Effects of Emodin Derivative on Cell Cycle, Apoptosis and NF-κB Pathway in Burkitt Lymphoma Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect and mechanism of a novel emodin derivative YX-18 on Burkitt lymphoma (BL) cells.@*METHODS@#MTT assay was used to detect the effect of YX-18 on the proliferation of BL cell lines CA46 and Raji. Annexin V-PE/7-AAD double staining assay was used for detecting the effect of YX-18 on the apoptosis of CA46 and Raji cells. PI/RNase staining was used to test the effect of YX-18 on CA46 and Raji cell cycle. JC-1 method was used to measure the changes of mitochondrial membrane potential after YX-18 treatment, and DAPI staining was used to detect the morphology of apoptotic cells. Western blot was used to analyze the distribution changes of NF-κB pathway protein (P65, P-P65, IκB, P-IκB) in the cytoplasm and cell nucleus, and also the expression changes of cyclin-related protein P21, CDK2, P-CDK2, Cycling D1, Cycling E1, and the apoptosis-related protein Caspase-3, Caspase-8, Caspase-9 and the proliferation-related protein C-MYC, BCL-2 by YX-18. Real-time fluorescence-quantitative PCR was used to evaluate the effects of YX-18 on mRNA levels of C-MYC and Ki-67 genes in CA46 and Raji cells, and EBNA-1 and EBER genes of EBV in Raji (EBV@*RESULTS@#Novel Emodin derivative YX-18 could effectively inhibit the proliferation of BL cell lines CA46 and Raji, showing a time-dependent effect (24, 48 and 72 h: r@*CONCLUSION@#The novel emodin derivative YX-18 can significantly inhibit the proliferation of Burkitt lymphoma cells, and induce the cell apoptosis and cycle arrest. The inhibitory effect of YX-18 on the proliferation of Burkitt lymphoma cells may be related with the effect of Caspase apoptosis pathway, the proliferation and apoptosis-related molecules, such as C-MYC and Ki-67, and also to the inhibition of NF-κB pathway.

Oral Mucosal Diffuse Large B-cell Lymphoma Caused by Long-term Oral Methotrexate:Report of One Case / 中国医学科学院学报

A case of primary oral mucosal diffuse large B-cell lymphoma(DLBCL)due to long-term use of methotrexate(MTX)for the treatment of rheumatoid arthritis(RA)was admitted to the Department of Hematology,Fujian Medical University Union Hospital.We analyzed and discussed the clinical features,diagnosis and treatment,and prognosis of specific malignant lymphoma induced by MTX in this RA patient.Our purpose is to improve the awareness and knowledge of other iatrogenic immunodeficiency-associated lymphoproliferative disorders of clinicians and pathologists.This study provides a new reference for the clinical diagnosis and treatment of MTX-associated DLBCL.

Inhibitory and Inducing Effects of Emodin Derivative E11 on Proliferation and Apoptosis of Multiple Myeloma Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To screen the most strong emodin derivative inhibiting the proliferation of multiple myeloma(MM) cells and to explore the inhibitory and inducing effects of emodin derivatives on proliferation and apoptosis of MM cell lines RPMI 8226 and U266.</p><p><b>METHODS</b>Sixteen emodin derivatives were designed and synthesized by using emodin as mother substance, then from which the emodin derivative E11 was screened for experiments. The MTT method and cell colony formation assay were used to observe the effect of E11 on proliferation of RPMI 8226 and U266, the fluorescent microscopy with DAFI staining was used to observed the morphological changes of MM cells treated with emodin dervative 11, the DNA fragmentation detection was used to detect the inducing apoptosis effect of E11 on RPMI 8226 and U266 cells treated with E11.</p><p><b>RESULTS</b>The MTT assay showed that after the RPMI 8226 cells were treated with 16 kinds of emodin derivatives for 48 hours, the 50% inhibition concentration（IC） of 14 emodin dervatives was between 0.83-34.68 µmol/L, except E10 and E15 because their IC could not be calculated. The IC of E11 for RPMI 8226 and U266 cells were 0.831±0.0453 µmol/L and 1.039±0.093 µmol/L, respectively. Cell colony formation assay showed that E11 could inhibit RPMI8226 and U266 cells' colony formation in dose-.and time- dependent manner (r=0.72). Cell apoptosis was observed in RPMI8226 and U266 cells by DAPI staining , and also by the detection of DNA fragmentation.</p><p><b>CONCLUSION</b>In the synthesis of 16 kinds of emodin derivatives, the inhibitory effect of E11 on prolife-ration of RPMI8226 cell was the strongest. E11 can remarkably inhibit proliferation and induce apoptosis of RPMI8226 and U266 cells.</p>

