Effect of Recombinant Tissue Factor Pathway Inhibitor 2 Gene on The Invasion of Human Pancreatic Cancer in Vitro and in Vivo
LI Jian-shen
Abstract
LI Jian-shen
Abstract
Objective To investigate the invasion ability of Panc-1 cells in vivo and in vitro after being transfected with tissue factor pathway inhibitor 2 gene(TFPI-2).Methods The expression vector pEGFP-C1-TFPI-2 was transfected into human pancreatic cancer line Panc-1 cells by using liposome.TFPI-2 mRNA and protein of transfected and nontransfected cells were detected by reverse transcription-polymerase chain reaction(RT-PCR)and Western blot respectively.The tumor cells invasive behavior of transfected(Panc-1-TFPI-2)and nontransfected(Panc-1-V and Panc-1-P)cells were assessed in vitro through Boyden Chamber method.The transfected and nontransfected cells were implanted into nude mice to observe its growth and metastasis in vivo.Results Expressions of mRNA and protein of TFPI-2 were confirmed in transfected cells.After TFPI-2 transfection,the number of Panc-1-TFPI-2,Panc-1-V and Panc-1-P cells passing through membrane of Boyden Chamber were 24.4±3.5,61.3±4.1 and 60.2±3.9,respectively.The number of TFPI-2-expressing cells to traverse a Matrigel-coated membrane was obviously decreased compared with that of non-expressing cells,the invasion ability was lower than that before transfection in vitro.The subcutaneous tumor volume of the Panc-1-TFPI-2 group was(438.0±69.8)mm3,the Panc-1-V group was(852.0±102.9)mm3 and the Panc-1-P group was(831.0±78.1)mm3,P0.05.The metastasis to liver and lung and muscular invasion occurred in the Panc-1-V group and the Panc-1-P group.There were no muscular invasion and metastatic lesions in the Panc-1-TFPI-2 group.Conclusion TFPI-2 gene expression may obviously inhibit the invasion ability of pancreatic cancer cells in vitro and in vivo,which provides an experimental basis for the treatment of human pancreatic cancer by gene therapy.
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Objective To investigate the invasion ability of Panc-1 cells in vivo and in vitro after being transfected with tissue factor pathway inhibitor 2 gene(TFPI-2).Methods The expression vector pEGFP-C1-TFPI-2 was transfected into human pancreatic cancer line Panc-1 cells by using liposome.TFPI-2 mRNA and protein of transfected and nontransfected cells were detected by reverse transcription-polymerase chain reaction(RT-PCR)and Western blot respectively.The tumor cells invasive behavior of transfected(Panc-1-TFPI-2)and nontransfected(Panc-1-V and Panc-1-P)cells were assessed in vitro through Boyden Chamber method.The transfected and nontransfected cells were implanted into nude mice to observe its growth and metastasis in vivo.Results Expressions of mRNA and protein of TFPI-2 were confirmed in transfected cells.After TFPI-2 transfection,the number of Panc-1-TFPI-2,Panc-1-V and Panc-1-P cells passing through membrane of Boyden Chamber were 24.4±3.5,61.3±4.1 and 60.2±3.9,respectively.The number of TFPI-2-expressing cells to traverse a Matrigel-coated membrane was obviously decreased compared with that of non-expressing cells,the invasion ability was lower than that before transfection in vitro.The subcutaneous tumor volume of the Panc-1-TFPI-2 group was(438.0±69.8)mm3,the Panc-1-V group was(852.0±102.9)mm3 and the Panc-1-P group was(831.0±78.1)mm3,P0.05.The metastasis to liver and lung and muscular invasion occurred in the Panc-1-V group and the Panc-1-P group.There were no muscular invasion and metastatic lesions in the Panc-1-TFPI-2 group.Conclusion TFPI-2 gene expression may obviously inhibit the invasion ability of pancreatic cancer cells in vitro and in vivo,which provides an experimental basis for the treatment of human pancreatic cancer by gene therapy.
Key concepts: Transfection, In vivo, Molecular biology, Matrigel, Tissue factor pathway inhibitor, In vitro, Biology, Gentamicin protection assay