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Volume 16,Issue 5,2009 Table of Contents
2009, 16(5):427-430.
DOI: 10.3872/j.issn.1007-385X.2009.5.001
Abstract:
New tumor antigens are continuously being discovered due to the improvement of immunologic techniques. Whether dendritic cells induce immune activation or immune inhibition after they capture tumor antigens depends on the danger signals (GMCSF, MCP1, and HSP) or the inhibitory signals (TGFβ, IDO, and iNOS) released by tumor cells. Under the regulation of danger signals, dendritic cells activate Th1 immune response and eliminate tumors; under the regulation of inhibitory signals, they activate Th2 immune response and can not effectively eliminate tumors. Progress in tumor immunotherapy mainly is manifested by antibodybased therapy, T cellbased therapy, and tumor vaccinebased therapy. To day at least 7 antibodies have been confirmed effective when combined with chemotherapeutic agents in treatment of tumors. Despite of the progress made in antibody therapy, discovery of new targets, development of new antibodies, and expanding of the application scope to more tumors still need intensive research efforts. The clinical effects of T cellbased therapy have not been satisfactory; most tumor vaccinebased therapies are in phase Ⅰ and Ⅱ clinical trial, and the outcomes of few phase Ⅲ clinical trials are not satisfactory, leaving more work to be done for improvement. As we understand more about the roles of antibody in immune surveillance, it will help to make immunotherapy of tumors a promising strategy.
New tumor antigens are continuously being discovered due to the improvement of immunologic techniques. Whether dendritic cells induce immune activation or immune inhibition after they capture tumor antigens depends on the danger signals (GMCSF, MCP1, and HSP) or the inhibitory signals (TGFβ, IDO, and iNOS) released by tumor cells. Under the regulation of danger signals, dendritic cells activate Th1 immune response and eliminate tumors; under the regulation of inhibitory signals, they activate Th2 immune response and can not effectively eliminate tumors. Progress in tumor immunotherapy mainly is manifested by antibodybased therapy, T cellbased therapy, and tumor vaccinebased therapy. To day at least 7 antibodies have been confirmed effective when combined with chemotherapeutic agents in treatment of tumors. Despite of the progress made in antibody therapy, discovery of new targets, development of new antibodies, and expanding of the application scope to more tumors still need intensive research efforts. The clinical effects of T cellbased therapy have not been satisfactory; most tumor vaccinebased therapies are in phase Ⅰ and Ⅱ clinical trial, and the outcomes of few phase Ⅲ clinical trials are not satisfactory, leaving more work to be done for improvement. As we understand more about the roles of antibody in immune surveillance, it will help to make immunotherapy of tumors a promising strategy.
2009, 16(5):431-435.
DOI: 10.3872/j.issn.1007-385X.2009.5.002
Abstract:
Objective:To observe the inhibitory effect of CD4+CD25+CCR6+ regulatory T cells (CCR6+ Tregs) against CD8+T cells in vivo, and to investigate its relationship with tumor immune escape. Methods: Mouse mammary carcinoma models were established by inoculating mammary carcinoma 4T1 cells into nude mice. CCR6+ Tregs were isolated by FACS, and the Foxp3 expression on CCR6+ Tregs was further analyzed by FACS. 4T1 specific CD8+T cells were labeled with CFSE after isolation by FACS, and then transferred into 4T1 bearing nude mice combined with or without CCR6+ Tregs or CD4+CD25+CCR6regulatory T cells (CCR6 Tregs). Tumor growth and survival of 4T1 bearing mice were observed. The proliferation, IFNγ production, and granzyme B expression of CD8+T cells were examined by FACS. Results: Both CCR6+ Tregs and CCR6 Tregs expressed high levels of Foxp3. The tumors in CCR6+ Tregs and CD8+T cells cotransferred mice grew faster than those in CCR6 Tregs cotransferred and CD8+T celltransferred groups. The survival period of 4T1 bearing mice was significantly decreased in CCR6+ Tregs cotransferred group (P<0.05). Furthermore, the proliferation, IFNγ production and granzyme B expression of CD8+ T cells were also dramatically decreased in CCR6+ Tregs cotransferred group compared with those in CCR6 Tregs cotransferred and CD8+ T celltransferred groups (P<0.05). Conclusion: CCR6+ Tregs can effectively inhibit the function of CD8+ T cells, which might play an important role in tumor immune escape, tumor development and progress.
Objective:To observe the inhibitory effect of CD4+CD25+CCR6+ regulatory T cells (CCR6+ Tregs) against CD8+T cells in vivo, and to investigate its relationship with tumor immune escape. Methods: Mouse mammary carcinoma models were established by inoculating mammary carcinoma 4T1 cells into nude mice. CCR6+ Tregs were isolated by FACS, and the Foxp3 expression on CCR6+ Tregs was further analyzed by FACS. 4T1 specific CD8+T cells were labeled with CFSE after isolation by FACS, and then transferred into 4T1 bearing nude mice combined with or without CCR6+ Tregs or CD4+CD25+CCR6regulatory T cells (CCR6 Tregs). Tumor growth and survival of 4T1 bearing mice were observed. The proliferation, IFNγ production, and granzyme B expression of CD8+T cells were examined by FACS. Results: Both CCR6+ Tregs and CCR6 Tregs expressed high levels of Foxp3. The tumors in CCR6+ Tregs and CD8+T cells cotransferred mice grew faster than those in CCR6 Tregs cotransferred and CD8+T celltransferred groups. The survival period of 4T1 bearing mice was significantly decreased in CCR6+ Tregs cotransferred group (P<0.05). Furthermore, the proliferation, IFNγ production and granzyme B expression of CD8+ T cells were also dramatically decreased in CCR6+ Tregs cotransferred group compared with those in CCR6 Tregs cotransferred and CD8+ T celltransferred groups (P<0.05). Conclusion: CCR6+ Tregs can effectively inhibit the function of CD8+ T cells, which might play an important role in tumor immune escape, tumor development and progress.