Effect of A Novel Emodin Derivative on Chronic Myelogenous Leukemia K562 Cells and Imatinib-resistant K562/G01 Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of a novel emodin derivative E19 on proliferation inhibition and apoptosis induction of human chronic myelogenous leukemia (CML) cell line K562 and imatinib-resistant CML cell line (K562/G01), and to clarify the involved mechanisms.</p><p><b>METHODS</b>MTT and colony formation test were used to detect the cell proliferation. Apoptotic induction effects were examined by DAPI staining method and DNA ladder assay. Western blot was performed to detect the changes of P210(Bcr-Abl) protein.</p><p><b>RESULTS</b>The emodin derivative E19 could efficiently inhibit proliferation and induce apoptosis in K562 and K562/G01 cells. IC50 of K562 cells and IC50 of K562/G01 cells were (1.20 ± 0.19) µmol/L and (1.22 ± 0.16) µmol/L, respectively. DNA fragmentation in K562 cells and K562/G01 cells confirmed that the E19 induced apoptosis in dose-dependent manner. Western blot showed that emodin derivative inhibited phosphorylation of P210 protein in K562 cells and K562/G01 cells and down-regulated the expression level of P210 in dose- and time-dependent manners.</p><p><b>CONCLUSION</b>The emodin derivative E19 can efficiently inhibit growth and induce apoptosis of K562 cells and K562/G01 cells, while the inhibition of phosphorylation of P210 protein and down-regulation of P210 protein expression may be involved in these processes.</p>

Inhibitory effect of pumpkin protein on expression of Notch signal in RPMI8226 myeloma cells / 中国实验血液学杂志

This study was aimed to explore the inhibitory effect of pumpkin protein (cucurmosin, CUS) on proliferation of RPMI8226 myeloma cells in vitro and its mechanism. Western blot was used to detect the expression level of Notch-1, Jagged-2, P-Akt and NF-KB in the myeloma cells treated by different concentrations of CUS. The results demonstrated that CUS could down-regulate the protein expression levels of Notch1, Jagged-2, P-Akt and NF-KB in the myeloma cells and with time-and concentration-dependent way, at the same time CUS could also decrease the expressions of BCL-2 and P-Akt. It is concluded that CUS can obviously inhibit the RPMI8226 cell proliferation in vitro, down-regulate the expression levels of Notch signal and its down-stream target genes. Therefore, Notch signaling pathway can be used as a new treatment target for multiple myeloma, and CUS may be become a potential new drug for regulating Notch signaling pathway.

