Synergistic Effect Of Bortezomib Combined With Pirarubicin On T Cell Lymphoma Cell Line (Hut-78)

T cell non-Hodgkin's lymphoma has a high incidence in China,approximately composes more than 30%in all non-Hodgkin's lymphoma.At present,neither domestic nor abroad has ideal treatment,the efficiency of conventional therapy of the CHOP regimen can be recently,but long-term effect and the prognosis of patients are very poor.Thus it is urgently to find effective anti-cancer drugs or combined regimen,which can improve the effect and the prognosis of patients.Bortezomib was the first proteosome inhibitor on the clinical research,which has been used for multiple myeloma and some B cell non-Hodgkin's lymphoma,the anti-tumor effects of T-cell lymphoma is not very clear.This experiment will investigate anti-tumor effects of bortezomib on T cell non-Hodgkin's lymphoma(Hut-78 cell line) in vitro,and observe whether the combination of bortezomib and pirarubicin have synergistic or superimposed effect on Hut-78 cells.Analysis the effect of proliferation and apoptosis induced by sequential administration,and analysis its possible mechanism preliminary.Then can provide a theoretical basis to help us choose the best treatment regimen.Methods:1 we used MTT assay to examine the effects of bortezomib on the proliferation of Hut-78 cell line.2 we used MTT assay to examine the effects of bortezomib combine with pirarubicin on the proliferation of Hut-78 cell line.3 PI staining assay were used to detect cell cycle block effect of Hut-78 induced by bortezomib,pirarubicin monotherapy and sequential administration.4 we used Annexin-V assay to detect apoptosis of Hut-78 cell induced by bortezomib,pirarubicin monotherapy and sequential administration.Results:1 The inhibition of bortezomib and pirarubicin on the proliferation of Hut-78 cells. MTT assay results showed that different concentrations of bortezomib or pirarubicin monotherapy can inhibit tumor cell proliferation when co-cultured with Hut-78 cells after 24h and 48h.The pirarubicin inhibition rate and drug concentration was positively correlated(P<0.01),in a dose-dependent manner;and 48h inhibition rate was significantly higher than 24h(P<0.01),in a time-dependent manner.2 The inhibition of bortezomib combined with pirarubicin in different order of administration on the proliferation of Hut-78 cell line.Hut-78 cells were treated 48h by three means of administrtion:first pirarubicin then bortezomib(THP→BTZ),Bortezomib and pirarubicin the same time(BTZ+THP), first bortezomib then pirarubicin(BTZ→THP).The results showed both BTZ+THP and BTZ→THP group had synergistic effect.BTZ→THP group was more significant, with the increase of drug concentration,the effect of tumor inhibition rate(fa) gradually increased,the combine index(CI) value decreased;BTZ + THP group with the concentration increased,fa gradually increased,CI increased.THP→BTZ group had synergistic effect in a certain range.So,BTZ→THP group had the best synergistic effect.3 Bortezomib monotherapy blocked cells at G2/M phase in a dose-dependent form,while pirarubicin monotherapy had no significant effect on cell cycle. Sequential administer pirarubicin can increase BTZ-induced G2/M detention significantly.4 Both bortezomib and pirarubicin monotherapy can inducd cells apoptosis in a dose-dependent form.The apoptosis rate of BTZ→THP group was significantly higher than bortezomib or pirarubicin monotherapy.Conclusion:Bortezomib combine with pirarubicin can inhibit proliferation of Hut-78 cell significantly,and induce apoptosis of cells.Sequential administration can induce higher synergistic cytotoxic effects.This study shows that bortezomib sequential pirarubicin provides a experimental basis for T cell non-Hodgkin's lymphoma.