3 Isolation, cultivation and identification of cancer stemlike cells from human liver cancer tissues
2009, 16(5):436-441.
DOI: 10.3872/j.issn.1007-385X.2009.5.003
Abstract:
Objective:To isolate and cultivate liver cancer stemlike cells (hLCSLCs) from human liver cancer specimens, so as to lay a foundation for stem celltargeted therapy of liver cancer. Methods: The hLCSLCs were obtained from fresh liver cancer tissue using enzymatic digestion and shortterm primary culture. Serumfree medium suitable for suspension sphere formation of hLCSLCs was selected by culturing them with serumfree medium supplement with heparin, albumin or hydrocortisone. Numbers of side population (SP) cells, and expressions of CD133 and CD90 in the hLCSLCs and sphere cells were examined by flow cytometry assay. Tumor formation ability of hLCSLCs and sphere cells were assessed by tumor formation assay. Results: hLCSLCs were successfully obtained from human liver cancer tissues, with SP cells being 0.9%, CD133 positive cells being 0.8% and CD90 positive cells being 12.7% in the hLCSLCs. The tumor formation rate was 100% when 2 000 SP cells were subcutaneously injected into nude mice. Serumfree medium containing heparin was more suitable for sphere formation of hLCSLCs, with SP cells being 4.6%, CD133+ cells being 9.7%, and CD90+ cells being 48% in these sphere cells. The tumor formation rate was 100% when 10 000 sphere cells were injected into nude mice. Conclusion: We have successfully established a method for isolating hLCSLCs from human liver cancer tissues. Serumfree suspension medium containing heparin can effectively enrich hLCSLCs .
Objective:To isolate and cultivate liver cancer stemlike cells (hLCSLCs) from human liver cancer specimens, so as to lay a foundation for stem celltargeted therapy of liver cancer. Methods: The hLCSLCs were obtained from fresh liver cancer tissue using enzymatic digestion and shortterm primary culture. Serumfree medium suitable for suspension sphere formation of hLCSLCs was selected by culturing them with serumfree medium supplement with heparin, albumin or hydrocortisone. Numbers of side population (SP) cells, and expressions of CD133 and CD90 in the hLCSLCs and sphere cells were examined by flow cytometry assay. Tumor formation ability of hLCSLCs and sphere cells were assessed by tumor formation assay. Results: hLCSLCs were successfully obtained from human liver cancer tissues, with SP cells being 0.9%, CD133 positive cells being 0.8% and CD90 positive cells being 12.7% in the hLCSLCs. The tumor formation rate was 100% when 2 000 SP cells were subcutaneously injected into nude mice. Serumfree medium containing heparin was more suitable for sphere formation of hLCSLCs, with SP cells being 4.6%, CD133+ cells being 9.7%, and CD90+ cells being 48% in these sphere cells. The tumor formation rate was 100% when 10 000 sphere cells were injected into nude mice. Conclusion: We have successfully established a method for isolating hLCSLCs from human liver cancer tissues. Serumfree suspension medium containing heparin can effectively enrich hLCSLCs .
MA Jichun
,
TAI Guixiang
,
ZHAO Xiaoxia
,
FANG Fang
,
ZHANG Qingyong
,
DOU Rui
,
CHEN Wenbo
,
LIU Zhonghui
2009, 16(5):442-446.
DOI: 10.3872/j.issn.1007-385X.2009.5.004
Abstract:
Objective:To investigate the inhibitory mechanism of mucin 1 tandem repeats peptide (MUC1 peptide) against the proliferation of tumor cells. Methods: Jurkat, Raji, U937, MCF7, SMMC7721, activated T and RAW264.7 cells were cocultured with MUC1 peptide; the inhibitory effects of MUC1 peptide on these cells were observed. Jurkat cellinoculated BABL/c mouse model was established by s.c. injection of Jurkat cells, and then the tumorbearing mice were treated with MUC1 peptide. The protein interacting with MUC1 peptide was identified by GST pulldown assay. Results: MUC1 peptide inhibited the proliferation of Jurkat, Raji, U937, MCF7 and SMMC7721 cells, but had no measurable inhibitory effect on activated T cells and RAW264.7 cells. MUC1 peptide significantly inhibited the growth of implanted Jurkat tumors in BABL/c mice (P<0.05). The protein interacting with MUC1 peptide in the lysates of Jurkat and MCF7 cells was confirmed by GST pulldown assay, with its molecular weight being approximately 115 000. The protein could bind specifically to antiMUC1 tandem repeat antibodies (GP1.4 and HMPV) and antiMUC1 cytoplasmic domain antibody (Ab5), indicating it might be a novel isotype of MUC1, and it was named small MUC1 (sMUC1). Conclusion: MUC1 peptide can transduce growth inhibitory signal by interacting with small MUC1 on the surface of tumor cells.
