Establishment of drug-resistant cell lines of human triple negative breast cancer and study on their characteristics / 国际检验医学杂志

ABSTRACT

Objective To establish docetaxel (Doc) resistant MDA‐MB‐231/Doc model and epirubicin (Epi) resistant MDA‐MB‐231/Epi mode from triple negative breast cancer cell line MDA‐MB‐231 and to explore their biological characteristics .Methods The MDA‐MB‐231/Doc and MDA‐MB‐231/Epi drug‐resistant cell lines were respectively established by gradually increasing Doc or Epi concentrations induction method in 12 months .The biological characteristics of the cell lines were compared by the cell mor‐phological observation ,MTT and flow cytometry ;the real‐time fluorescent quantitative PCR was used to detect multi‐drug resist‐ance gene (MDR1) mRNA expression;the expression of P glycoprotein(P‐gp) ,estrogen receptor (ER) ,progesterone receptor (PR) and human epidermal growth factor receptor 2 (Her‐2) was detected by Western Blot .Results After the 12‐month induction ,the established MDA‐MB‐231/Doc could grow stably in the medium containing 12 nmol/L Doc ,and MDA‐MB‐231/Epi could grow stably in the medium containing 800 nmol/L Epi;in the same drug concentration ,the growth proliferation rate of the drug resistant cell line was significantly higher than that of the parental generation cells ,their drug resistance indexes were 8 .32 times and 64 .93 rimes of parental generation sensitive cells ,moreover which showed the mutual cross drug resistant status .Compared to the parental generation cells ,the cells of stage G1 and G2 in two cell lines were increased and the cells of stage S were decreased ,with the prolon‐gation of drug withdrawal time ,the cell proliferation speed was accelerated .The expression level of MDR1 gene was increased in the two drug‐resistant cell lines ,which were 4 .05 times and 5 .96 times of parental generation cells respectively ,P‐gp protein expression was positive .Compared with the MCF‐7 cell line ,ER ,PR and Her2 expression in the MDA‐MB‐231 cell line was negative and typi‐cal triple negative breast cancer cell line .Conclusion The drug resistance cell lines of MDA‐MB‐231/Doc and MDA‐MB‐231/Epi are successfully established with stable growth and drug resistance .