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The Effect of Tissue Factor on the Invasive Ability of Human Gastric Cancer Cells Line SGC7901 in Vitro

Author: MaHaiTao
Tutor: CaoNong
School: Lanzhou University
Course: General Surgery
Keywords: Tissue factor Gastric cancer cells Eukaryotic expression vector Invasion force Nested PCR
CLC: R735.2
Type: Master's thesis
Year: 2010
Downloads: 14
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Abstract


The aim of the present study was designed to build human gastric cancer cell line stably transfected with human tissue factor (TF) (SGC7901) TF preliminary study of gastric cancer cells in vitro invasion , and to provide an experimental basis to further explore the mechanism of metastasis of gastric cancer cells . Method 1 using nested PCR from human placental tissue to clone TFcDNA, and construct a recombinant human tissue factor eukaryotic expression vector TF- pcDNA3.1 . The application of liposome -mediated transfection into human gastric cancer cell line SGC7901, stable expression of TF cells using G418 selection . 2 using flow cytometry and RT-PCR analysis of the expression levels of TF . 3 , respectively, the Transwell experiments to observe changes in the ability of the cells in vitro invasion . Results The recombinant plasmid TF-pcDNA3.1 Xba I / Hind Ⅲ digestion after a 1.2% agarose gel electrophoresis , about 5.4kb and 900bp 2 , fragment , with the expected results . Completed by the Technical Services Ltd. of Shanghai Sangon gene sequencing , in line with TF cDNA sequence , indicating that the vector was successfully constructed . 2 the stably transfected success SGC7901/TF cells within TFmRNA expression levels were significantly higher in the β-actin expression was no significant difference in the case of the experimental group (4.28 ± 0.37) was significantly higher than the negative control (1.12 ± 0.32) and blank control group ( 0.99 ± 0.29 ) ( F = 97.2 , P lt ; 0.01 ) 3 SGC7901/TF cell surface TF antigen (66.12% ± 5.09%) was significantly higher with the negative control ( 10.75 % ± 1.49 % ) and blank group ( 10.31 % ± 1.83%) there were significant differences ( F = 294.3 , P lt ; 0.01) . 4 negative controls (38.33 ± 2.52) and the control group (38.00 ± 2.65) compared to the experimental group (48.67 ± 2.52) cells in vitro invasion force enhancement (F = 16.8, P lt; 0.01). Conclusion 1 gene sequencing confirmed the success of this study to build a stable and efficient expression of TF in human gastric cancer cell lines SGC7901/TF . 2 Transwell experiments confirmed preliminary TF can enhance the invasion ability of gastric cancer cells in vitro . The 3 people gastric cancer cell lines SGC7901/TF experimental basis for further study of the role of TF in tumor metastasis .

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Gastric neoplasms
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