Synthesis of emodin derivatives and their inhibiting effects on proliferation of leukemia cell lines / 中国实验血液学杂志

The aim of this study was to explore the inhibitory effect of newly synthesised emodin derivatives on the proliferation of leukemia cell lines and to select the most effective one from these emodin derivatives for further research. Emodin derivatives were synthesized by modifying the structure of emodin. MTT method was used to detect the proliferative inhibition in leukemia cell lines treated with emodin derivatives. The results showed that the half inhibitory concentration (IC50) for K562 cells treated with emodin derivatives E10-19 for 48 h was 0.84 - 12.01 µmol/L. E19 displayed the best anti-proliferative activity, while E16 and E17 did not show effects on K562 cells. Emodin derivative E19 was chosen for treating U937, NB4, Molt-4 and CA-46 cells, their IC50 for 48 h were 0.85, 0.9, 0.76, 0.8 µmol/L respectively. The IC50 of E19 for LQ2 cells was 3.60 µmol/L, and the IC50 range of E19 for normal human peripheral blood mononuclear cells at 48 h was 4.01 - 4.78 µmol/L. It is concluded that emodin derivative E19 can strongly inhibit the growth of leukemia cells and its inhibiting effect on proliferation of leukemia cells has a certain specificity. The specific mechanism of E19 anti-leukemia effect should be further studied.

Efficacy of CALLG2008 protocol in treatment of adult acute lymphoblastic leukemia: a single center analysis / 中国实验血液学杂志

CALLG2008 Protocol is sequential chemotherapy for adult acute lymphoblastic leukemia (ALL) established by Collaborative Group of adults acute lymphoblastic leukemia. It is emphasized that comprehensive treatment of adult ALL according to risk stratification is rather important. This study was purposed to evaluate the therapeutic efficacy of CALLG2008 for adult ALL. The clinical data of adult ALL patients of ≥ 14 years old diagnosed and treated by CALLG2008 Protocol were collected from May 1, 2009 to December 31, 2011 in Fujian Medical University Union Hospital, and the efficacy was analyzed. The results showed that 31 out of 33 cases of ALL achieved CR, the CR rate was up to 93.9%, the PR rate was 3.1%, and the total response rate was 97%. There were no uncontrolled severe toxicities, and no early deaths were observed. The overall survival (OS) at 1 year was only 66.7%,the relapse rate was 43.8% and the 1-year mortality was 33.3 %. This may be related with no-enough compliance, no-enough economical support and short follow-up time of the patients. The risk factor analysis showed that WBC level in newly diagnosed patients may influence the OS and relapse-free survival (RFS) of ALL. It is concluded that CALLG2008 protocol applied to adult ALL has a high remission quality and low mortality rate during the induction. The disease free survival (DFS) needs to be observed longer. It is essential to carry out MRD monitoring to determine the early recurrence and improving the long-term efficacy.

Effect of cucurmosin on chronic myeloid leukemia K562 cell line / 中国实验血液学杂志

This study was aimed to investigate the antitumor effect of pumpkin protein (cucurmosin, CUS) on subcutaneous transplant tumor in chronic myeloid leukemia K562 cell-NOD/SCID mice and leukemia model. The subcutaneous transplant tumor in K562-NOD/SCID mice and leukemia model were established; using two models, the antitumor activity of CUS in mice was evaluated. The results indicated that the inhibitory rate of 0.5 mg/kg and 1 mg/kg CUS on subcutaneous transplant tumor were 53.45% and 59.43% respectively; survival time of mice received 0.25 mg/kg and 0.5 mg/kg CUS was 39.8 ± 5.5 d and 43.4 ± 6.6 d, antitumor rate was 24.9% and 36% respectively. It is concluded that CUS has significant inhibitory effect on mice with CML cell line K562.