Objective:To investigate the inhibitory mechanism of mucin 1 tandem repeats peptide (MUC1 peptide) against the proliferation of tumor cells. Methods: Jurkat, Raji, U937, MCF7, SMMC7721, activated T and RAW264.7 cells were cocultured with MUC1 peptide; the inhibitory effects of MUC1 peptide on these cells were observed. Jurkat cellinoculated BABL/c mouse model was established by s.c. injection of Jurkat cells, and then the tumorbearing mice were treated with MUC1 peptide. The protein interacting with MUC1 peptide was identified by GST pulldown assay. Results: MUC1 peptide inhibited the proliferation of Jurkat, Raji, U937, MCF7 and SMMC7721 cells, but had no measurable inhibitory effect on activated T cells and RAW264.7 cells. MUC1 peptide significantly inhibited the growth of implanted Jurkat tumors in BABL/c mice (P<0.05). The protein interacting with MUC1 peptide in the lysates of Jurkat and MCF7 cells was confirmed by GST pulldown assay, with its molecular weight being approximately 115 000. The protein could bind specifically to antiMUC1 tandem repeat antibodies (GP1.4 and HMPV) and antiMUC1 cytoplasmic domain antibody (Ab5), indicating it might be a novel isotype of MUC1, and it was named small MUC1 (sMUC1). Conclusion: MUC1 peptide can transduce growth inhibitory signal by interacting with small MUC1 on the surface of tumor cells.
YANG Ming
,
FAN Dongmei
,
GAO Yingdai
,
ZHAO Yingxin
,
ZHOU Yuan
,
XU Yuanfu
,
JI Qing
,
WANG Jinhong
,
XIONG Dongsheng
,
YANG Chunzheng
2009, 16(5):447-451.
DOI: 10.3872/j.issn.1007-385X.2009.5.005
Abstract:
Objective:To observe the effects of cytarabine (AraC) on B7 expression on leukemia cells, and to study the effects of antiCD3/antiPgp bispecific antibody on the cytotoxicity of T cells against drugresistant leukemia cells. Methods：The expressions of B71 and B72 on K562 (leukemia cells) and K562/A02 cells (drugresistant leukemia cells) were examined by flow cytometry after treatment with AraC for different periods. B71 and B72 mRNA expressions in K562 and K562/A02 cells were detected by RTPCR. The proliferation of T lymphocytes stimulated by AraCtreated K562 and K562/A02 cells was detected by MTT assay. In vitro cytotoxicity of T lymphocytes against K562 and K562/A02 cells was analyzed using CytoTox 96 nonradioactive method after treatment with antiCD3/antiPgp bispecific antibody and AraC. Results:Compared with untreated cells, B71 and B72 expression on AraCtreated K562 and K562/A02 cells was significantly enhanced. MTT results showed that AraCtreated K562 and K562/A02 cells increased the proliferation of T lymhocytes. AraC combined with antiCD3/antiPgp bispecific antibody enhanced the cytotoxicity of T cells against K562 and K562/A02 target cells (T∶target, 0.39∶125∶1), especially when against Pgp positive drugresistant K562/A02 leukemia cells. Conclusion: AraC can upregulate B7 expression on leukemia cells, and when combined with antiCD3/antiPgp bispecific antibody it can enhance the cytotoxicity of T cells against target leukemia cells in vitro.
Objective:To observe the effects of cytarabine (AraC) on B7 expression on leukemia cells, and to study the effects of antiCD3/antiPgp bispecific antibody on the cytotoxicity of T cells against drugresistant leukemia cells. Methods：The expressions of B71 and B72 on K562 (leukemia cells) and K562/A02 cells (drugresistant leukemia cells) were examined by flow cytometry after treatment with AraC for different periods. B71 and B72 mRNA expressions in K562 and K562/A02 cells were detected by RTPCR. The proliferation of T lymphocytes stimulated by AraCtreated K562 and K562/A02 cells was detected by MTT assay. In vitro cytotoxicity of T lymphocytes against K562 and K562/A02 cells was analyzed using CytoTox 96 nonradioactive method after treatment with antiCD3/antiPgp bispecific antibody and AraC. Results:Compared with untreated cells, B71 and B72 expression on AraCtreated K562 and K562/A02 cells was significantly enhanced. MTT results showed that AraCtreated K562 and K562/A02 cells increased the proliferation of T lymhocytes. AraC combined with antiCD3/antiPgp bispecific antibody enhanced the cytotoxicity of T cells against K562 and K562/A02 target cells (T∶target, 0.39∶125∶1), especially when against Pgp positive drugresistant K562/A02 leukemia cells. Conclusion: AraC can upregulate B7 expression on leukemia cells, and when combined with antiCD3/antiPgp bispecific antibody it can enhance the cytotoxicity of T cells against target leukemia cells in vitro.
2009, 16(5):452-457.
DOI: 10.3872/j.issn.1007-385X.2009.5.006
Abstract:
Objective:To construct an xenoantigen synthetase α1,3 galactosyltransferase (α1,3GT) eukaryotic expression vector regulated by human telomerase catalytic subunit (hTERT) promoter, and to investigate its targeting expression of α1,3GT in lung cancer cell lines. Methods: Previously prepared and confirmed pig α1,3GT gene was inserted into pEGFPhTERTp plasmid to construct eukaryotic expression vector pEGFPhTERTpGT. pEGFPhTERTpGT and pEGFPN1GT (α1,3GT eukaryotic expression vector under the control of CMV promoter) were transfected into telomerasepositive human lung adenocarcinoma A549 cells and telomerasenegative human embryonic lung fibroblast MRC5 cells. 1,3GT mRNA expression in the transfected cells was detected by RTPCR. Expression of αgal antigen in transfected cells was examined by immunofluorescence and flow cytometry. Results: pEGFPhTERTpGT eukaryotic expression vector was successfully constructed. Both A549 and MRC5 cells transfected with pEGFPN1GT showed expression of α1,3GTmRNA; A549 cells but not telomerasenegative MRC5 cells expressed α1,3GT mRNA after transfection with pEGFPhTERTpGT. Furthermore, both A549 and MRC5 cells transfected with pEGFPN1GT showed expression of xenoantigen αgal; A549 but not MRC5 cells expressed xenoantigen αgal after transfection with pEGFPhTERTpGT (P<0.01). Conclusion: α1,3GT gene under the regulation of hTERT promoter can be specifically expressed in telomerasepositive lung cancer cell lines, which can induce production of xenoantigen αgal.
