Comparison Of The Antitumor Effects Of Cucurbitacin D And Cucurbitacin I On T-cell Lymphoma Cells

T-cell lymphoma is a highly aggressive disease with a poor prognosis. According to the World Health Organization(WHO) classification of lymphoid neoplasms in 2008, there are more than 10 kinds of subgroup related with T cells in group of mature T-cell and NK-cell neoplasms. NK/T-cell lymphomas occur worldwide, with a strong geographic predilection for Asian populations from China, Japan, Korea, and Southeast America population. T-cell lymphoma is more aggressive, and the prognosis is poorer compared with B-cell lymphoma. Unfortunately, there is no standard treatment strategy for T-cell lymphomas now. Therefore, development of new drugs for this aggressive disease is urgently needed.Cucurbitacins belong to a group of triterpenoids isolated from plant families such as Cucurbitaceae and Cruciferae, which have been shown to have antipyretic, analgesic, anti-inflammatory, antimicrobial, and antitumor activities. Cucurbitacins are divided into 17 categories, including cucurbitacin A to T. Cucurbitacin B, D, E and I have been reported for their anticancer activities. Interestingly, the chemical structures of Cucurbitacin D and I show strong similarity.Considering the similarities and differences of chemical structure of Cucurbitacin D and I, we investigated the relationship between structure and their anti-tumor effects on T-cell Lymphoma, which will help for development of new drugs for T-cell Lymphoma. The main research contents were shown as follows: PartⅠ Comparison of the Antitumor Effects of Cucurbitacin D and Cucurbitacin I on Mouse T-cell lymphoma Cell LineEL4Objective: The antitumor effects of cucurbitacin D and cucurbitacin I on mouse T-cell lymphoma cell line EL4 were compared, to clarify the relationship between their structure and anti-tumor effect, and provide a new approach for treating T-cell lymphomas.Methods:1 EL4 cells were cultured in DMEM medium, and passaged every 24 to 48 hours. The cells were used for experiments during logarithmic growth phase.2 The effectiveness of cucurbitacin D and I on the proliferation of mouse T-cell lymphoma cell line EL4 were evaluated using a Cell Titer-Glo luminescent cell viability assay system.3 The effects and mechanism of cucurbitacin D and I on apoptosis of mouse T-cell lymphoma cell line EL4 were compared by western blot analysis for the expression of pro-apoptotic proteins-- PARP and Caspase-3.4 The level of NF-κB p65 and phospho- NF-κB p65 were detected by western blot analysis for checking the anticancer mechanisms of cucurbitacin D and I on EL4 cell line.Results:1 Results from Cell Titer-Glo luminescent cell viability assay indicated that cucurbitacin D and I inhibited cell proliferation of T-cell lymphoma cell line EL4. Cucurbitacin I was more effective than cucurbitacin D on EL4 cell line(P<0.05).2 Data from western blot analysis showed that both cucurbitacin D and I suppressed the expression of apoptosis-related proteins cleaved-caspase-3 and cleaved-PARP. Cucurbitacin I showed stronger effect than cucurbitacin D(P<0.05).3 Cucurbitacin D and cucurbitacin I inhibited activation of NF-κB p65, and cucurbitacin D had stronger inhibition effect on NF-κB p65 phosphorylation than cucurbitacin I(P<0.05).Conclusions:1 Cucurbitacin D and Cucurbitacin I inhibit proliferation of EL4 cells by suppressing of NF-κB p65 phosphorylation, caspase-3 and PARP cleavage, induceing apoptosis.2 The inhibition effect of Cucurbitacin I on EL4 cells was higher than Cucurbitacin D. PartⅡ Comparison of the Antitumor Effects of Cucurbitacin D and Cucurbitacin I on Human T-cell Lymphoma CellLines and Peripheral Blood Lymphocytes from T-cellLymphoma Patients and Healthy Donors.Objective: The antitumor effects of cucurbitacin D and cucurbitacin I on human T-cell lymphoma cell lines and peripheral blood lymphocytes from T-cell lymphoma patients and healthy donors were compared, to evaluate deeply cucurbitacin D and cucurbitacin I for T-cell lymphoma treatment.Methods:1 Human T-cell lymphoma cell lines Hu T78, Hu T102, MT-2, MT-4 were cultured in DMEM medium, and passaged every 24 to 48 hours. The cells were used for experiments during logarithmic growth phase.2 The effects of cucurbitacin D and I on the proliferation of Human T-cell lymphoma cell lines Hu T78, Hu T102, MT-2, MT-4 were examined using a Cell Titer-Glo luminescent cell viability assay system.3 Peripheral blood lymphocytes(PBLs) were prepared from 12~20m L EDTA-blood from human T-cell lymphoma patients and healthy donors using Lympholyte.4 The effects of cucurbitacin D and I on the proliferation of peripheral blood lymphocytes(PBLs) from T-cell lymphoma patients and healthy donors were compared using a Cell Titer-Glo luminescent cell viability assay system.Results:1 Cucurbitacin D and I inhibited cell proliferation of Human T-cell lymphoma cell lines Hu T78, Hu T102, MT-2, MT-4. Cucurbitacin I showed stronger effect than cucurbitacin D(P<0.05).2 Cucurbitacin D and I inhibited cell proliferation of Peripheral blood lymphocytes(PBLs) from human T-cell lymphoma patients(P<0.05), but not healthy donors(P>0.05).Conclusions:1 Cucurbitacin D and I inhibited cell proliferation of Human T-cell lymphoma cell lines Hu T78, Hu T102, MT-2, MT-4 and peripheral blood lymphocytes(PBLs) from human T-cell lymphoma patients. Cucurbitacin I showed stronger effect than cucurbitacin D.2 Cucurbitacin D and I inhibited cell proliferation of peripheral blood lymphocytes(PBLs) from human T-cell lymphoma patients, but not PBLs from healthy donors.