The Effect Of Tissue Factor On The Invasive Ability Of Human Gastric Cancer Cells Line SGC7901 In Vitro

ObjectivesThe aim was to construct the human gastric cancer cell line (SGC7901) in stable-transfected with the human tissue factor (TF), then preliminary study the influence of TF pairs of gastric cancer cells invasiveness in vitro, and provide an experimental basis for the transfer mechanism of the gastric cancer cells.Methods1 The human TFcDNA was obtained from human placenta by nest PCR and then inserted into eukaryotic expressive vector pcDNA3.1 to obtain the TF-pcDNA3.1 recombinant. This recombinant gene was introduced into human gastric cell line SGC7901 through transfection mediated by lipofectamine. Stable-transfected cells were screened by G418.2 The TF expressive levels were detected by RT-PCR and flow cytometry.3 Transwell experiments were carried out to observe the changes in cell invasion ability in vitro respectively.Results1 The constructed TF-pcDNA3.1 recombinant was identified by double digestion with XbaⅠand HindⅢ, and yielded two fragments of 5.4kb and 900bp. It was in line with the expects.2 The TFmRNA levels in the successfully transferred SGC7901/TF cells were obviously higher. On the condition thatβ-action expressed no statistics difference, the experiment group (4.28±0.37)was obviously higher than negative control (1.12±0.32)and the blank control (0.99±0.29) (F=97.2, P<0.01).3 TF antigen expression(66.12%±5.09%) on the SGC7901/TF cell surface was obviously higher. Compared to the negative control (10.75%±1.49%) and the blank control(10.31%±1.83%),the statistics difference came out. (F= 294.3, P<0.01). 4 Compared to the negative control (38.33±2.52) and blank control (38.00±2.65), the invasiveness in vitro of the experimental group increased(48.67±2.52) (F=16.8, P<0.01).Conclusions1 We successfully built the human gastric cancer cell line SGC7901/TF which expressed TF stablely and efficiently.2 Preliminarily confirmed that TF can enhance the invasive ability of gastric cancer cells in vitro by transwell.3 It established an experimental basis for further study of the mechanism of TF in tumor metastasis.