Objective:To construct an xenoantigen synthetase α1,3 galactosyltransferase (α1,3GT) eukaryotic expression vector regulated by human telomerase catalytic subunit (hTERT) promoter, and to investigate its targeting expression of α1,3GT in lung cancer cell lines. Methods: Previously prepared and confirmed pig α1,3GT gene was inserted into pEGFPhTERTp plasmid to construct eukaryotic expression vector pEGFPhTERTpGT. pEGFPhTERTpGT and pEGFPN1GT (α1,3GT eukaryotic expression vector under the control of CMV promoter) were transfected into telomerasepositive human lung adenocarcinoma A549 cells and telomerasenegative human embryonic lung fibroblast MRC5 cells. 1,3GT mRNA expression in the transfected cells was detected by RTPCR. Expression of αgal antigen in transfected cells was examined by immunofluorescence and flow cytometry. Results: pEGFPhTERTpGT eukaryotic expression vector was successfully constructed. Both A549 and MRC5 cells transfected with pEGFPN1GT showed expression of α1,3GTmRNA; A549 cells but not telomerasenegative MRC5 cells expressed α1,3GT mRNA after transfection with pEGFPhTERTpGT. Furthermore, both A549 and MRC5 cells transfected with pEGFPN1GT showed expression of xenoantigen αgal; A549 but not MRC5 cells expressed xenoantigen αgal after transfection with pEGFPhTERTpGT (P<0.01). Conclusion: α1,3GT gene under the regulation of hTERT promoter can be specifically expressed in telomerasepositive lung cancer cell lines, which can induce production of xenoantigen αgal.
2009, 16(5):458-463.
DOI: 10.3872/j.issn.1007-385X.2009.5.007
Abstract:
Objective:To prepare TGFβ insensitive cytotoxic T lymphocytes（CTLs）against prostate cancer and to observe its therapeutic effect against prostate cancer. Methods: Peripheral blood mononuclear cells were obtained from prostate cancer patients, and were induced to differentiate into dendritic cells in vitro. Prostate cancer antigen was prepared from the same patients, and tumor antigen specific CTLs were induced by prostate tumor antigen impulsedDCs. The tumor antigen specific CTLs were infected with lentivirus containing TβRⅡDNglytk gene, and the response of TβRⅡDNglytk transfectedCTLs to TGFβ was examined by Western blotting analysis. Prostate cancer cellinoculated mice were treated with TGFβ sensitive and insensitiveCTLs separately, and the therapeutic results were compared. Results: CTLs induced by prostate tumor antigen impulsedDCs grew significantly faster than those induced by nonimpulsedDCs (P<001). TβRⅡDNglytk gene infection reduced the sensitivity of CTLs to TGFβ. The reaction of CTLs infected by lentiviral vector to TGFβ was significantly weaker than that of noninfected CTLs. TGFβ insensitive CTLs significantly inhibited the growth of implanted prostate tumors in mice. Conclusion: TGFβ insensitive prostate tumor antigen specific CTLs have apparent therapeutic effect against prostate cancer, which paves a way for immunotherapy of prostate cancer.
Objective:To prepare TGFβ insensitive cytotoxic T lymphocytes（CTLs）against prostate cancer and to observe its therapeutic effect against prostate cancer. Methods: Peripheral blood mononuclear cells were obtained from prostate cancer patients, and were induced to differentiate into dendritic cells in vitro. Prostate cancer antigen was prepared from the same patients, and tumor antigen specific CTLs were induced by prostate tumor antigen impulsedDCs. The tumor antigen specific CTLs were infected with lentivirus containing TβRⅡDNglytk gene, and the response of TβRⅡDNglytk transfectedCTLs to TGFβ was examined by Western blotting analysis. Prostate cancer cellinoculated mice were treated with TGFβ sensitive and insensitiveCTLs separately, and the therapeutic results were compared. Results: CTLs induced by prostate tumor antigen impulsedDCs grew significantly faster than those induced by nonimpulsedDCs (P<001). TβRⅡDNglytk gene infection reduced the sensitivity of CTLs to TGFβ. The reaction of CTLs infected by lentiviral vector to TGFβ was significantly weaker than that of noninfected CTLs. TGFβ insensitive CTLs significantly inhibited the growth of implanted prostate tumors in mice. Conclusion: TGFβ insensitive prostate tumor antigen specific CTLs have apparent therapeutic effect against prostate cancer, which paves a way for immunotherapy of prostate cancer.
2009, 16(5):464-468.
DOI: 10.3872/j.issn.1007-385X.2009.5.008
Abstract:
Objective:To express hPDL1Ig fusion protein in the Pichia pastoris expression system, and to investigate the biological activity of the protein. Methods: The hPDL1IgG4 fusion gene was chemically synthesized and cloned into Pichia yeast expression vector. The recombinant vector was then transfected into GS115 Pichia yeast strain, and the fusion protein was purified by affinity chromatograph and ion exchange method before further identified by SDSPAGE and Western blotting. The binding ability of hPDL1Ig fusion protein to PD1 receptor was examined by ELISA. The inhibitory effects of hPDL1Ig fusion protein on T cells and on cytotoxicity of CTL against colon cancer SW480 cells were examined by MLR and 51Cr release assay, respectively. Results: The expression vector pPIC9KPDL1IgG4 was successfully constructed, and hPDL1IgG4 fusion protein was secreted by GS115 Pichia yeast strain, with the molecular weight being about 55 000 and the concentration being 120 μg/ml. The fusion protein was purified using fermentation strain. PDL1Ig fusion protein had high affinity with PD1 receptor; it also significantly inhibited the proliferation and activation of T cells (P<0.01) and the cytotoxicity of CTL against colon tumor cells (P<0.01). Conclusion: The hPDL1IgG4 fusion protein has been successfully prepared, and it has active biological functions, which lays a foundation for further investigating its regulatory effect on tumor immune response.
