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Objective@#To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison.@*Methods@#Peking University People’s Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients. Totally 23 sets with 14 different samples per set were prepared. TRIzol reagent was added in each tube and thoroughly mixed with cells for homogenization. Each laboratory simultaneously tested RUNX1-RUNX1T1 and WT1 transcript levels of one set of samples by real-time quantitative PCR method. All transcript levels were reported as the percentage of RUNX1-RUNX1T1 or WT1 transcript copies/ABL copies. Spearman correlation coefficient between the reported transcript levels of each participated laboratory and those of PKUPH was calculated.@*Results@#①RUNX1-RUNX1T1 comparison: 9 samples were (+) and 5 were (-) , the false negative and positive rates of the 20 participated laboratories were 0 (0/180) and 5% (5/100) , respectively. The reported transcript levels of all 9 positive samples were different among laboratories. The median reported transcript levels of 9 positive samples were from 0.060% to 176.7%, which covered 3.5-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.5 to 12.3 (one result which obviously deviated from other laboratories’ results was not included) , 85% (17/20) of the laboratories had correlation coefficient ≥0.98. ②WT1 comparison: The median reported transcript levels of all 14 samples were from 0.17% to 67.6%, which covered 2.6-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.3-13.7, 62% (13/21) of the laboratories had correlation coefficient ≥0.98. ③ The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of WT1 transcript levels. Both RUNX1-RUNX1T1 and WT1 transcript levels from 2 and 7 laboratories were individually lower than and higher than those of PKUPH, whereas for the rest 11 laboratories, one transcript level was higher than and the other was lower than that of PKUPH.@*Conclusion@#The reported RUNX1-RUNX1T1 and WT1 transcript levels were different among laboratories for the same sample. Most of the participated laboratories reported highly consistent result with that of PKUPH. The relationship between laboratories of the different transcript levels may not be the same.
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Objective To investigate the role of Th17 cells in chronic myeloid leukemia (CML) pathogenesis. Methods 33 CML patients [15 newly diagnosed (ND)- and 18 chronic-phase (CP)- CML patients] and 15 healthy controls were enrolled. The percentage of Th17 cells in the peripheral blood (PB) of CML and controls were evaluated by flow cytometry. RORC mRNA expressions were examined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). The levels of IL-17 in PB of CML and controls were measured by enzyme-linked immunoadsorbent assay (ELISA). The relationships between Th17 cells and clinical characteristics in CML were analyzed. Results The percentage of Th17 cells in PB of ND-CML patients [(0.71±0.41) %] was significantly lower than that of CP-CML patients [(4.08±0.74) %, P<0.05] and healthy controls [(3.18±1.32) %, P<0.05]. The mRNA level of RORC in PB of ND-CML patients (0.043 3±0.040 5) was significantly decreased compared with that in CP-CML patients (0.086 1±0.052 3, P<0.05) and healthy controls (0.091 0±0.058 4, P<0.05). The levels of IL-17 in PB of ND-CML patients [(1.43±0.22) pg/ml] and CP-CML patients [(1.36±0.19) pg/ml] were slightly higher than that of healthy controls [(1.23±0.14) pg/ml, P< 0.05]. The percentage of Th17 cells had significantly negative correlations with white blood cell counts in PB or bcr-abl (IS). Conclusion Th17 cells may play an important role in CML pathogenesis, which has potential implication for immunotherapy of this malignancy.
