Up-Regulation of Bax/BCL-2 Ratio by 2-Methoxyestradiol Induces Apoptosis in Lymphoma Raji Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of 2-methoxyestradiol (2-ME2) to lymphoma Raji cells and its mechanism.@*METHODS@#Different concentrations of 2-ME2 were used to treat lymphoma Raji cells. CCK8 method was used to detect the effect of 2-ME2 to proliferation of Raji cells. Flow cytometry FITC/PI double labeling method was used to detect early apoptosis of the cells. Western blotting was used to detect the effect of 2-ME2 to the expression of BCL-2, Bax, Caspase-3 and C-myc proteins in Raji cells.@*RESULTS@#2-ME2 significantly inhibited the proliferation of Raji cells. The inhibition rate increased with the increasing of drug concentration, and increased significantly with the prolongation of drug treatment time (r=0.9215). Flow cytometry FITC/PI double staining showed that the apoptotic rate of 2.5 μmol/L 2-ME2 treatment group was (33.79±1.63) %, while the apoptosis rate of the 48 h group was (51.90±2.72) %, and that of the control group was (7.08±0.36) %. After treated with 2.5 μmol/L 2-ME2 for 12 h, the expression of Bax protein was up-regulated, BCL-2 protein was down-regulated, caspase-3 protein expression was up-regulated, and C-myc protein expression was down-regulated, all of them showed a time-dependent relationship.@*CONCLUSION@#2-ME2 shows obvious inhibitory effect on lymphoma Raji cells in a dose- and time-dependent manner. Its mechanism of treatment on lymphoma Raji cells may be related to up-regulation of Bax/BCL-2 ratio and activation of Caspase-3 to induce apoptosis in cancer cells. Down-regulation of C-myc protein expression also participates in the apoptotic process.

Effect of Chlorambucil on the Apoptotic Signaling Pathway in Lymphoma Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study whether chlorambucil has apoptotic effect on the B cell lymphoma A20 cells and its exact mechanisms in apoptotic signaling pathway.</p><p><b>METHODS</b>The experimental cells were treated with 20 μmol/L chlorambucil, the control cells were treated with PBS. Annexin V-FITC Cell Apoptosis Detection Kit was used to examine cell apoptosis. Western blot was used to detect the expressions of active caspase-3, Survivin, NF-B and pAKT. Real-time fluorescent quantitative PCR was performed to examine the mRNA expression of Survivin.</p><p><b>RESULTS</b>Compared with the control group, the proportion of FITC/PI apoptotic cells and the expression of active caspase-3 (t=7.384, P=0.000) in the chlorambucil treatment group was significantly elevated. However, the expression of Survivin mRNA (t=4.384, P=0.000), protein expressions of survivin (t=12.360, P=0.000), NF-B (t=5.462, P=0.000) and pAKT (t=7.183, P=0.000) in the chlorambucil-treated group all significantly decreased.</p><p><b>CONCLUSION</b>The chlorambucil can induce the apoptosis of lymphoma cells, its mechanism may related with inhibition of PI3K/AKT signaling pathway, and expression of NF-B and survivin.</p>

Clinical Characteristics and Prognosis of patients with Variant Philadelphia Chromosome Positive Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the clinical characteristics and prognosis of patients with variant Ph chromosome-positive leukemia.</p><p><b>METHODS</b>The defection of morphology, cytogenetics, immunology and molecular biology was performed in 4 cares of variant Ph chromosome-positive leukemia, and the therepeuitics outcome of 4 patients was evaluated.</p><p><b>RESULTS</b>Among 4 cases of variant Ph+ leukemia, 3 cases were patients with CML, including 1 case in chronic phase and 2 cases in accelerated phase; and 1 cases was patient with adult B acute lymphoblasric leukemia(B-ALL).The defecfion of cytogenetics in 4 cases showed that 2 cases of CML displayed t(9; 22; 14) abnormality, 1 case of CML displayed t(5; 9; 22) abnormality, moreover, the BCR/ABL fution gane in 3 cases of CML all was e14a2 type, 1 cases of adult B-ALL disylayed t(9; 22; 17) abnormatlity, BCR/ABL fution gene of this case was e13a3 type, 4 patients all received treatment wire chemotherapeptic regimen contaiming methanesulfanate imatinib. As a result, 1 cases of adult B-ALL with e13a3 type BCR/ABL fusion gene positive relapsed after molecular biology remission for 4 months and died in the 10th month; and yet 3 cases of CML are still in molecular biology remission, the disease-free survival time of these 3 cases was 10, 19 and 27 months respectively.</p><p><b>CONCLUSION</b>The patients with variant Ph chromosome-positive leukemia will response to the first generation tyrosine kinase inhibitors, but the prognosis of patients with e13a3 type of BCR/ABL fusion gene remains to be further explored.</p>