Objective:To express hPDL1Ig fusion protein in the Pichia pastoris expression system, and to investigate the biological activity of the protein. Methods: The hPDL1IgG4 fusion gene was chemically synthesized and cloned into Pichia yeast expression vector. The recombinant vector was then transfected into GS115 Pichia yeast strain, and the fusion protein was purified by affinity chromatograph and ion exchange method before further identified by SDSPAGE and Western blotting. The binding ability of hPDL1Ig fusion protein to PD1 receptor was examined by ELISA. The inhibitory effects of hPDL1Ig fusion protein on T cells and on cytotoxicity of CTL against colon cancer SW480 cells were examined by MLR and 51Cr release assay, respectively. Results: The expression vector pPIC9KPDL1IgG4 was successfully constructed, and hPDL1IgG4 fusion protein was secreted by GS115 Pichia yeast strain, with the molecular weight being about 55 000 and the concentration being 120 μg/ml. The fusion protein was purified using fermentation strain. PDL1Ig fusion protein had high affinity with PD1 receptor; it also significantly inhibited the proliferation and activation of T cells (P<0.01) and the cytotoxicity of CTL against colon tumor cells (P<0.01). Conclusion: The hPDL1IgG4 fusion protein has been successfully prepared, and it has active biological functions, which lays a foundation for further investigating its regulatory effect on tumor immune response.
2009, 16(5):469-473.
DOI: 10.3872/j.issn.1007-385X.2009.5.009
Abstract:
Objective:To construct a lentiviral livin shRNA vector to silence livin gene expression, and to study its effect on apoptosis of lung carcinoma cells. Methods:Livin expression in human lung adenocarcina SPCA1 cells was silenced by lentiviral livin shRNA infection. The morphology of apoptotic cells was observed by propidine iodide staining and fluoroscope; apoptosis rate and sublipliod apoptotic peak of SPCA1 cells were assessed by flow cytometry; expression of livin and caspase 3 in SPCA1 cells was examined by realtime PCR and Western blotting analysis. Results: Livin was constitutively expressed in SPCA1 cells. After livin expression was silenced by lentiviral livin shRNA infection, SPCA1 cells showed the characteristic morphology of apoptosis under fluoroscope, and the sublipliod apoptotic peak was identified by flow cytometry. Apoptosis rate in livin shRNA infected SPCA1 cells was significantly higher than that in blank and negative control groups (8.3% vs 0.08% and 0.13%, P<0.05). caspase 3 mRNA expression in SPCA1 cells had no change but the expression of cleavedcaspase 3 was greatly upregulated after lentiviral livin shRNA infection as showed by RTPCR and Western blotting analysis. Conclusion: Lentiviral livin shRNA can inhibit livin expression in human lung adenocarcina SPCA1 cells and induce cell apopotosis.
Objective:To construct a lentiviral livin shRNA vector to silence livin gene expression, and to study its effect on apoptosis of lung carcinoma cells. Methods:Livin expression in human lung adenocarcina SPCA1 cells was silenced by lentiviral livin shRNA infection. The morphology of apoptotic cells was observed by propidine iodide staining and fluoroscope; apoptosis rate and sublipliod apoptotic peak of SPCA1 cells were assessed by flow cytometry; expression of livin and caspase 3 in SPCA1 cells was examined by realtime PCR and Western blotting analysis. Results: Livin was constitutively expressed in SPCA1 cells. After livin expression was silenced by lentiviral livin shRNA infection, SPCA1 cells showed the characteristic morphology of apoptosis under fluoroscope, and the sublipliod apoptotic peak was identified by flow cytometry. Apoptosis rate in livin shRNA infected SPCA1 cells was significantly higher than that in blank and negative control groups (8.3% vs 0.08% and 0.13%, P<0.05). caspase 3 mRNA expression in SPCA1 cells had no change but the expression of cleavedcaspase 3 was greatly upregulated after lentiviral livin shRNA infection as showed by RTPCR and Western blotting analysis. Conclusion: Lentiviral livin shRNA can inhibit livin expression in human lung adenocarcina SPCA1 cells and induce cell apopotosis.
2009, 16(5):474-478.
DOI: 10.3872/j.issn.1007-385X.2009.5.010
Abstract:
Objective:To examine the effects of troglitazone (TGZ), agonist of peroxisome proliferatoractivated receptorγ (PPARγ), on the growth and growth hormone (GH) secretion of pituitary adenoma cells, and to expolre the possible mechanism. Methods: The inhibitory effect of TGZ on the proliferation and GH secretion of rat pituitary adenoma cell line GH3 was detected by MTT assay and ELISA. Furthermore, apoptosis, cell cycle as well as caspase3, Bcl2 and Bax expression of GH3 cells were examined by transmission electron microscopy, flow cytometry and Western blotting, respectively. Results: TGZ dose and timedependently inhibited the proliferation and GH secretion of GH3 cells. GH3 cells treated with TGZ had a typical morphological characteristic of apoptosis. CH3 cell number in G1 phase was increased and cell number in G2, S phases was significantly decreased after treatment with TGZ. The expression of caspase3 and Bax in CH3 cells was significantly increased and Bcl2 expression was markedly decreased in a dosedependent manner after treatment with TGZ. Conclusion: PPARγ agonist inhibits the growth and GH secretion of pituitary adenoma cells through inducing apoptosis and cell cycle arresting.