Expression and significance of stathmin1 in acute leukemia / 中国实验血液学杂志

This study was aimed to investigate the expression of stathmin1 mRNA and stathmin1 protein in de novo patients with acute leukemia (AL), relapsed patients with AL and complete remission patients with AL, and its clinical significance. The expression of stathmin1 mRNA and stathmin1 protein in peripheral blood samples from 76 cases of AL and 25 healthy persons were examined by fluorescent quantitative PCR (FQ-PCR) and Western blot, respectively. The results showed that the stathmin1 protein expression could not be detected in healthy persons, only the low level of its mRNA could be observed in them. The stathmin1 mRNA expression level in de novo AL patients was higher than that in healthy persons (P < 0.05), the stathmin1 mRNA expression level in relapsed patients with AL was higher than that in de novo patients (P < 0.05), and there was no significant difference of stathmin1 mRNA expression between patients with AML and patients with ALL. The positive rate of stathmin1 protein expression in de novo patients with AL was 89%, while it obviously decreased or did not express in complete remission patients with AL. The stathmin1 protein expression in relapsed patients with AL did not display significant difference as compared with that in de novo patients (P > 0.05). There was no significant difference in stathmin1 protein expression between patients with AML and patients with ALL (P > 0.05). It is concluded that stathmin1 protein and mRNA are overexpressed in de novo patients and relapsed patients, and lowly expressed in complete remission patients. Therefore, the stathmin1 may be a new biological marker for evaluation of minimal residual disease.

Effects of cucurmosin combined with common chemotherapeutics on proliferation and apoptosis of NB4 cells / 中国实验血液学杂志

This study was aimed to investigate the proliferation inhibition and apoptosis induction of cucurmosin (CUS) combined with all trans-retinoic acid (ATRA) or arsenic trioxide (ATO) on human acute promyelocytic leukemia cell line NB4. MTT method was used to determine the proliferative inhibition of CUS combined with ATRA or ATO on NB4 cells, and flow cytometry was used to determine the apoptosis induction effect of CUS combined with ATRA or ATO on NB4 cells. Jin's formula was used to assess the synergistic effect of this combinations. The results showed that, compared with single drug, the proliferation inhibitory ratio and apoptotic ratio of CUS combined with ATRA or ATO on NB4 cells was higher than CUS, ATRA and ATO alone. The synergistic index (q) were all larger than 0.85, and the combined effects were significant at low concentrations. It is concluded that the CUS combined with ATRA or ATO synergistically increases the effects of proliferative inhibition and apoptosis induction on NB4 cells.

Efficacy of remission induction chemotherapy and prognostic analysis in elderly patients with acute myeloid leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the outcome of remission induction chemotherapy (IC) and prognostic in elderly patients with acute myeloid leukemia (AML).</p><p><b>METHODS</b>The clinical data of 156 AML patients older than 60 years in the Institute of Hematology, Union Hospital of Fujian Medical University from January 2003 to July 2010 were analyzed retrospectively. 104 patients received cytarabine-based regimens, including protocol DA,IA or CAG,while 52 patients received palliative treatment. The median survival time was compared between patients with and without IC. The prognostic factors were evaluated by using univariate and multivariate analyses.</p><p><b>RESULTS</b>145 (93%) cases were followed-up. The median survival time was 316 days in 96 IC patients, compared with 37 days in 49 PT patients (P < 0.01). Not receiving induction chemotherapy,high-risk karyotype,hyperleukocytosis (> or = 100 x 10(9)/L), Charlson Comorbidity Index (CCI) > or = 2 were adverse prognostic factors of the survival time with univariate analysis, and all were independent poor factors affecting the survival time with multivariate analysis.</p><p><b>CONCLUSIONS</b>IC can improve outcomes in elderly AML patients. The patients with hyperleukocytosis (> or = 100 x 10(9)/L) , high-risk karyotype, CCI > or = 2 and without receiving IC have poorer prognosis.</p>

Apoptosis-inducing effect of C-MYC siRNA on acute lymphoblastic leukemia Jurkat cell line / 中国实验血液学杂志