Clinical Analysis of EB Virus Infection Complicated with Hemophagocytic Syndrome and Hodgkin's Lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical characteristics and outcome of parhents with EBV infection conbined with hemophagocytic syndrome and Hodgkin's lymphoma.</p><p><b>METHODS</b>The morphotogy of bone marrow cells was observed by bone marrow smear and light microscopy, the pathologic changes of bone marrow ware analyzed by bone marrow biopsy and immunohistochemistry methord, the pathologic changes of lymphonudes ware detected by immunohistochemical methord, the paticnts were treated with ABVD （epirubicin, bleomycin, vincristine and dacarbazine） chemotherapeutic regimen.</p><p><b>RESULTS</b>Fever complicatid with pancytopenia, obvious increase of ferritin and sCD25, hypofibrinogenemia, hemophogocytic phenomen of bone marrow, increase of EBV-DNA copy number ware observed, which all accorded with the criteria EBV righted hemophagocytic syndrome. The curative efficacy of amtiinfective treatmatnt was poor, After treatment with HLH-2004 regimen, the fever symptome and the laboratory indicaters such as whole blood cells, ferritin and fibrinogen all were recovered to normal levels. Left mandibular lymphadenctasis was confirmed as Hodgkin's lymphoma （mixed cell type） by pathological examination. The patient achieved complete molecular remission after 1 course chemotherapy with ABVD regimen. The level of EBV-DNA copy number were also decreased. As the reshlt, the patient's hemophagocytic syndrome had bean effectively controlled, and the Hodgkin's lymphoma is still in complete remission.</p><p><b>CONCLUSION</b>Epstein-Barr virus-ratated hemophagocytic syndrome and Hodgkin's lymphoma are rare, and their long-term prognosis needs to be further explored.</p>

Curative Effect of Decitabine Combined with IAG Regimen for Senile Patients with Myelodysplastic Syndrome (MDS) Transformed Acute Myeloid Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the curative effect and safety of decitabine combined with IAG regimen for treating senile MDS-transformed AML patients.</p><p><b>METHODS</b>Two cases of senile MDS-transformed AML were treated with decitabine combined with IAG regimen (decitabine 25 mg/d,qd,ivgtt,d1-5,Idarubicin 10 mg/d,qd,ivgtt,d6,Ara-C 10 mg/m,q12h, sc,d 6-19,G-CSF 300 µg,qd,ih,d6-19). The efficacy and adverse reactions were observed in these cases.</p><p><b>RESULTS</b>1 case for 2 courses and 1 case for 1 course obtained complete remission(CR). The myelosuppression and infections due to neutropenia were the most frequent adverse effects, the severe nonhematologic toxicity, such as liver and kidney and gastrointestinal reactions, were not observed in these patients.</p><p><b>CONCLUSION</b>Decitabine combined with IAG regimen is an effective for treating senile MDS-transformed AML patients.</p>

Change of TK1 Expression Level in NHL Patients before and after Chemotherapy and Its Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the change of TK1 expression level in patients with NHL and its clinical significance.</p><p><b>METHODS</b>A total of 108 NHL patients from July 2013 to January 2016 were chosen, all patients were treated with chemotherapy. The TK1 expression level and TK1 variation rate were compared among CR/PR/PD/DR patients. All patients were followed up, and the relation of TK1 level with OS/PFS was analyzed.</p><p><b>RESULTS</b>After treatment, the TK1 expression level of CR/PR/SD patients decreased significantly (P<0.05). Before treatment, the TK1 expression level of CR patients was 1.49±0.34,that after treatment was 0.45±0.17,the variation rate was (68.12±5.41)%; those of PR patients were respectively 2.89±0.58, 1.43±0.29 and (50.27±4.82)%; those of PD patients were respectively 3.98±0.78, 3.71±0.85 and (5.04±0.31)%; and those of SD patients were respectively 3.49±0.92, 2.45±0.57 and (28.65±3.97)%, the difference had statistic significance(P<0.05). After treatment, TK1 level related significantly with OS and PFS (P<0.05).</p><p><b>CONCLUSION</b>TK1 expression level has suggested implications for the evaluation of tumor loading, and treatment effect as well as prognosis of patients with NHL.</p>