Objective:To examine the effects of troglitazone (TGZ), agonist of peroxisome proliferatoractivated receptorγ (PPARγ), on the growth and growth hormone (GH) secretion of pituitary adenoma cells, and to expolre the possible mechanism. Methods: The inhibitory effect of TGZ on the proliferation and GH secretion of rat pituitary adenoma cell line GH3 was detected by MTT assay and ELISA. Furthermore, apoptosis, cell cycle as well as caspase3, Bcl2 and Bax expression of GH3 cells were examined by transmission electron microscopy, flow cytometry and Western blotting, respectively. Results: TGZ dose and timedependently inhibited the proliferation and GH secretion of GH3 cells. GH3 cells treated with TGZ had a typical morphological characteristic of apoptosis. CH3 cell number in G1 phase was increased and cell number in G2, S phases was significantly decreased after treatment with TGZ. The expression of caspase3 and Bax in CH3 cells was significantly increased and Bcl2 expression was markedly decreased in a dosedependent manner after treatment with TGZ. Conclusion: PPARγ agonist inhibits the growth and GH secretion of pituitary adenoma cells through inducing apoptosis and cell cycle arresting.
2009, 16(5):479-483.
DOI: 10.3872/j.issn.1007-385X.2009.5.011
Abstract:
Objective:To investigate the effect of XIAP (Xlinked inhibitor of apoptosis protein)targeting siRNA on the proliferation of LoVo colorectal cancer cells in vitro and in vivo. Methods: The XIAPtargeting siRNA pSil2.1shXIAP1 and pSil2.1shXIAP2 eukaryotic vectors were constructed and were transfected into LoVo cells using Lipofectamine; the stable transfectants LoVoshXIAP2 were selected by G418. The expressions of XIAP mRNA and protein were determined by RTPCR and Western blotting. The proliferation of LoVo cells was determined by MTT and colony formation assay. Cell apoptosis was examined by FCM. The influence of XIAP knockdown on the proliferation of LoVo cells in vivo was observed in LoVobearing nude mice. Results:The expressions of XIAP mRNA and protein in LoVoshXIAP2 cells were significantly downregulated in LoVoshXIAP2 cells. Compared with untransfected LoVo cells, the proliferation and colony formation abilities of LoVoshXIAP2 cells were significantly inhibited (P<0.05); the apoptosis rate of LoVoshXIAP2 cells was significantly increased (P<0.05). The expression of XIAP protein in LoVoshXIAP2 implanted tumors was downregulated and the growth of tumors was significantly inhibited (all P<0.05). Conclusion: pSil2.1shXIAP2 plasmid can downregulate the expression of XIAP in LoVo cells and inhibit proliferation of LoVo cells in vitro and in vivo, making XIAPtargeting siRNA a potential new agent in immune therapy of colorectal cancer.
Objective:To investigate the effect of XIAP (Xlinked inhibitor of apoptosis protein)targeting siRNA on the proliferation of LoVo colorectal cancer cells in vitro and in vivo. Methods: The XIAPtargeting siRNA pSil2.1shXIAP1 and pSil2.1shXIAP2 eukaryotic vectors were constructed and were transfected into LoVo cells using Lipofectamine; the stable transfectants LoVoshXIAP2 were selected by G418. The expressions of XIAP mRNA and protein were determined by RTPCR and Western blotting. The proliferation of LoVo cells was determined by MTT and colony formation assay. Cell apoptosis was examined by FCM. The influence of XIAP knockdown on the proliferation of LoVo cells in vivo was observed in LoVobearing nude mice. Results:The expressions of XIAP mRNA and protein in LoVoshXIAP2 cells were significantly downregulated in LoVoshXIAP2 cells. Compared with untransfected LoVo cells, the proliferation and colony formation abilities of LoVoshXIAP2 cells were significantly inhibited (P<0.05); the apoptosis rate of LoVoshXIAP2 cells was significantly increased (P<0.05). The expression of XIAP protein in LoVoshXIAP2 implanted tumors was downregulated and the growth of tumors was significantly inhibited (all P<0.05). Conclusion: pSil2.1shXIAP2 plasmid can downregulate the expression of XIAP in LoVo cells and inhibit proliferation of LoVo cells in vitro and in vivo, making XIAPtargeting siRNA a potential new agent in immune therapy of colorectal cancer.
2009, 16(5):484-489.
DOI: 10.3872/j.issn.1007-385X.2009.5.012
Abstract:
Objective:To explore the in vitro and in vivo inhibitory effects of deuteriumdepleted water (DDW) on the proliferation of human lung carcinoma cells, and to explore the possible mechanism. Methods: The inhibitory effect of DDW on the proliferation of human lung carcinoma A549 cells and human embryonic lung fibroblast HLF1 cells was examined by MTT assay; apoptosis of A549 cells was examined by TUNEL; and cell cycle was analyzed by flow cytometry. Mouse model of lung carcinoma was established by inoculating human lung carcinoma H460 cells into BALB/c nude mice, and the growth of implanted tumors was observed after DDW treatment for 60 d. Results: Compared with control group, A549 cells treated with 0.0025％, 0.0050％ or 0.0105％ DDW showed significantly decreased proliferation 10 h after treatment (P<0.01). Then the inhibitory effects of DDW gradually disappeared, but appeared 48 h later again, with the inhibitory effects at 72 h being significant (P<0.05). DDW at the same dosages showed no inhibition on the proliferation of HLF1 cells (P>0.05). TUNEL assay verified the apoptosis of DDWtreated A549 cells, and the apoptosis rate of DDWtreated A549 cells was significantly higher than that of control group（\[45.30±4.21\]% vs \[22.25±030\]%, P<0.01). Cells in S phase were significantly increased in DDWtreated A549 cells compared with those in the control group (P<0.05). The life quality of H460 cellinoculated nude mice treated with DDW was greatly improved, with the tumor inhibition rate being 30.08%. Conclusion: DDW can inhibit the proliferation of lung cancer cells within a certain range of dosage and in a fluctuating pattern; its mechanism might be associated with induction of apoptosis and cell cycle arrest in S phase of tumor cells.
