| Objective: In this study, through the preparation of human gastric cancercells SGC7901orthotopic implantation nude mouse model to observe theexpression of MRP-1, MMP-7, ZNF-139difference in the primary tumor andmetastasis of gastric cancer cells in nude mouse model of orthotopictransplantation and explore the different mechanism of tissue cells in vitrochemosensitivity difference.Methods:1.The establishment of human gastric cancer cell line SGC7901(moderately differentiated adenocarcinoma) in nude mice orthotopictransplantation tumor modelCollection logarithmic phase SGC7901cells in vitro, with the preparationof the cell suspension after nude mice subcutaneous injection until thesubcutaneous tumor growth to a diameter of about1.0cm remove, take the sizeof about2mm3in gastric carcinoma inoculated into the a nude micesubcutaneously, mouse passaged, remove the transplanted tumors passaged sixtimes in the same way, cut into1mm3size of the organization blocks thestomach anterior wall subserous orthotopic implantation of human gastriccancer orthotopic transplantation nude mouse model. Off when systemicfailure signs were sacrificed to remove the orthotopic transplantation tumor,liver, spleen, peritoneal nodules, abdominal and chest swollen lymph nodes intumor-bearing mice, each specimen is divided into two, a rapidly stored at-80℃refrigerator and stored, a formalin fixed alternate.2.By immunohistochemical method (SP) detection of ZNF-139, MMP-7,the MRP-1in each organization, expressionLearn from the formalin fixed tissue by immunohistochemical method (SP) test zinc finger protein139(ZNF139), matrix metalloproteinase7(MMP-7), multidrug resistance-associated protein1(MRP-1) in theorthotopic transplantation tumor, liver metastases, spleen metastasis,peritoneal metastasis, the expression of metastatic lymph nodes.3.By RT-PCR was monitored ZNF-139mRNA, and MMP-7mRNA,MRP-1mRNA expression in various tissuesTake frozen tissue, RNA was extracted after grinding, the determinationof the concentration, reverse transcription, DNA amplification, and get theMRP-1, MMP-7, ZNF-139target genes, application of gel electrophoresis,detection of multidrug resistance-associated protein1(MRP-1) mRNA andmatrix metalloproteinase7(MMP-7) mRNA, a zinc finger protein139(ZNF-139) mRNA in the orthotopic transplantation tumor, liver metastasesand spleen metastasis, peritoneal metastasis, lymph node metastasis in eachgroup expression.Results:1.Application OB biological glue successfully established intact tissueorthotopic metastasis model of human gastric cancer, in situ tumor was100%.Two were killed before the30nude mice were killed and the tumor-bearingsurvival time was72days and81days, the autopsy shows huge orthotopictransplantation tumor and multiple peritoneal metastasis. Be tumor-bearingmice appear thin, arched, the activity difference off the neck were dissectedand the remaining28tumor-bearing mice take the orthotopic transplantationtumor, liver, spleen, peritoneal nodules, abdominal and thoracic lymph node,HE staining and histopathological physical examination, results showed that21(75%) of mice with liver metastases,19(67.86%), spleen metastasis,24(85.71%) peritoneal metastasis,11(39.29%) had lymph node metastasis.2.ZNF-139,(2) immunohistochemical method (SP) results showed thatMMP-7, higher than the MRP-1expression in the liver metastases, peritonealnodules and lymph node metastases of orthotopic tumor tissue, statisticalanalysis showed that There are significant differences (P <0.05). Spearmancorrelation analysis showed that expression of ZNF-139in each group was positively correlated with MMP-7and MRP1.3.Application by RT-PCR test results show that ZNF-139mRNA,MMP-7mRNA, MRP-1mRNA expression in liver metastases, abdominalperitoneal nodules and transfer of lymph nodes higher than orthotopic tumortissue by t The test statistic analysis showed significant differences (P <0.05).Pearson linear correlation results show that ZNF-139mRNA expression ofMMP-7mRNA and MRP-1mRNA was positively correlated within eachgroup respectively.Conclusion:1.Human gastric cancer orthotopic nude mouse model can beapproximated simulation occurs, proliferation, invasion and metastasis ofgastric cancer in the human body biological behavior compared with othertypes of models, simple operation, high rate of tumor formation, tumor fastgrowth, high cost of feeding can be used as ideal experimental tool of gastriccancer.2.As transcriptional gene regulatory factors,2.ZNF139expression ingastric cancer and metastases may promote the expression of MRP-1, MMP-7,the differential expression of the primary gastric tumor transfer of Zaojianresistance, proliferation, invasion, metastasis such as the biological behaviorof the difference.3.ZNF139in the level of gene transcription regulation, MMP-7andMRP-1expression in the same tissue was positively correlated, and may resultin gastric cancer metastases than primary tumors, MRP-1expression increasedin gastric cancer cell drug resistance. insensitive to chemotherapy, the moresusceptible of MDR. |
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