Action Mechanism of Chlorambucil against Mantle Cell Lymphoma Cell Line Jeko-1 / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the action mechanism of chlorambucil against mantle cell lymphoma cell line Jeko-1.</p><p><b>METHODS</b>The effect of chlorambucil on Jeko-1 cell proliferation was measured by MTT method. The effect of chlorambucil on the apoptosis of Jeko-1 cell was detected by Hoechst staining and Annexin V-FITC dual staining. The activation of PI3K/AKT signaling pathway and the expression of BAX, BCL-2, procaspase 3, procaspase 8 and procaspase 9 were detected by Western blot.</p><p><b>RESULTS</b>0, 5, 10, 20 µmol/L chlorambucil could inhibit Jeko-1 cell proliferation at 24, 48, 72 h in a time- and dose-dependent manner. Chlorambucil of 0, 5, 10, 20 µmol/L increased the apoptotic rate of Jeko-1 cells, upregulated the expression of BAX, procaspase 3, procaspase 8, procaspase 9 and PI3K, increased the phosphorylation of AKT and down-regulated the expression of BCL-2.</p><p><b>CONCLUSION</b>The chlorambucil can induce the apoptosis of mantle cell lymphoma Jeko-1 cells via blocking PI3K/AKT signaling pathway.</p>

2-methoxyestradiol induced apoptosis and expression of p53 gene in human acute T lymphoblastic leukemia cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of 2-methoxyestradiol (2-ME2) on apoptosis of human acute T lymphoblastic leukemia cells, and its underlying mechanism.</p><p><b>METHODS</b>The growth inhibition of CEM cells was detected by MTT assay; apoptotic cells were detected by DNA laddering analysis; the expressions of P53 mRNA and protein were detected by RT-PCR and Western blot respectively.</p><p><b>RESULTS</b>2-ME2 remarkably inhibited the CEM cell growth and the 50% growth inhibitory concentration (IC50) at 48 h was 2 µmol/L. The DNA ladder could be detected in CEM cells after treating with 2 µmol/L 2-ME2 for 24, 48 and 72 hours; after treating with 2 µmol/L 2-ME2 for 24, 48 and 72 hours, a time-dependent reduction of P53 mRNA and protein expressions was found in CEM cells.</p><p><b>CONCLUSION</b>The anti-leukemia effect of 2-ME2 is completed through the induction of cell apoptosis. Down-regulation of P53 gene expression may be an underlying mechanism.</p>

Effect of reduced glutathione as anti-leukemic immune adjuvant / 中国实验血液学杂志

To investigate the reversal effect of reduced glutathione (GSH) on suppression of NK cells by reactive oxygen metabolites (ROM) in K562 cells, interleukin-2 (IL-2) or mononuclear cell (Mo) was added in cultured cell line of K562 cells and NK cells, the yield of ROM and K562 cell suppression rate were observed. Then the histamine dihydrochloride (DHT) or GSH was added in the mixed cultured cell lines, the ROM production and K562 cell suppression rate were observed. The results showed that the ROM yield increased from 33.17 +/- 5.08 U/L to 223.59 +/- 9.41 U/L by IL-2, and K562 cell suppression rate increased from 65.56% to 85.89% by IL-2 (P < 0.01). The ROM yields were 389.79 +/- 43.83 U/ml, 456.74 +/- 42.77 U/ml and 601.42 +/- 21.92 U/ml respectively, and K562 cell suppression rates were 82.36%, 81.36% and 48.09% respectively, when Mo was added in the mixed cultured cell lines under ratios of E/Mo being 10/2, 10/5 and 10/10. When E/Mo was 10/2, DHT or GSH was added in the mixed cultured cell line ROM yield decreased from 389.79 +/- 3.83 U/L to 50.21 +/- 2.4 U/L or -3.58 +/- 9.49 U/L (P < 0.05) respectively. With increase of concentration of DHT or GSH, the ROM yield in the mixed cultured cell line decreased (P < 0.05), the K562 cell suppression rate increased from 82.53% to 94.64% or 96.39% (P < 0.05), the more ROM yield, the less K562 suppression rate (P < 0.05). When E/Mo is 10/5 or 10/10, the ROM yield decreased by the high concentration of DHT or GSH (P < 0.05), but the K562 cell suppression rate not increased by every concentration of DHT or GSH. GSH was as effective as DHT in the reversing ROM and increasing K562 cell suppression rate. It is concluded that GSH may reverse ROM and increase K562 cell suppression rate, and GSH is as effective as DHT, but GSH has less side-effect than DHT. Therefore, GSH would be better antileukemia immune adjuvant.