Objective:To explore the in vitro and in vivo inhibitory effects of deuteriumdepleted water (DDW) on the proliferation of human lung carcinoma cells, and to explore the possible mechanism. Methods: The inhibitory effect of DDW on the proliferation of human lung carcinoma A549 cells and human embryonic lung fibroblast HLF1 cells was examined by MTT assay; apoptosis of A549 cells was examined by TUNEL; and cell cycle was analyzed by flow cytometry. Mouse model of lung carcinoma was established by inoculating human lung carcinoma H460 cells into BALB/c nude mice, and the growth of implanted tumors was observed after DDW treatment for 60 d. Results: Compared with control group, A549 cells treated with 0.0025％, 0.0050％ or 0.0105％ DDW showed significantly decreased proliferation 10 h after treatment (P<0.01). Then the inhibitory effects of DDW gradually disappeared, but appeared 48 h later again, with the inhibitory effects at 72 h being significant (P<0.05). DDW at the same dosages showed no inhibition on the proliferation of HLF1 cells (P>0.05). TUNEL assay verified the apoptosis of DDWtreated A549 cells, and the apoptosis rate of DDWtreated A549 cells was significantly higher than that of control group（\[45.30±4.21\]% vs \[22.25±030\]%, P<0.01). Cells in S phase were significantly increased in DDWtreated A549 cells compared with those in the control group (P<0.05). The life quality of H460 cellinoculated nude mice treated with DDW was greatly improved, with the tumor inhibition rate being 30.08%. Conclusion: DDW can inhibit the proliferation of lung cancer cells within a certain range of dosage and in a fluctuating pattern; its mechanism might be associated with induction of apoptosis and cell cycle arrest in S phase of tumor cells.
2009, 16(5):490-493.
DOI: 10.3872/j.issn.1007-385X.2009.5.013
Abstract:
Objective:To investigate the inhibitory effect of curcumine on the proliferation of breast cancer MCF7 cells and the related oxidative stress mechanism. Methods: Human breast cancer MCF7 cells were treated with different concentrations of curcumine (540 μmol/L), then the proliferation of MCF7 cells was examined by MTT assay after 6, 12, 24 and 48 h; the apoptosis rate of MCF7 cells was detected by flow cytometry (FCM); the levels of reactive oxygen species (ROS) in MCF7 cells were detected by DCFHDA staining; and the activities of superoxide dismutase (SOD) and catalase (CAT) in MCF7 cells were detected by xanthine oxidase assay and visible spectrophotometer, respectively. Results: Low level curcumine (5, 10 μmol/L) inhibited the proliferation of MCF7 cells (P<0.01) without inducing apoptosis; ROS levels in MCF7 cells were firstly increased and then declined gradually; meanwhile, SOD and CAT activities decreased initially and then gradually increased. High level curcumine (20, 40 μmol/L) significantly inhibited the proliferation of MCF7 cells and induced evident apoptosis (P<0.01), and ROS levels in MCF7 cells were elevated instantly and maintained for a long time after treatment; at the same time, SOD and CAT activities were significantly decreased (P<0.01). Conclusion: Curcumine has a double role of antioxidant and oxidation, and it functions in a dosedependent manner. Different concentrations of curcumine can inhibit the proliferation of tumor cells through regulating oxidative statuses in cells.
Objective:To investigate the inhibitory effect of curcumine on the proliferation of breast cancer MCF7 cells and the related oxidative stress mechanism. Methods: Human breast cancer MCF7 cells were treated with different concentrations of curcumine (540 μmol/L), then the proliferation of MCF7 cells was examined by MTT assay after 6, 12, 24 and 48 h; the apoptosis rate of MCF7 cells was detected by flow cytometry (FCM); the levels of reactive oxygen species (ROS) in MCF7 cells were detected by DCFHDA staining; and the activities of superoxide dismutase (SOD) and catalase (CAT) in MCF7 cells were detected by xanthine oxidase assay and visible spectrophotometer, respectively. Results: Low level curcumine (5, 10 μmol/L) inhibited the proliferation of MCF7 cells (P<0.01) without inducing apoptosis; ROS levels in MCF7 cells were firstly increased and then declined gradually; meanwhile, SOD and CAT activities decreased initially and then gradually increased. High level curcumine (20, 40 μmol/L) significantly inhibited the proliferation of MCF7 cells and induced evident apoptosis (P<0.01), and ROS levels in MCF7 cells were elevated instantly and maintained for a long time after treatment; at the same time, SOD and CAT activities were significantly decreased (P<0.01). Conclusion: Curcumine has a double role of antioxidant and oxidation, and it functions in a dosedependent manner. Different concentrations of curcumine can inhibit the proliferation of tumor cells through regulating oxidative statuses in cells.
2009, 16(5):494-497.