The study was purposed to investigate the effects of C-MYC siRNA on the proliferation and apoptosis of acute lymphoblastic Jurkat cell line. siRNA targeting the site 1545-1565 of C-MYC mRNA was designed and chemically synthesized, then C-MYC siRNA was transfected into Jurkat cells by the transfer agent (HiPerFect Transfection Reagent), the morphological changes were observed under inverted microscope; the tetrazole compound (MTS) was applied to draw the cell growth curve; the cell colony test was used to detect the effect of C-MYC siRNA on the proliferation of Jurkat cells; the flow cytometry and TUNEL method were used to analyze the apoptosis of Jurkat cells. The results showed that after Jurkat cells were treated with different concentrations of C-MYC siRNA, the growth of Jurkat cells was inhibited to various degrees, inhibitory rate was enhanced as C-MYC siRNA concentration increased. C-MYC siRNA also could obviously inhibit the cell clony formation. The apoptosis of cells could be detected by flow cytometry and TUNEL method, the apoptosis rate of cells increased along with prolonging of treatment with C-MYC siRNA. It is concluded that the chemically synthesized C-MYC siRNA can inhibit significantly the proliferation and induce the apoptosis of Jurkat cells.

Up-regulation of Stathmin and CrkL protein expressions in adriamycin-resistant leukemia cell line K562/A02 / 中国实验血液学杂志

The purpose of this study was to compare the differences of the protein expression profiles between human myeloid leukemia K562 cells and adriamycin-resistant K562/A02 cells, as well as to select novel resistance-related proteins in myeloid leukemia by means of proteomics. The total cellular proteins were separated from K562 and adriamycin-resistant K562/A02 cells by using technique of two dimensional difference in gel electrophoresis (2D-DIGE). Differentially expressed proteins were analyzed by matrix-assisted laser desorption ionization/time of flight-mass spectrometry (MALDI-TOF/MS), and by protein database searching. Moreover, the differentially expressed proteins were verified at protein and mRNA levels by Western blot assay and quantitative real time PCR. The results showed that 8 proteins differentially expressed in adriamycin-resistant K562/A02 cells, among them 2 proteins were identified to be down-regulated and 6 to be up-regulated. These identified proteins involved in the cell energy metabolism, cell proliferation, cell apoptosis, signal transduction, gene transcription and translation respectively. The results assayed by Western blot were similar to those detected by 2D-PAGE. Two up-regulated proteins Stathmin and CrkL were selected for verification in K562 and K562/A02 cells. As a result, the results detected by Western blot were identical with results from 2D-DIGE; real time quantitative PCR assay showed that the changes of CrkL at mRNA level were identical with changes at protein level, but no complete identity of Stathmin changes at mRNA level and protein level was observed. It is concluded that the difference of protein expression profile exists in K562 and K562/A02 cells. Stathmin and CrkL proteins may be involved in the drug resistance and suggest a novel clue for the resistant mechanisms in myeloid leukemia, which is worth further to explore.

Effects of small interference RNA against c-myc on expression of c-myc and h-tert genes in acute lymphoblastic leukemia cell line Jurkat / 中国实验血液学杂志

This study was purposed to investigate the effect of small interfering RNA against c-myc on c-myc, h-tert gene and protein expressions in acute lymphoblastic leukemia cell line (Jurkat cells), so as to provide new methods and targets for gene therapy of leukemia. The siRNA against target sites 1545-1565 of c-myc mRNA was chemically synthesized and was transfected into Jurkat cells by transfectant. The c-myc, h-tert mRNA and protein expression levels before and after treatment with c-myc siRNA were detected by RT-PCR and Western blot, respectively. The results showed that c-myc siRNA obviously inhibited the proliferation of Jurkat cells, the half inhibitory concentration (IC50) for 48 hours was about 75 nmol/L. c-myc siRNA could decrease c-myc, h-tert mRNA and C-MYC, h-TERT protein expression levels of Jurkat cells. It is concluded that c-myc siRNA significantly reduce c-myc, h-tert mRNA and protein expression levels through inhibiting c-myc mRNA expression and decreasing intracellular level of C-MYC protein.