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<p><b>OBJECTIVE</b>To detect JAK2 V617F mutation burden and its clinical implications in patients with myeloproliferative neoplasm (MPN).</p><p><b>METHODS</b>JAK2 V617F mutation burden were detected by using MGB Taqman probes and its clinical significance were retrospectively studied in 415 MPN patients.</p><p><b>RESULTS</b>JAK2 V617F was found in 56.9% of all patients [83.5% in polycythemia vera (PV), 55.9% in essential thrombocythemia (ET), 41.9% in primary myelofibrosis (PMF) and 64.7% in MPN-unclassifiable)]. The majority of patients carried heterozygous JAK2 V617F mutation and homozygote was found only in 12 cases (4 in PV, 4 in MPN-U, 2 in PMF, 1 in ET, and 1 in chronic neutrophilic leukemia). Most patients (68.8%) were lower mutation burden (mutation burden<50%), but PV had the highest burden, the moderate burden in PMF and the least in ET. The patient's age and WBC count were significantly correlated with higher mutation burden in PV. WBC count was significantly related to higher mutation burden in ET. WBC count, Hb level and the platelet count were significantly related to higher mutation burden in PMF.</p><p><b>CONCLUSION</b>The mutation burden of JAK2 V617F from high to low was PV, ET and PMF. The majority of JAK2 V617F mutation was heterozygous. JAK2 V617F mutation burden was positively correlated with age, WBC, Hb and platelet counts.</p>
Subject(s)
Humans , Homozygote , Janus Kinase 2 , Leukocyte Count , Mutation , Myeloproliferative Disorders , Platelet Count , Polycythemia Vera , Retrospective Studies , Thrombocythemia, EssentialABSTRACT
<p><b>OBJECTIVE</b>To revalidate the conversion factor（CF）for the conversion of BCR-ABL （P210）transcript levels to the international scale（BCR- ABLIS）in chronic myeloid leukemia（CML） which validated before.</p><p><b>METHODS</b>Peking University People's Hospital（PKUPH）prepared the exchange samples for revalidation of CFs of 15 laboratories which validated nine or eighteen months ago. The fresh BCR-ABL（P210）（+）bone morrow or peripheral blood nucleated cells were diluted with BCR-ABL （P210）（-）cells to achieve different BCR- ABL levels, totally 16 sets and 24 samples per set were prepared. TRIzol reagent was added in each tube. Each laboratory tested BCR-ABL transcript levels of one set of samples. Agreement between BCR-ABLIS of each laboratory and PKUPH was assessed by the Bland- Altman method. For laboratories which did not meet the criteria of revalidation, linear regression equation was derived after the samples with maximum BCR-ABL deviation were removed until R²>0.98, then new CF was calculated.</p><p><b>RESULTS</b>10 laboratories met the revalidation criteria with both bias within ±1.4 fold and 95% limits of agreement within ±6 folds, and their CFs still could be used for accurately conversion of BCR-ABLIS. New CFs were recalculated as of 1.8-6.3 folds of their previous CFs in 5 laboratories not met the criteria.</p><p><b>CONCLUSION</b>Revalidation of CF by sample exchange among laboratories was necessary for accurate and continuous application of BCR-ABLIS, which not only tested the validity of CF acquired before but also calculated new available CFs for those with invalid CFs.</p>
Subject(s)
Humans , Bone Marrow Cells , Fusion Proteins, bcr-abl , Genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Diagnosis , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To validate the conversion factor (CF) for the conversion of BCR-ABL (P210) transcript levels to the international scale in chronic myeloid leukemia (CML).</p><p><b>METHODS</b>In 2012, the international reference laboratory in Adelaide, Australia (IMVS) sent two batches of RNA samples, 30 samples per batch, to Peking University People's Hospital (PKUPH). By comparing BCRABL (P210) transcript levels reported by the two laboratories, CF of PKUPH was calculated and validated by IMVS. In 2013, PKUPH prepared the exchange samples for validation of CF of 9 hospitals who have calculated CFs before. The fresh BCR-ABL (P210) (+) cells were serially diluted by BCR-ABL (P210) (-) cells to prepare 22 kinds of samples with different BCR-ABL transcript levels, each kind had 10 parallel samples. Trizol reagent was added in each tube. Ten hospitals tested BCR-ABL transcript levels of one set of 22 samples. Agreement between BCR-ABL transcript levels of each laboratory and PKUPH was assessed by the Bland-Altman method.