[Reversing effect of brassinolide on multidrug resistance of-CCRF-VCR1000 cells and a preliminary investigation on its mechanisms]

Aim: To investigate the effect of Brassinolide, a plant growth modulator, on multidrug resistance (MDR) of human T lymphoblastoid cell line CCRF-VCR 1000 which was obtained by progressively addition of vincristine (VCR) to sensitive CCRF-CEM cells, and to explore preliminarily the mechanism of reversing action.

Methods: MTT method was used to detect the resistant factor of resistant cell line and the reversing fold after addition of Brassinolide. The intracellular accumulation of rhodamine 123, a Fluorescent Dye transported by P-glycoprotein was detected by flow cytometry, the catalytic activity of Topoisomerase II was assessed by Sulliven method to find the effect of Brassinolide on resistance. The protein expression of p53 was measured using Western blotting in the sensitive cells and resistant cells to explore the effect of Brassinolide.

Results: The resistant factors of CCRF-VCR cells on adriamycin, VP-16 and VCR are respectively as 153.1, 55.9 and 8123.1 folds comparing to the sensitive cell line CCRF-CEM. After treatment of Brassinolide under the concentration of 0.001 - 10.0 microg x mL(-1), the resistance of CCRF-VCR was reversed partly with the reversing folds respectively as 4.4 - 11.6. The intracellular accumulation of rhodamine 123 was significantly reduced in the resistant cells. After treatment of Brassinolide, the accumulation increased, the level of Fluorescent Dye was situated between resistant cells and sensitive cells. No alteration of the catalytic activity of Topoisomerase II was found among three groups. The level of protein expression of p53 in resistant cells was higher than that of sensitive cells. After Brassinolide treatment, the expression of p53 in CCRF-VCR cells restored to the level of sensitive cells.

Conclusion: Brassinolide could effectively reverse the resistance of CCRF-VCR cells by inhibiting the effusion of drug transported by P-glucoprotein. To down regulate the abnormal expression of p53 maybe one of the mechanisms of reversing MDR for Brassinolide.