Effect of emodin on induction of apoptosis in jurkat cells and its possible mechanisms / 中国实验血液学杂志

The aim of study was to investigate the effect of a traditional Chinese medicine, emodin, on proliferation and apoptosis in T lymphocytic leukemic cell line Jurkat and its mechanisms. Cell proliferation inhibition was detected by MTT assay. Cell apoptosis was measured by DNA ladder and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. The expressions of related proteins and caspase family members were determined by Western blot. The results showed that emodin inhibited proliferation in Jurkat cells, with an IC50 about 20 micromol/L and induced cell apoptosis in both time-and dose-dependent manners. The expressions of proliferation-related protein C-MYC, hTERT and apoptosis-related protein BCL-2 were down-regulated in a time dependent manner after the treatment with emodin. The expressions of procaspase-3, -8 and -9 all decreased while activated caspase-3 and PARP expressions were up-regulated. It is concluded that emodin can remarkably inhibit cell proliferation and induce apoptosis in Jurkat cells. The down-regulation of proliferation-related proteins C-MYC, hTERT and apoptosis-related protein BCL-2 expressions and activation of caspase cascade may be involved in the process of apoptosis.

Apoptosis of human myeloid leukemia cell line HL-60 cells induced by siRNA targeting gene c-myc / 中国实验血液学杂志

This study was purposed to investigate the effects of siRNA targeting c-myc on apoptosis induction, proliferation in inhibition as well as c-myc protein and mRNA expression in human myeloid leukemia cell line HL-60 cells. C-myc siRNA synthesized in vitro was transfected into HL-60 cells by liposome. Changes of cell morphology were observed. Growth inhibition was detected by MTT assay and colony formation assay, and cell apoptosis was determined by DNA ladder. The expressions of c-myc mRNA and protein were detected by RT-PCR and Western-blot respectively. The results indicated that c-myc siRNA remarkably inhibited the cell proliferation, with an IC(50) value of 150 nmol/L. Data of DNA ladder showed that HL-60 cells apoptosis could be efficiently induced by c-myc siRNA, the apoptosis rate positively correlated with the time duration of treatment with drugs. The c-myc mRNA and protein expressions on HL-60 cells decreased after treatment with c-myc siRNA, which negatively correlated with time duration of treatment. It is concluded that c-myc siRNA can efficiently induce growth inhibition, decrease the expressions of c-myc mRNA and protein, and induce apoptosis in HL-60 cells.

Overview of chronic myelogenous leukemia and its current diagnosis and treatment patterns in 15 hospitals in China / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore demographic characteristics, current diagnosis and treatment patterns of chronic myelogenous leukemia (CML) patients in China.</p><p><b>METHODS</b>Data of hospitalized CML patients in 2005 whole year and outpatient information (July 1 through September 30, 2006) from 15 hospitals throughout China were analyzed.</p><p><b>RESULTS</b>A total of 1824 CML cases were analyzed, including 722 inpatients and 1102 outpatients. The male/female ratio was 1.78:1. The median age at diagnosis was 40.02 (2.45 - 83.29) years old, 90.41% of the patients were diagnosed at chronic phase. Proportion of accelerated phase or blast crisis patients increased to 21.66% during study period. 93.20% of the patients received blood routine and bone marrow morphologic examination at diagnosis and in monitoring; 70.29% were performed cytogenetic analysis and 51.54% performed molecular measurement in addition. The most common therapy for CML treatment was hydroxycarbamide. The proportion of patients treated with imatinib and interferon was 37.45% and 25.55%, respectively. Of 722 inpatients, 164 (22.72%) received hemotopoietic stem cell transplantation (HSCT). The proportions of accelerated phase and blast crisis patients treated with imatinib were 48.28% and 48.42%, respectively, being significantly higher than that of chronic phase patients (35.9%) (P < 0.05). The mean imatinib dosage administered in the three phases patients did not differ significantly. Imatinib resistance rates were 6.87% and 16.28% for outpatient and inpatient, respectively. In the outpatient group, the primary resistance to imatinib occurred comparably to the secondary resistance (68.75%), while primary resistance was predominant in inpatient group (65.71%). The intolerance rates of imatinib for outpatient and inpatient were 3.21%, 11.63%, respectively. The majority of patients treated with imatinb were not monitored in time: 63.38% patients evaluated hematologic response after 3 months of treatment, proportions of patients received cytogenetic examination after 6 months and 12 months of treatment were 41.41% and 27.35%, respectively. Mean cost for HSCT was 213 092 +/- 125 890 RMB.</p><p><b>CONCLUSIONS</b>CML in China tends to afflict younger population than in Western countries. Most patients were diagnosed in the chronic phase. Due to restriction of financial support, only one third of CML patients were treated with imatinib, and the majority of the treated were not monitored in time. Clinicians should pay attention to resistance and intolerance to imatinib treatment in accelerated phase or blast crisis patients.</p>