</p><p><b>RESULTS</b>PKUPH successfully validated its CF with bias 1.1 fold and 95% limits of agreement between -4.7 and 4.9 fold. Of 9 hospitals whose validation performed by sample exchanges with PKUPH, 6 hospitals successfully validated their CF with bias ≤±1.4 fold and 95% limits of agreement within ±6 fold.</p><p><b>CONCLUSION</b>Validation of CF examined the stability of the detection of BCR-ABL (P210) transcript levels, which was necessary for the valid conversion of BCR-ABL (P210) transcript levels to the international scale in CML.</p>
Subject(s)
Humans , Fusion Proteins, bcr-abl , Genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Reference Standards , Transcription, GeneticABSTRACT
Objective To address the standard first-line management under the new diagnostic criteria in adult immune thrombocytopenia (ITP). Methods A retrospective analysis was conducted involving 178 adult ITP patients treated with high-dose dexamethasone or prednisone in Qilu Hospital from March 2004 to November 2009 using new diagnostic criteria. Results The median age was 41 years with a male/female ratio of 0. 73: 1. Among the 178 ITP patients, 87 were newly diagnosed, 30 persistent ITP, 58 chronic ITP, and 3 unable to follow up. The efficacy rates among 167 patients able to assess in the three groups were 77.4% ( 65/84 ), 64. 0% ( 16/25 ) and 62. 1% ( 36/58 ) respectively, and their complete remission (CR) rates were 57. 1% (48/84), 36. 0% (9/25) and 32. 8% (19/58). The efficacy rate and CR rate of the newly diagnosed ITP category were significantly higher than those of the chronic ITP category (x2 = 3. 917, P ＜ 0. 05 ;x2 = 8. 186, P ＜ 0. 01 ). The patients treated with high-dose dexamethasone or prednisone therapy had no significant differences in sex, age or blood platelet count before treatment.Moreover, the short or long term response rates and the CR rates between the two therapies had no statistically significant differences while the former had a shorter onset time ( F = 10. 34, P ＜ 0. 01 ).Conclusions The study sets up a basis for the application of the recommended new definition and outcome criteria for adult ITP. Dexamethasone therapy is favored as first-line therapy.
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Objective To construct an eukaryotic expression vector of retinoblastoma gene (pIRES-Rb94), and investigate the radiosensitising effect of Rb94 on lung adenocarcinoma cell line A549 and the mechanism. Methods Recombinant expression plasmid pIRES-Rb94 was constructed and then transfected into A549 cells using lipofectamine 2000. Steadily transfected cells were obtained using G418 se-lecting system. Cell counting method was used to produce the growth curve and the population doubling time was then calculated. The radiosensitivity of A549 cells was assessed by clonogenic assay. The expression of bTERT and Bcl-2 mRNA was detected by real-time RT-PCR. Cell cycle distribution was measured by flow cytometry. Results Steadily transfected cell line pIRES-Rb94-A549 was aquired. Compared with A549 cells, the population doubling time of pIRES-Rb94-A549 cells was increased from 31.5 h to 39.5 h (t=5.15, P<0.01). The expression of hTERT and Bcl-2 mRNA was both down-regulated (0.02:1.00, t= 18.99,P<0.01,0.01:1.00,t=13.73,P<0.01). The number of cells was increased in G2/M phases (35.91%:4.53%, t =36.78,P=0.00), whereas decreased in G0/G1 and S phases (47.02%:74.07%, t =11.71,P=0.00;17.07%:22.32%, t =2.30,P<0.05). The sensitizing enhancement ratio was 1.30. Conclusions Rb94 has marked radiosensitizing effect on A549 cells by G2/M phase blockage and down-regulation of hTERT and Bcl-2 mRNA expression. Rb94 may also inhibit the ability of cell proliferation by regulating cell cycle distribution.
