学医学院硕士研究生学位论文 PAGE PAGE III PAGE PAGE IV 结论： 1. 整合素α vβ 6在肝细胞癌组织标本及Hep-3B细胞中呈高表达。 2. TGF-β 1和LPA可能通过结合AP-1、c-Myb、HNF-1/HNF-1B、SRF或YY1等转录因子结合位 点，从而激活整合素β 6基因的启动子区，最终诱导整合素α vβ 6在Hep-3B细胞中过度 表达，这将有望为肝细胞癌的诊断和治疗提供新的潜在治疗靶点。 关键词： 肝细胞癌；整合素α vβ 6；转化生长因子–β 1；溶血磷脂酸 ABSTRACT Objective: To investigate that whether Transforming Growth Factor-beta1 (TGF-β1) and lysophospholipids Acid (LPA) can induce the overexpression of integrin αvβ6 (Itg αvβ6) in Hep-3B, and perform the preliminary study in gene promoter regions of Itgβ6. Materials and Methods: First of all, we obtained the tissue samples from patients with hepatocellular carcinoma in hospital, and then detected the expression of Itg αvβ6 by immunohistochemistry. Second- ly，to detect the expression of Itg αvβ6 in Human hepatic cancer HYPERLINK /detail_1.asp?id=75amp;serial=TCHu106 Hep-3B and normal liver cells HL7702 with reverse transcription PCR (RT-PCR). What’s more, Hep-3B cells were treated with either TGF-β1, LPA, TGF-β1+LPA, and Itgβ6 expression were examined by real-time PCR and western blotting. At last, promoter deletions were generated from pGLB-1135 by PCR, and inserted into the pGL2-Basic. The activities of a series of Itgβ6 promoter deletion mutants were measured by Luciferase reporter gene assays, with TGF-β1 or LPA stimulated. Furthermore, using Transcrip- tional Element Search System (TESS) analysis forecast the transcription factor-binding sites in the positive regulatory region of the Itgβ6 promoter. Results: Immunohistochemical results: the Itgβ6 overexpression was present in the hepatocellular carcinoma, while absent in normal liver tissue. RT-PCR results：the Itgβ6 mRNA was detected in Hep-3B cells and was not detectable in HL-7702 cells. Real-time PCR results：TGF-β1, LPA and TGF-β1+LPA induced an increase in Itg β6 expres- sion at mRNA levels in Hep-3B cells: When compared with non-treated cells （ 1.041±0.1029 ） , TGF-β1, LPA, and