Effects of zhiling capsule on the proliferation inhibition and apoptosis induction in human small cell lung cancer cell line NCI-H446 / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effects of Zhiling Capsule (ZLC) on the proliferation inhibition and apoptosis induction in human small cell lung cancer cell line NCI-H446.</p><p><b>METHODS</b>According to the different components of ZLC, NCI-H446 cells were treated with traditional Chinese medicine, Western medicine and ZLC compound groups. The rates of cell viability and colony formation were observed by MTT assay and colony formation assay respectively. Cell cycle assay, Bcl-2 protein expression, chondrial transmembrane potential and Caspase-3 activity were detected by flow cytometer. Apoptotic cells were detected by DNA fragmentation assay, Annexin-V FITC staining and TUNEL labeling methods.</p><p><b>RESULTS</b>After NCI-H446 cells were treated with various concentrations of drug groups, cell growth was significantly inhibited in a dose dependent manner. Cell colony formation was obviously lowered in the same way. The levels of chondrial transmembrane potential and Bcl-2 protein expression were decreased, while the levels of Caspase-3 activity were increased after the treatment. Typical DNA ladder were seen from gel electrophoresis, and apparent apoptotic peaks were observed by flow cytometer. Apoptosis occured in the early and late stage was identified by Annexin-V FITC staining and TUNEL labeling methods respectively. The ZLC compound group has stronger apoptosis induction than the other groups.</p><p><b>CONCLUSION</b>ZLC could efficiently inhibit growth and induce apoptosis in NCI-H446 cells, which may be related with the down-regulation of chondrial transmembrane potential and Bcl-2 protein expression and the up-regulation of Caspase-3 activity.</p>

c-myc Antisense Oligodeoxynucleotides Induce Apoptosis of Human Myeloid Leukemia Cell Line HL-60 / 中国实验血液学杂志

In order to study the effects of c-myc antisense phosphorothioate oligodeoxynucleotide in inducing apoptosis of HL-60 cells, the expression of c-myc mRNA was determined by RT-PCR, the morphologic signs of apoptotic cells were observed by transmission electron microscopy, the proportion of apoptotic cells was detected by flow cytometry, and the DNA fragments were analysed by agarose gel electrophoresis. Results of RT-PCR showed marked decrease of c-myc mRNA expression in AspoI and II treated cells. The level of c-Myc protein was decreased 23.8% and 45.4%, respectively, in cells treated with 5 and 10 micro mol/L AspoI, and 38.4% after treatment of 10 micro mol/L AspoII. The apoptotic rates were 23.97% and 52.6% after 10 micro mol/L AspoI treatment and 28.8% and 45.19% after treatment with AspoII 10 micro mol/L for 48 and 72 hours, respectively. while apoptotic cells did not apear in control and sense oligodeoxynucleotide groups. Electron microscopy observation showed the characteritics of apoptosis. A ladder like pattern of DNA fragments was demonstrated on electrophoretogram. The results suggst that c-myc antisense oligodeoxynucleotides could be highly specific gene agents that can suppress the level of c-myc mRNA, decrease the c-Myc proteins and induce apoptosis of HL-60 cells.