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<p><b>OBJECTIVE</b>To investigate the percentage of splenic CD(5)(+) B lymphocytes in chronic idiopathic thrombocytopenic purpura (IT) and the impact of splenic CD(5)(+) and CD(5)(-) B lymphocytes on the production of platelet glycoprotein (GP)-specific autoantibodies.</p><p><b>METHODS</b>Splenic CD(5)(+) B lymphocytes were identified by two-color flow cytometric analysis in eight patients. Four of the eight patients displayed plasma autoantibodies against both GPIIb/IIIa and GPIb/IX, and their splenic B lymphocytes were separated by Ficoll-Hypaque density gradient and sheep erythrocyte, and further purified by magnetic activate cell separation (MACS). Purified CD(5)(+) and CD(5)(-) B lymphocytes were cultured separately with or without staphylococcus aureus cowan I (SAC). GP specific autoantibodies in culture supernatants were measured by modified monoclonal antibody immobilization of platelet antigen assay (MAIPA).</p><p><b>RESULTS</b>The percentage of splenic CD(5)(+) B lymphocytes in ITP patients was slightly higher than that in control with no statistical significance. MACS purified splenic CD(5)(+) and CD(5)(-) B lymphocytes from three out of four ITP patients produced high levels of anti-GPIIb/IIIa and anti-GPIb/IX antibodies. Culture supernatants of CD(5)(+) B lymphocytes from the other patient showed positive reaction only in GPIb/IX MAIPA. Culture supernatant of CD(5)(-)B lymphocytes from the same patient were double positive in both GPIIb/IIIa and GPIb/IX MAIPA.</p><p><b>CONCLUSIONS</b>Both splenic CD(5)(+) and CD(5)(-) B lymphocytes produce platelet GP-specific autoantibodies in chronic ITP with similar antibody spectrum and titer, and may all play a role in the autoimmune pathogenesis of ITP.</p>
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Adolescent , Adult , Female , Humans , Male , Middle Aged , Autoantibodies , B-Lymphocytes , Allergy and Immunology , CD5 Antigens , Chronic Disease , Platelet Glycoprotein GPIIb-IIIa Complex , Allergy and Immunology , Platelet Glycoprotein GPIb-IX Complex , Allergy and Immunology , Purpura, Thrombocytopenic, Idiopathic , Allergy and Immunology , Spleen , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To explore the mechanism of Notch signaling transduction system and its effects on hematopoietic system.</p><p><b>METHODS</b>Notch ligands transfected CHO cells were added into Notch1 and Notch2 transfected CHO cells, which were transiently transfected with reporter gene TP1. PGL-100 was used as substrate to test the interaction between Notch and Notch ligands. CHO, Jagged2-CHO and Delta 4-CHO cells were seeded in the petri dish containing G-CSF, and then Notch 1-32D cells were added in it to observe the differentiation of Notch1-32D cell after incubation and staining.</p><p><b>RESULTS</b>All of the five Notch ligands binding to Notch1 could induce TP1 activity, it increased significantly the Jagged2-CHO, Delta 4-CHO1-4 and Delta 4-CHO1-5 cells. For Notch2, the TP1 activity induced by the five ligands in these cells was much higher than that of CHO. At the presence of G-CSF, Notch1-32D could differentiate to mature granulocyte. Jagged2 could inhibit G-CSF induced Notch1-32D cell differentiation, but Delta 4 could not.</p><p><b>CONCLUSION</b>Jagged2 and Delta 4 are the ligands of Notch1. Jagged2 can inhibit G-CSF induced Notch1-32D cell differentiation, but Delta 4 can not.</p>
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Animals , Cricetinae , CHO Cells , Calcium-Binding Proteins , Genetics , Metabolism , Physiology , Cell Differentiation , Physiology , Cricetulus , Intercellular Signaling Peptides and Proteins , Genetics , Metabolism , Physiology , Membrane Proteins , Genetics , Metabolism , Physiology , Receptor, Notch1 , Genetics , Metabolism , Physiology , Receptor, Notch2 , Genetics , Metabolism , Physiology , Receptors, Notch , Genetics , Metabolism , Physiology , Serrate-Jagged Proteins , Signal Transduction , Physiology , TransfectionABSTRACT
Objective:To assess the potential of mouse immune-costimulating signal B7-2 in inducing immune effect.Methods:(1)The specific mB7-2cDNA fragment from LPS-stimulated mouse splenocytes was obtained by reverse transcription and polymerase chain reaction.The obtained fragment was inserted to pLXSN plasmid.Then the plasmid pLXSN-mB7-2 was packaged into PA317 cells;(2)The EL4/mB7-2 was obtained by infecting the EL4 by the concentrated virus particles produced by PA317/mB7-2;(3)In vitro,the secretion of IL-2 of EL4/mB7-2 stimulated-lymphocytes was detected by using mixed lymphocytes culture.Results:pLXSN-mB7-2 and transgenetic EL4/mB7-2 cells were obained successfully.IL-2 production in 24h in the supernatant stimulated by EL4/mB7-2 was much higher than wild EL4 cells.Conclusion:EL4/mB7-2 can activated T cell to produce IL-2.This research lay foundation for the research of the function of immune-costimulating signal in the tumor immunity and treatment,the mechanism of autoimmune disease and organ transplantation.
