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Inhibition of CD147 Gene Expression Via RNA Interference Reduces Tumor Cell Invasion, Tumorigenicity and Increases Chemosensitivity to Gemcitabine in Pancreatic Cancer Cell Line Panc-1 Cells

Author: QuLiLi
Tutor: WangShuKui
School: Nanjing Medical University
Course: Clinical Laboratory Science
Keywords: CD147 shRNA Panc - 1 Proliferation Attack
CLC: R735.9
Type: Master's thesis
Year: 2011
Downloads: 18
Quote: 0
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Abstract


Objective: To investigate CD147 shRNA transfection of human pancreatic cancer cells Panc - 1 CD147 expression levels and PANC - 1 cells secrete MMPs , proliferation, invasion and resistance , and to further explore the impact of tumor growth in vivo . Methods: 1. Laboratory previously constructed recombinant plasmid pSilencer-CD147shRNA stable transfected into PANC - 1 cells , the use of quantitative PCR and Western blotting assay transfected cells CD147 expression levels change ; 2 . MTT assay detection transfected pSilencer-CD147shRNA 24, 48 and 72h after Panc-1 cell proliferation levels and the chemotherapy drug gemcitabine gemcitabine sensitivity changes ; 3 gelatin zymography transfection the pSilencer-CD147shRNA PANC - 1 cells MMP - 2 , MMP-9 activity changes ; 4 . Transwell method to detect the transfection the pSilencer-CD147shRNA Panc-1 cells in vitro invasive ; 5. using nude mice xenograft model observed transfection of pSilencer-CD147shRNA on Panc - 1 cells in vivo tumor growth and immunohistochemical analysis of tumor tissue CD147 expression level differences . Results: 1 . Transfected with pSilencer-CD147shRNA , cells Panc - 1 CD147 mRNA and protein expression level was significantly reduced , two for point of CD147 different bit of shRNA interference effect , respectively, to 80.81% ( P lt ; 0.05 ) and 34.58 % ( P lt ; 0.05 ) , and select the interference better cell clones to establish stable transfection strains for the study of the following ; 2.CD147 shRNA PANC - 1 cells was significantly decreased to 24, 48 and 72 h after transfection, the proliferation of 69.99% (P lt; 0.05), 70.95% (P lt; 0.05), 70.61% (P lt; 0.05); and the chemotherapy drug gemcitabine he Bin sensitivity increased ( P lt ; 0.05 ) ; 3 . Expressions of CD147 shRNA transfer dye Panc -1 cells of MMP -2 , MMP - 9 level decreased ( P lt ; 0.05 ) ; CD147 shRNA transfected PANC - 1 cells in vitro invasion capacity decreased ( P lt ; 0.05 ) ; CD147 shRNA nude the subcutaneous tumor size decreased to 27.86 % ( P lt ; 0.05 ) ; immunohistochemical analysis of tumor tissue expression levels of CD147 reduced . Conclusion: 1. Successfully screened CD147 shRNA interference better stable transfection lines ; CD147 shRNA can down-regulate the expression of CD147 level , and to reduce the proliferation of pancreatic cancer , invasive ability and improve the sensitivity of the chemotherapy drug gemcitabine gemcitabine .

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Pancreatic tumors
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