| Objective: To investigate the expression of KAI1 protein and mRNA in gastric carcinoma cell lines with different degrees of differentiation,and to compare the differences in growth and migration ability of different gastric carcinoma cell lines.Methods: 1.Three kinds of gastric carcinoma cells were selected: the poorly differentiated human gastric carcinoma cell line BGC823,the moderately differentiated human gastric carcinoma cell line SGC7901,and the highly differentiated human gastric carcinoma cell line MKN28.To detect the protein and mRNA expression of the KAI1 in selected gastric carcinoma cells by Western Blot and RT-PCR method.2.Tetrazolium salt colorimetry(MTT)was used to detect the growth of three kinds of gastric carcinoma cells.3.Flow cytometry was used to detect the difference in cell cycle of three gastric carcinoma cell lines.4.Transwell assay to detect cell migration in three gastric carcinoma cell lines.5.Fifteen female nude mice aged 4-6 weeks were randomly divided into three groups.According to the type of cells inoculated,they were divided into BGC823 group,SGC7901 group and MKN28 group nude mice.The spirit,diet and abdomen of nude mice were observed daily,and the abdominal circumference of nude mice was measured every 3 days,and plot the abdominal growth curve.6.After 4 weeks,the nude mice were sacrificed by cervical dislocation,and the tumorigenesis in the organs of the nude mice was observed,and the tumorigenesis rate was calculated,as well as the number and weight of mesenteric xenografts in the nude mice.The organs were examined by histopathology.Results: 1.The relative expressions of KAI1 protein and mRNA in human gastric carcinoma BGC823 cells,SGC7901 cells and MKN28 cells increased in turn,and the difference was statistically significant(p<0.05).2.The relative proliferation rate of BGC823,SGC7901 and MKN28 cells in 24 h human gastric carcinoma showed a decreasing trend,and the difference between the two groups was statistically significant(p<0.05).The relative proliferation rate of human gastric carcinoma MKN28 cells at 48 h was lower than that of human gastric carcinoma BGC823 and SGC7901 cells,and the difference was statistically significant(p<0.05).3.The proportion of S phase in human gastric carcinoma BGC823,SGC7901,and MKN28 cells also decreased gradually(p<0.05),and human gastric carcinoma BGC823 cells had the highest proportion of S phase,the most DNA synthesis,and the strongest proliferation ability.4.The number of migration of human gastric carcinoma BGC823,SGC7901,MKN28 cells decreased in turn,and the difference between the three groups was statistically significant(p<0.05).5.According to the curve of abdominal circumference growth,the abdominal circumference of nude mice in the BGC823 group increased faster than that of SGC7901 group and MKN28 group,and the maximum abdominal circumference of nude mice in the three groups was compared in pairs,the difference was statistically significant(p<0.05).6.Comparison of three groups of nude mice xenografts: The number of organs in the BGC823 group was the highest,mainly in the mesentery,omentum,peritoneum,stomach,liver,pancreas,spleen,kidney,and diaphragm.The MKN28 group was the least,mainly the mesentery and omentum,and the number of metastatic organs between the three groups were statistically significant(p<0.05).The number and weight of mesenteric tumors in BGC823 group,SGC7901 group and MKN28 group showed a decreasing trend,and the difference between the two groups was statistically significant(p<0.05).Conclusion:1.KAI1 protein and mRNA expression levels were negatively correlated with gastric carcinoma cell lines differentiation;2.Gastric carcinoma cell lines with high expression of KAI1 have weak growth and migration ability in vitro;while gastric carcinoma cell lines with low expression of KAI1 have strong growth and migration ability;3.A nude mice model of abdominal xenograft was successfully constructed,and nude mice with KAI1 high expression of gastric carcinoma cell line were inoculated.The growth and metastasis ability of intraperitoneal xenografts in vivo was weak.Conversely,nude mice inoculated with KAI1 low expression of gastric carcinoma cell line were used for growth and metastasis of abdominal transplanted tumor strong ability;4.KAI1 protein and mRNA expression levels may negatively regulate the growth and migration of gastric carcinoma cell lines. |
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