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Objective To investigate cell-killing and bystander effects on gallbladder cancer (GBC-SD) cells transferred with double suicide genes (CD and HSV-tk). Methods CD and HSV-tk genes were transfected into PA317 cells using lipofectamine. GBC-SD cells were infected with viral supernatant. GBC-SD/CD+tk cells were treated with 5-FC and/or GCV. GBC-SD/CD+tk and GBC-SD were inoculated subcutaneously into the nude mice respectively and treated with GCV and 5-FC for 21 days. Results The killing effect of 5-FC and GCV in combination on GBC-SD/CD+tk cells is more effective than using 5-FC or GCV alone. The local bystander effect is significant while the distant bystander effect was not obvious. Conclusion The cell-killing efficacy of chemotherapy on GBC-SD transfected with double suicide gene was significantly increased, and the bystander effect was obvious.
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Objective To investigate the repressing effects of cytosine deaminase(CD) and herpes simplex(virus) thymidine kinase(HSV-tk) double suicide genes coexpression system on human cholangiocarcinoma QBC939 cells and QBC939 cells transplantated tumor in nude mice.Methods CD and HSV-tk double suicide genes were transfered into QBC939 cells using liposomes.After G418 selection,the positive clones of QBC939/CD+tk cells were picked up and cultured.The expression of CD and HSV-tk genes was(confirmed) by RT-PCR.In vitro,the QBC939/CD+tk cells were treated with 5-Fc and/or GCV,and the cytoxicity efficacy was evaluated by microculture tetrajolium test(MTT) method.The QBC939/CD+tk cells were inoculated subcutaneously into nude mice,and when the tumors were palpable,5-Fc and GCV were(injected) intraperitoneally,and the volumes of transplantated tumors were measured before and after medication.Results Double suicide genes were stably expressed in QBC939/CD+tk cells.The repressing capability of combination of 5-Fc and GCV on QBC939/CD+tk cells was more effective than that of using either 5-Fc or GCV alone.The increase of cell-repressing was assaciated with increase the concentration of the prodrug.The repressing effect of combination of the 2 prodrugs on early period of transplantation tumor was obvious,even complete abolition of tumor was noted,and moreover a marked local bystander effect was observed.(Conclusions) In vitro and in vivo,the cell-repressing efficacy of double suicide gene system on cholongiocarcinoma cells and the tramsplanted tumor of nude mice was significant,and the bystander effect was obvious.
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Purpose: To investigate the effect of HSV-TK/GCV system on human prostate cancer. Methods: MTTassay, flow cytometry( FCM), optical and electron microscopy were used to determine the sensitivity of GCV on prostate cancer cell( PC-3m) after being tranfected with HSV-TK gene. Results: It was found that GCV had significant cytotoxic activity on HSV-TK gene-transduced prostate cancer cells, but little effect on the non-transduced cells. Conclusions: The experiment indicates that HSV-TK/GCV system has significant antitumor activities on the TK gene-transduced prostate cancer cells in vitro.
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Objective:To investigate the different killing effects on human hilar cholangiocarinoma cells FRH with cytosine deaminase(CD) and herpes simplex virus thymidine kinase(HSV tk)double suicide genes coexpression compared with single gene mediated by retrovirus.To find a more efficient and low toxicity suicide gene therapy for hilar cholangiocarinoma.Methods:CD and HSV tk double suicide genes were transfected into PA317 cells using lipofectamine.The positive clones were picked out and cultured after G418 selected.The viral supernatant was collected.The FRH cells were infected with the virus containing the double suicide genes.After G418 selection,RT PCR was resorted to demonstrated the successful transcription of CD and HSV tk genes.The FRH/CD+tk and FRH cells in culture were respectively treated with 5 Fc and /or GCV.The cytoxicity efficacy was evaluated by microculture tetrajolium test (MTT) method.Results:The virus containing double suicide gene was produced in PA317 cells.Double suicide genes were stably expressed in RFH cells after being infected with the virus.The killing effect of combination 5 Fc with GCV on FRH/CD+tk cells is more effective than that of using 5 Fc or GCV alone.Conclusion:The CD+TK/5 Fc+GCV co expression system is more effective for killing effects on FRH cells than that of CD/5 Fc or tk/GCV system alone.
