curcumin and Precursor-T-Cell-Lymphoblastic-Leukemia-Lymphoma

Acute lymphoblastic leukemia (ALL) is the most common hematological malignancy affecting pediatric patients. ALL treatment regimens with cytostatics manifest substantial toxicity and have reached the maximum of well-tolerated doses. One potential approach for improving treatment efficiency could be supplementation of the current regimen with naturally occurring phytochemicals with anti-cancer properties. Nutraceuticals such as quercetin, curcumin, resveratrol, and genistein have been studied in anti-cancer therapy, but their application is limited by their low bioavailability. However, their cooperative activity could potentially increase their efficiency at low, bioavailable doses. We studied their cooperative effect on the viability of a human ALL MOLT-4 cell line in vitro at the concentration considered to be in the bioavailable range in vivo. To analyze their potential side effect on the viability of non-tumor cells, we evaluated their toxicity on a normal human foreskin fibroblast cell line (BJ). In both cell lines, we also measured specific indicators of cell death, changes in cell membrane permeability (CMP), and mitochondrial membrane potential (MMP). Even at a low bioavailable concentration, genistein and curcumin decreased MOLT-4 viability, and their combination had a significant interactive effect. While resveratrol and quercetin did not affect MOLT-4 viability, together they enhanced the effect of the genistein/curcumin mix, significantly inhibiting MOLT-4 population growth in vitro. Moreover, the analyzed phytochemicals and their combinations did not affect the BJ cell line. In both cell lines, they induced a decrease in MMP and correlating CMP changes, but in non-tumor cells, both metabolic activity and cell membrane continuity were restored in time. (4) Conclusions: The results indicate that the interactive activity of analyzed phytochemicals can induce an anti-cancer effect on ALL cells without a significant effect on non-tumor cells. It implies that the application of the combinations of phytochemicals an anti-cancer treatment supplement could be worth further investigation regardless of their low bioavailability.

Topics: Apoptosis; Cell Line; Cell Line, Tumor; Curcumin; Genistein; Humans; Phytochemicals; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Quercetin; Resveratrol

AbstractObjective: To explore the effect and mechanism of curcumin on human T-cell acute lymphoblastic leukemia (T-ALL) cell apoptosis induced by Mcl-1 small molecule inhibitors UMI-77.. T-ALL cell line Molt-4 was cultured, and the cells were treated with different concentrations of curcumin and Mcl-1 small molecule inhibitor UMI-77 for 24 h. The MTT method was used to detect the cell survival rate after different treatment; According to the results of curcumin and UMI-77, the experimental settings were divided into control group, curcumin group (20 μmol/L curcumin treated cells), UMI-77 group (15 μmol/L Mcl-1 small molecule inhibitor UMI-77 treated cells) and curcumin+ UMI-77 group (20 μmol/L curcumin and 15 μmol/L Mcl-1 small molecule inhibitor UMI-77 treated cells), MTT method was used to detect cell proliferation inhibition rate, Annexin V-FITC/PI double staining method and TUNEL staining were used to detect cell apoptosis, DCFH-DA probe was used to detect cell reactive oxygen species, JC-1 fluorescent probe was used to detect mitochondrial membrane potential, Western blot was used to detect the expression levels of apoptosis-related proteins and Notch1 signaling pathway-related proteins.. After the treatment of Molt-4 cells with different concentrations of curcumin and Mcl-1 small molecule inhibitor UMI-77, the cell survival rate was decreased (P<0.05); Compared with the control group, the cell proliferation inhibition rate of the curcumin group and the UMI-77 group were increased, the apoptosis rate of cell was increased, the level of ROS was increased, the protein expression of Bax, Caspase-3 and Caspase-9 in the cells were all increased, and the protein expression of Bcl-2 was reduced (P<0.05); Compared with the curcumin group or UMI-77 group, the cell proliferation inhibition rate and apoptosis rate of the curcumin+UMI-77 group were further increased, and the level of ROS was increased. At the same time, the protein expression of Bax, Caspase-3 and Caspase-9 in the cells were all increased, the protein expression of Bcl-2 was reduced (P<0.05); In addition, the mitochondrial membrane potential of the cells after curcumin treatment was decreased, and the proteins expression of Notch1 and HES1 were reduced (P<0.05).. Curcumin can enhance the apoptosis of T-ALL cells induced by Mcl-1 small molecule inhibitor UMI-77 by reducing the mitochondrial membrane potential, the mechanism may be related to the inhibition of Notch1 signaling pathway.. 姜黄素对UMI-77诱导急性T淋巴细胞白血病细胞凋亡的影响及其相关机制研究.. 探究姜黄素对Mcl-1小分子抑制剂UMI-77诱导的人急性T淋巴细胞白血病（T-ALL）细胞凋亡的影响及其相关机制.. 培养T-ALL细胞系Molt-4，用不同浓度的姜黄素和Mcl-1小分子抑制剂UMI-77分别处理细胞24 h，MTT法检测经不同浓度姜黄素和UMI-77分别处理后的细胞存活率；根据姜黄素和UMI-77的作用浓度，实验设置对照、姜黄素（20 μmol/L姜黄素处理细胞）、UMI-77组（15 μmol/L Mcl-1小分子抑制剂UMI-77处理细胞）和姜黄素+UMI-77（20 μmol/L姜黄素加15 μmol/L Mcl-1小分子抑制剂UMI-77处理细胞）共4组，MTT法检测细胞增殖抑制率，Annexin V-FITC/PI双染法和TUNEL染色检测细胞的凋亡情况，DCFH-DA探针检测细胞活性氧，JC-1荧光探针检测线粒体膜电位，Western blot检测细胞凋亡相关蛋白和Notch1信号通路相关蛋白的表达水平.. 经过不同浓度的姜黄素和Mcl-1小分子抑制剂UMI-77处理Molt-4细胞后，细胞存活率降低（P<0.05）；与对照组比较，姜黄素组和UMI-77组细胞增殖抑制率增加，细胞凋亡率升高，ROS水平提高，细胞内Bax、Caspase-3和Caspase-9蛋白表达均增加，Bcl-2蛋白表达降低（P<0.05）；与姜黄素组和UMI-77组比较，姜黄素+UMI-77组细胞增殖抑制率和凋亡率进一步增加，ROS水平提高，同时，细胞内Bax、Caspase-3和Caspase-9蛋白表达均增加，而Bcl-2蛋白表达降低（P<0.05）；此外，姜黄素处理后细胞的线粒体膜电位下降，Notch1和HES1蛋白表达均降低（P＜0.05）.. 姜黄素通过降低线粒体膜电位来增强Mcl-1小分子抑制剂UMI-77诱导的T-ALL细胞凋亡，其机制可能与抑制Notch1信号通路相关.

