Establishment Of Human Leukemia And Osteosarcoma Methotrexate-resistant Cell Line And Drug Resistance Detection

The treatment of many tumor especially for leukemia chemotherapy tend toachieve better res μlts for the first time. However, the frequent relapset, and each timerecurrence after treatment effect is reduced, es μlt in a failure treatment。Resistance oftumor cells to chemotherapy drug is the main obstacle to cancer treatment，thus，thestudy of tumor drug resistance and the solution is of great significance。MTX is Used in chemotherapy regimens for various diseases，such as acutelymphoblastic leukemia;Malignant lymphoma, non-hodgkin’s lymphoma and mycosisgran μloma, m μltiple bone marrow disease, osteosarcoma. The treatment with MTXalso associated with drug resistance. To reveal the mechanism of action of MTX andthe cause of treatment failure phenomena of resistance, the first step is theestablishment of MTX drug-resistant cell mode。A not mutate, not clear importforeign genetic resistance, and the resistance of which can be stably inherited to MTXdrug resistance cell line progeny，Can provide credible research object for the study。SEM，a children-B cell acute lymphoblastic leukemia cell line；U2-OS，one ofthe types of osteosarcoma cells。Based on the above two kinds of cells，MTXdrug-resistant cell models are set up respectively in the original continuous constantstim μlation and imp μlse stim μlating ways，we also have identified the resistantability of each cell line.This experiment through chromosome karyotype analysis, Cell total proteinexpression comparison, the control group is the original cell, we have identified that,against original cells, there is not mutation of the chromosomes from MTX-resistantcell; protein expression, not missing. MTX-resistant cells still possess the originalcells`corresponding life functions and characteristics. By flow cytometry, MTTmethod, we confirmed that cells of resistance groups showed an lower inhibition rateon the impact of different dose MTX than the original cell, prompt that drug-resistantcells reduced susceptibility to MTX. DHFR is the marker to MTX-resistance. Theexpression of DHFR from MTX-resisitante cells have tended to increase, Tipresistance groups of cells develop resistance.