| Background:As one of the most common malignant tumors of digestive tract,gastric cancer is characterized by low early diagnosis rate and high mortality in china.Chemotherapy is one of the main methods for the treatment of advanced gastric cancer and gastric cancer.However,the multidrug resistance of tumor has a serious impact on the efficacy of chemotherapy.Multidrug resistance of tumor is the result of many factors,and its mechanism is complex.If we can study the mechanism of multidrug resistance in gastric cancer and reverse the drug resistance,it will bring a new breakthrough in the clinical treatment of gastric cancer.Survivin and cyclin D1 are highly expressed in many tumors and drug resistance of tumor cells,while not reported in the gastric cancer drug-resistant cell line SGC7901/ADR ? Our previous study showed that the expression of ?-catenin in Wnt/?-catenin pathway in SGC7901/ADR was significantly higher than in SGC7901 cell line.?-catenin is involved in the process of gastric cancer MDR.In S100A6 down regulated cell line(?-catenin is also showed low expression),it is a positive correlation.Both survivin and cyclin D1 are controversial factors in the downstream of Wnt/?-catenin,and their role in multidrug resistance of tumor cells is not clear.In this study,we used Western Blot,immunocytochemistry,RT-PCR and CCK8 to detect the expression of survivin and cyclin D1 in gastric cancer cell line SGC7901 and SGC7901/ADR cells.We used human gastric cancer cell line SGC7901 and its drug resistant cell SGC7901/ADR and ?-catenin cells lower,S100A6 plasmid,which was constructed by the previous group,to reduce the expression of sh RNA-1 and sh RNA-3 plasmid.By observing the expression of survivin and cyclin D1 in different cell lines,we explored the relationship among Wnt/?-catenin,survivin and cyclin D1.It provides a theoretical basis for the clinical treatment of tumor and the multi drug resistance of tumor.Objective: To investigate the relationship between survivin and cyclin D1 and multidrug resistance of gastric cancer cell line SGC7901/ADR.To explore the relationship between survivin,cyclin D1 and Wnt/ beta-catenin about gastric cancer MDR.Method:1.Immunocytochemistry,Western blotting and RT-PCR were used to detect differential expression about survivin and cyclin D1 in SGC7901 cells and the adriamycin resistant SGC7901/ADR cells.2.We used immunocytochemistry,Western Blotting Technology and RT-PCR technology to detect survivin and cyclin D1 differential expressions in S100A6 down to ?-catenin low expression of SGC7901 cells:sh RNA-1 stably transfected plasmid cells,sh RNA-3 plasmid stably transfected cells.3.CCK-8 cytotoxicity assay was used to detect the effects of adriamycin(ADR)on the half inhibitory concentration(IC50)and resistance index(RI)of the five kinds of cells.Result:1.Survivin:(1)immunocytochemistry: survivin in SGC7901 cytoplasm stained brown,was strongly positive(+++);SGC7901/ADR cytoplasm and some nuclei showed brown staining was strongly positive(+++);SGC7901,NC plasmid brown staining was positive(+++),and sh RNA-1?sh RNA-3 plasmid stably transfected cells cytoplasm staining with brown staining was positive(++).(2)Western blotting showed that survivin expression in SGC7901/ADR cells was higher than that of SGC7901 cells.The difference was obvious(p<0.05);The average expression of SGC7901 cells was significantly higher than that of S100A6 down regulated sh RNA-1 and sh RNA-3 plasmid stable cells with low expression of ?-catenin,and the difference was significant,(p<0.05).(3)the results of RT-PCR showed that the expression of SGC7901/ADR cells was higher than that of SGC7901 cells,and the difference was significant(p<0.05).The average m RNA expression of survivin in SGC7901 cells was significantly higher than the S100A6 was downregulated by strain sh RNA-1 and sh RNA-3 plasmids stably transfected cells to ?-catenin reduced,there was significant difference,p<0.05.2,Cyclin D1 results showed that(1)immunocytochemical: cyclin D1 in SGC7901 the nucleus brown,positive(++),SGC7901/ADR nucleus showed brown staining was strongly positive(+++);SGC7901 and NC plasmids were stably transfected cell cytoplasm was yellew-brown,positive(++)and sh RNA-1;the sh RNA-3 plasmid stable transfection cell cytoplasm staining,staining was negative(-).(2)Western blotting showed that the average expression of SGC7901/ADR cells was significantly higher than that of SGC7901 cells,there were significant differences(p< 0.05);the average expression was higher than that of S100A6,sh RNA-1 and sh RNA-3 strains by plasmid stable transfection of SGC7901 cells,there were significant differences,p<0.05.(3)RT-PCR :cyclin D1's m RNA of SGC7901/ADR is higher than SGC7901,there was significantly higher(p<0.05);m RNA SGC7901 cells average expression was higher than that of S100A6 down regulated sh RNA-1 and sh RNA-3 strains by plasmid stable transfection cells p<0.05,there was significant difference.Conclusion 1.Survivin and cyclin D1 may be related to the drug resistance of gastric cancer cell line SGC7901/ADR,survivin and cyclin D1 were highly expressed,and the drug resistance increased.2.Survivin and cyclin D1 may be downstream of the Wnt/?-catenin signaling pathway,involved in multidrug resistance of gastric cancer.3.High expression of beta-catenin,increased expression of Survivin and cyclin D1,low expression of beta-catenin,decreased expression of Survivin and cyclin D1,and the expression of beta-catenin and Survivin,beta-catenin and cyclin D1 were consistent... |
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