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Objective To study the in vitro effects of sonodynamic management on human ovarian cancer cell line HRA. Methods Hematoporphyrin was selected as a sonosensitizer and ultrasound with certain intensity was used to activate hematoporphyrin.Then MTT assay and clony-forming efficiency assay were applied to determine the growth inhibitory effects of ovarian cancer cells. Electron microscope was applied to detect the morphological changes of the cells. Cell apoptosis was analyzed using flow cytometry. Results Hematoporphyrin had no significant inhibitory effects on the cells. However, when hematoporphyrin was used before ultrasound,it could enhance the killing effects of ultrasound(P
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Objective: To investigate effect of exogenous wild-type p53 gene on the cell growth and tumorigenicity of human gallbladder cancer cell lines. Methods: After identification of the genetic status of p53 gene of GBC-SD cell lines with the immunocytochemistry staining and the direct sequencing technique of PCR products, eukaryotic expressing plasmid pCMV-p53 was introduced by lipofectamine-mediated into GBC-SD cell lines. Growing transfected cells were selected by G418. The presence and expression of exogenous p53 gene was detected by PCR, RT-PCR and Western blot. The cellular proliferating ability was assessed using the cell growth curve and cloning assay. The xenograft in nude mice was performed to examine the effect of tumorigenicity. Results: P53 protein overexpression was showed in GBC-SD cell lines. A transversion of TAC→AAC at codon 126 of exon 5 was confirmed. PCR, RT-PCR and Western blot showed exogenous p53 gene had successfully transfected into GBC-SD cells and obtained high expression. The growth and proliferation of the cells were greatly decreased, and the tumorigenicity was significantly inhibited after transfection wtp53. Conclusion: The expression of exogenous wild-type p53 gene could effectively inhibit the growth of gallbladder cancer GBC-SD cells in vitro and in vivo.
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Objective: To explore the enhanced cell killing effect of HSV tk using VP22 intercellular traffciking. Methods: The chimeric genes were constructed by fusing a marker gene for the green fluorescent protein (GFP) or a prodrug enzyme gene for the Herpes simplex virus thymidine kinase (HSV tk) with that of VP22. After being sequenced, the fusion genes were transferred into 293T or COS7 cells. The transfection efficiency and intercellular trafficking were certified using Western blot and immunofluorescence.The cell proliferation was detected through MTT method in the different concentration of GCV and under indicated between transfected cells and untransfected cells. The supernatant of transfected cells was used to culture the untransfected cells to test whether the bystander effect could transferred by media. Results: The gene insertion was proved correct using PCR and DNA sequencing. When the fusion genes were transferred into 293T or COS7 cells at transfection efficiency of 25%～30%, fusion proteins were expressed and efficient intercellular trafficking was demonstrated.The VP22 HSV tk, as a prodrug enzyme fused with VP22, showed an amplified cell killing effect in the presence of GCV as low as 0.1 ?g/ml. Further quantification of the bystander effect showed that cell killing increased with higher proportion of VP22 HSV tk expressing cells. The bystander effect could not be transferred through media. Conclusion: These results clearly indicate that VP22 enhanced intercellular trafficking promotes tumor cell killing effect of HSV tk/GCV.
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Objective To explore the effect of sodium fluoride(NaF) on the tumor markers of human embryonic bronchial epithelium (HEBE) cells.Methods HEBE cells were collected from the human abortive fetues.The cells were exposed to NaF of several concentrations for 36 h.After rinsed,the cells were incubated for 36 h again.The NaF toxicity to HEBE cells was detected using MTT method.The supernatant was collected and the lung tumor markers were detected using ELISA method.Results NaF was toxic to the HEBE cells,and the toxicity was increased with the NaF doses.There were few survival HEBE cells at 6 mmol/L NaF.The tumor markers in both of the control and experiment groups were very low,and no significant difference had been seen between them.Conclusion NaF may damage HEBE cells,but may not influence the tumor markers of HEBE cells.