The Effect Of Emodin Derivatives On T Lymphocytic Leukemia Molt-4and Resistant Molt-4/ADR Cells

[Objective]（1）To study the influence of Emodin derivatives E19and E26on proliferation and apoptosis in human T lymphocytic leukemia Molt-4cell lines and resistant Molt-4/ADR cell lines.（2）To observe the changes of the cell proliferation, apoptosis, drug resistance and signaling pathway-related proteins, after Emodin derivative E19and E26treatment on Molt-4and Molt-4/ADR cell lines.[Methods]（1）MTT method and cell colony formation assay were chosen for cell proliferation test. MTT method was used to observe the change of cell growth curves and the number of cell clones was counted based on the drug acting on the cells.（2）DAPI staining and DNA ladder test were used for the test of cell apoptosis. The changes of stained cellular morphology under the fluorescence microscope were observed. The DNA ladder images were observed based on the drug acting on the cells.（3）Western blot was used to observe the expression changes of cell apoptosis related proteins, such as bcl-2, procaspase-3, and PARP, etc., drug-resistant related proteins, such as P-gp, and PI3K/AKT, MAPK signaling pathway-related proteins based on the drug acting on the cells.[Results]（1）Using MTT method, it could be observed that emodin derivatives E19and E26of different concentration on Molt-4and Molt-4/ADR cells or treatment at different time significantly inhibited the proliferation of cell lines, showing dependent on concentration and time. For Molt-4and Molt-4/ADR cells, the median inhibitory concentration of E19within48hours (IC50) was1.199±0.204μmol/L and15.60±1.082μmol/L, respectively, and E26’s IC50was1.390±0.152μmol/L and8.564±0.546μmol/L.（2）From cell colony forming units, it could be observed that emodin derivatives E19and E26with the increase of drug concentration on Molt-4and Molt-4/ADR cells, the number of cell colony formation gradually decreases, so emodin derivatives have a significant inhibitory effect.（3）From DAPI staining method, typical morphologic change of apoptosis cells under the fluorescence microscope could be found, when emodin derivatives of different concentrations acted on the cells.（4）From DNA ladder analysis, typical DNA degradation fragment could be found, when emodin derivatives of different concentrations acted on the cells.（5）Different concentrations E19、E26could down-regulate the protein expressions of bcl-2、procaspase-3、procaspase-9、PARP、c-myc、AKT、p-AKT、mTOR、p-mTOR、MAPK、p-MAPK、p-P70、p-4EBP1in Molt-4and Molt-4/ADR cells. Procaspase-8、4EBP1、P70did not change remarkably. Bcl-2，AKT、p-AKT、mTOR、p-mTOR、procaspase-9，procaspase-3、PARP、p-4EBP1、c-myc、p-MAPK、p-P70proteins were down-regulated after E19、E26treatment in Molt-4and Molt-4/ADR cells for different time. Procaspase-8、4EBP1、P70、MAPK remained no change. P-glycoprotein expression in Molt-4/ADR cell，remained no change.[Conclusion]（1）Emodin derivatives, both E19and E26can effectively inhibit the proliferation of T lymphocyte leukemia cell lines Molt-4and Molt-4/ADR, and induce them to apoptosis.（2）Caspase family proteins, bcl-2and c-myc participate in the proliferation inhibition and apoptosis induction by emodin derivatives E19and E26on Molt-4、Molt-4/ADR cells.（3）Emodin derivatives E19and E26may induce Molt-4and Molt-4/ADR cells to death by inhibit the signalling pathway of PI3K/AKT and MAPK.