| Objective: The purpose of this study is to establish a oxaliplatin resistant CT-26-R and SW-480-R cell line,so as to provide a theoretical basis for preliminary investigation of the mechanism of drug resistance.Methods: to establish oxaliplatin resistant cell line CT-26-R using low concentration long time contact resistance method,two kinds of cells were detected by CCK-8 method in SW-480-R.two cell line was successfully established,the growth curves of the two kinds of cells,finally by Western blotting in two TNF-a resistant cell detection in detecting the content of P-pg.Results: After six months of time eventually won the grow well,tolerated(including 250.38 g/mL OXP resistant cells CT-26-103.3 ug/R cell lines and tolerance mlOXP SW-480-R cell line.Two kinds of cell growth curve as shown in figure,the cell growth of CT-26-R than CT-26 cells obviously slow,CT-26-R cells of TD is 52.8 h,CT-33.6 h 26 cells of TD,CT-26 TD is computed tomography(CT)-26-R cells TD's 1.57 times.SW-480-R cell lines of TD is 50.4 h,SW-480 cells of TD is 48.1 h,SW-480 cells of TD is SW-480-R cells TD's 1.25 times.Both drug resistant cell lines showed the phenomenon of slow growth.CCK 8 method to detect cellular activity can be found that CT-26-R,SW-480-R cell for tolerance of oxaliplatin into was obviously higher than that of CT-26,the SW-480 cell line,two kinds of drug-resistant cell line successfully established,and approved by the cryopreserved can successfully raised,can provide stable cell material for future experiments.At the same time through protein imprinting method to detect the sw-480 and sw-480-R E-cadherin protein content in the two cell lines,visible E-cadherin protein in drug resistance of oxaliplatin into sw-480-R cells significantly less resistant sw-480 cells,the difference was statistically significant(p < 0.05),and p-pg protein and TNF-a factor in the resistance of sw-480-R is a sw-480 cells increased significantly,the difference was statistically significant(p < 0.05);And similar results also appeared on CT-26 and computed tomography(CT)-26-R cell lines,resistance of oxaliplatin into CT-26-E-R cells cadherin protein less resistant CT-26 cells decreased obviously,the difference was statistically significant(p < 0.05),and p-pg protein and TNF-a factor in the resistance of CT-26-R cells is a CT-26 cells increased significantly,the difference was statistically significant(p < 0.05).By protein imprinting results can be seen,two not drug resistance cell line between SW-480 and CT-26,CT-26 of TNF-a and P-pg protein and E-cadherin protein were higher than that of SW-480,the drug resistance of SW-480-R and computed tomography(CT)-26-R,TNF-a and P-pg protein did not see obvious difference,and E-cadherin protein content in SW-480-R cells was higher than that of CT-26-R.Conclusion: This experiment successfully established oxaliplatin into drug-resistant CT-26-R and SW-480-R cell lines,including two kinds of drug resistance cell line of TNF-a and P-pg content increased to some extent,E-cadherin protein content decreased,so the EMT process enhancement,P-pg protein,and increased levels of TNF-a tumor cell resistance there is a certain correlation.The two cell lines established in this experiment could eventually be used in other similar studies. |
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