Objective To investigate the effect of CO2 pneumoperitoneum on the tumor cell port site implantation in laparoscopic surgery. Methods Male SpraqueDawley rats were intraperitoneally injected with gastric cancer cells (cell line SGC-7901). Continuous CO2 pneumo of 15 mm Hg or 30 mm Hg were established for 5 mins, 60 mins, 120 mins and 180 mins with the injection of different concentrations of tumor cells (104/ml, 106/ml respectively). Several samples of peritoneal washing served as positive control. All collecting dishes were incubated at 37℃ with 5% CO2 concentration for one week and then examined for the presence of tumor cell under microscope. Results After one week of incubation, some of the dishes with continuous flow of CO2 gas (5 L/min) at pneumo 30 mm Hg for 60 mins or longer demonstrated tumor growth, and all peritoneal washing samples showed tumor growth, while other dishes showed negative. Conclusion The research suggests that gastric cancer cells can cause port site implantation and the concentration of tumor cells, pneumoperitoneum pressure and duration may affect the occurrence of port site implantation. It may help to find a suitable way to prevent the port site implantation in operations.
Objective To construct the eukaryotic expressive vector of human tissue factor （TF），and to abserve the effect of TF on invasion and metastasis of gastric cancer cells line. Methods The human TF cDNA was obtained from human placenta by nest PCR, and the constructed eukaryotic expressive vector TF-pcDNA3 was transfected into SGC7901 cells by lipofectamine. Stable-transfected cells were screened by G418. The expressions of TF mRNA and protein on the cells were detected by RT-PCR and Western blot. Cell motility was assessed by using Transwell experiments and wound-healing assays. Results The eukaryotic expressive vector TF-pcDNA3 was successfully constructed and transfected into SGC7901. Compared with blank control group and negative control group, the expressions of TF mRNA and TF protein in transfection group were increased, the cell motility in vitro was enhanced. Conclusion TF can enhance the ability of invasion and metastasis of gastric cancer cells in vitro.
Objective To investigate the influence of different pressures and duration of CO2 pneumoperitoneum on the adhesive and invasive ability of gastric cancer cells based on the expressions of adhesive and invasive molecules. Methods With an artificial CO2 pneumoperitoneum model in vitro, human gastric cancer cell lines including MKN-45, SGC-7901, and MKN-28 were exposed to CO2 in different environments: 0 mm Hg （1 mm Hg=0.133 kPa）, 9 mm Hg （2 h, 4 h）, and 15 mm Hg （2 h, 4 h）. The expressions of mRNA of E-cadherin, intercellular adhesion molecule-1 (ICAM-1), matrix metalloproteinase-2 (MMP-2), and vascular endothelial growth factor-A (VEGF-A) in the different environments were measured by RT-PCR. The expressions of protein of E-cadherin and ICAM-1 in the environments of 0 mm Hg and 15 mm Hg （4 h） were measured by FCM. Results With the increase of duration or pressure, RT-PCR showed that there was a downward trend in the expression of E-cadherin mRNA as well as there were upward trends in the expressions of ICAM-1, MMP-2, and VEGF-A mRNA; FCM showed that there was a downward trend in the expression of E-cadherin protein while the expression of ICAM-1 protein showed the opposite change. But there were no obvious differences under different environment (P＞0.05). Conclusions Under low pressure (≤15 mm Hg) and short time (≤4 h) of CO2 pneumoperitoneum, the adhesive and invasive ability of gastric cancer cells could not be affected, which means that under this environment, CO2 pneumoperitoneum will not increase the possibility of neoplasm metastasis.
ObjectiveTo study the expression levels of miR-339-3p and miR-339-5p in the gastric carcinoma cell lines(SGC-7901, BGC-823, and MKN-45) and gastric surface epithelium(GES-1);detect the relationship between miR-339-3p and miR-339-5p and the gastric carcinoma cell lines in vetrio experiment through the gain of function, and further significance is suggested. MethodsSYBR greenⅠreal time PCR was performed to access the expression of miR-339-3p and miR-339-5p in different cell lines(SGC-7901, BGC-823, MKN-45, and GES-1). The expression levels of miR-339-3p and miR-339-5p were verified by real time PCR experiment again after transfecting miR-339-3p mimics and miR-339-5p mimics. After that, the changes of MKN-45 cells apoptosis and proliferation at 72 h after transfection were detected by flow cytometry and CCK-8 method. ResultsThe expression levels of miR-339-3p and miR-339-5p in gastric carcinoma cell lines(SGC-7901, BGC-823, and MKN-45) were down regulated. Compared with the control group, the apoptosis of MKN-45 cell line was significantly higher(P < 0.05), the ability of proliferation of MKN-45 cell line decreased after transfecting miR-339-3p mimics and miR-339-5p mimics within 72 hours(P < 0.01). ConclusionThe expression levels of miR-339-3p and miR-339-5p significantly decreased in the gastric carcinoma cell lines(SGC-7901, BGC-823, and MKN-45) in contrast with gastric surface epithelium. MiR-339-3p and miR-339-5p may be involved in the apoptosis and proferation of the gastric carcinoma.