Topics: Apoptosis; Apoptosis Regulatory Proteins; bcl-2-Associated X Protein; Caspase 3; Caspase 9; Cell Line, Tumor; Curcumin; Humans; Myeloid Cell Leukemia Sequence 1 Protein; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Proto-Oncogene Proteins c-bcl-2; Reactive Oxygen Species; Sulfonamides; Thioglycolates

Multidrug resistance (MDR) and drug transporter P-glycoprotein (P-gp) represent major obstacles in cancer chemotherapy. We investigated 19 synthetic curcumin derivatives in drug-sensitive acute lymphoblastic CCRF-CEM leukemia cells and their multidrug-resistant P-gp-overexpressing subline, CEM/ADR5000.. Cytotoxicity was tested by resazurin assays. Doxorubicin uptake was assessed by flow cytometry. Binding modes of compounds to P-gp were analyzed by molecular docking. Chemical features responsible for bioactivity were studied by quantitative structure activity relationship (QSAR) analyses. A 7-descriptor QSAR model was correlated with doxorubicin uptake values, IC50 values and binding energies.. The compounds displayed IC50 values between 0.7±0.03 and 20.2±0.25μM. CEM/ADR5000 cells exhibited cross-resistance to 10 compounds, collateral sensitivity to three compounds and regular sensitivity to the remaining six curcumins. Molecular docking studies at the intra-channel transmembrane domain of human P-gp resulted in lowest binding energies ranging from -9.00±0.10 to -6.20±0.02kcal/mol and pKi values from 0.24±0.04 to 29.17±0.88μM. At the ATP-binding site of P-gp, lowest binding energies ranged from -9.78±0.17 to -6.79±0.01kcal/mol and pKi values from 0.07±0.02 to 0.03±0.03μM. CEM/ADR5000 cells accumulated approximately 4-fold less doxorubicin than CCRF-CEM cells. The control P-gp inhibitor, verapamil, partially increased doxorubicin uptake in CEM/ADR5000 cells. Six curcumins increased doxorubicin uptake in resistant cells or even exceeded uptake levels compared to sensitive one. QSAR yielded good activity prediction (R=0.797 and R=0.794 for training and test sets).. Selected derivatives may serve to guide future design of novel P-gp inhibitors and collateral sensitive drugs to combat MDR.

Topics: Antineoplastic Agents; ATP Binding Cassette Transporter, Subfamily B, Member 1; Cell Line, Tumor; Cell Survival; Curcumin; Doxorubicin; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Humans; Models, Molecular; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Quantitative Structure-Activity Relationship