DOI: 10.3872/j.issn.1007-385X.2009.5.014
Abstract:
Objective:To investigate the inhibitory effect of Brucea javanica oil emulsion on cervical cancer SiHa cells in vitro and the related mechanisms. Methods: SiHa cells were treated with Brucea javanica oil emulsion（2.5, 5.0, 10, 20, 40, and 80 μg/ml）for 24, 48, and 72 h. The inhibitory effect of Brucea javanica oil emulsion on the proliferation of SiHa cells was examined by MTT; the ultrastructure of SiHa cells was observed by transmission electron microscope (TEM); the DNA fragment of SiHa cells was detected by agarose gel electrophoresis; and the apoptosis of SiHa cells was examined by flow cytometry through Annexin V and PI staining. Results:MTT results showed that Brucea javanica oil emulsion significantly inhibited the proliferation of SiHa cells in a dose and timedependent manner. The inhibitory rate of SiHa cells reached 92.43% after treatment with 80 μg/ml Brucea javanica oil emulsion for 72 h. Apoptotic bodies and a characteristic apoptosis DNA ladder of SiHa cells were observed after treatment with 20 μg/ml Brucea javanica oil emulsion. Flow cytometry results demonstrated that the apoptosis rates of SiHa cells were (8.02±241)%, (51.60±7.67)%, and (77.22±580)% 48 h after treatment with 10, 20, and 40 μg/ml Brucea javanica oil emulsion for 48 h, respectively. Conclusion: Brucea javanica oil emulsion can inhibit the proliferation of SiHa cells, probably by inducing apoptosis of SiHa cells.
Objective:To investigate the inhibitory effect of Brucea javanica oil emulsion on cervical cancer SiHa cells in vitro and the related mechanisms. Methods: SiHa cells were treated with Brucea javanica oil emulsion（2.5, 5.0, 10, 20, 40, and 80 μg/ml）for 24, 48, and 72 h. The inhibitory effect of Brucea javanica oil emulsion on the proliferation of SiHa cells was examined by MTT; the ultrastructure of SiHa cells was observed by transmission electron microscope (TEM); the DNA fragment of SiHa cells was detected by agarose gel electrophoresis; and the apoptosis of SiHa cells was examined by flow cytometry through Annexin V and PI staining. Results:MTT results showed that Brucea javanica oil emulsion significantly inhibited the proliferation of SiHa cells in a dose and timedependent manner. The inhibitory rate of SiHa cells reached 92.43% after treatment with 80 μg/ml Brucea javanica oil emulsion for 72 h. Apoptotic bodies and a characteristic apoptosis DNA ladder of SiHa cells were observed after treatment with 20 μg/ml Brucea javanica oil emulsion. Flow cytometry results demonstrated that the apoptosis rates of SiHa cells were (8.02±241)%, (51.60±7.67)%, and (77.22±580)% 48 h after treatment with 10, 20, and 40 μg/ml Brucea javanica oil emulsion for 48 h, respectively. Conclusion: Brucea javanica oil emulsion can inhibit the proliferation of SiHa cells, probably by inducing apoptosis of SiHa cells.
2009, 16(5):498-502.
DOI: 10.3872/j.issn.1007-385X.2009.5.015
Abstract:
Objective:To investigate the expression of glucoseregulated protein 78 in human esophageal squamous cell carcinoma and normal esophageal tissues, and to evaluate its correlation with clinical pathological characteristics of esophageal squamous cell carcinoma. Methods: Fiftynine specimens of human esophageal squamous cell carcinoma and twenty adjacent normal specimens were collected from the patients who received operation for esophageal squamous cell carcinomas in our Hospital between Oct. 2007 and Nov. 2008. GRP78 mRNA and GRP78 protein expressions were examined by RTPCR assay and Western blotting, respectively. The relationship between GRP78 expression with clinical parameters of patients, such as gender, length of tumor, tumor infiltration depth, tumor differentiation grade stage, and lymphatic metastasis, was analyzed. Results: GRP78 mRNA and GRP78 protein expression levels in human esophageal squamous cell carcinoma were significantly higher than those in the normal esophageal tissues (all P<0.01). GRP78 expression in esophageal squamous cell carcinoma tissues was correlated with the infiltration depth, grade of differentiation(P<0.05 or P<0.01), stage of tumors (P<0.01), and lymphatic metastasis (all P<0.01), but not with gender or the length of tumor (P>0.05). Conclusion: GRP78 participates in the development, progress of esophageal squamous cell carcinomas, and its expression is positively associated with the malignancy of carcinoma. GRP78 may be taken as a potential biomarker in evaluating the malignancy of esophageal squamous cell carcinoma.
Objective:To investigate the expression of glucoseregulated protein 78 in human esophageal squamous cell carcinoma and normal esophageal tissues, and to evaluate its correlation with clinical pathological characteristics of esophageal squamous cell carcinoma. Methods: Fiftynine specimens of human esophageal squamous cell carcinoma and twenty adjacent normal specimens were collected from the patients who received operation for esophageal squamous cell carcinomas in our Hospital between Oct. 2007 and Nov. 2008. GRP78 mRNA and GRP78 protein expressions were examined by RTPCR assay and Western blotting, respectively. The relationship between GRP78 expression with clinical parameters of patients, such as gender, length of tumor, tumor infiltration depth, tumor differentiation grade stage, and lymphatic metastasis, was analyzed. Results: GRP78 mRNA and GRP78 protein expression levels in human esophageal squamous cell carcinoma were significantly higher than those in the normal esophageal tissues (all P<0.01). GRP78 expression in esophageal squamous cell carcinoma tissues was correlated with the infiltration depth, grade of differentiation(P<0.05 or P<0.01), stage of tumors (P<0.01), and lymphatic metastasis (all P<0.01), but not with gender or the length of tumor (P>0.05). Conclusion: GRP78 participates in the development, progress of esophageal squamous cell carcinomas, and its expression is positively associated with the malignancy of carcinoma. GRP78 may be taken as a potential biomarker in evaluating the malignancy of esophageal squamous cell carcinoma.
2009, 16(5):503-506.
DOI: 10.3872/j.issn.1007-385X.2009.5.016
Abstract:
Objective:To investigate the expression of DNA polymerase iota (Polι) in human lung cancer tissues, adjacent lung cancer tissues, and normal lung tissues, so as to analyze the role of Polι gene in the development and progression of lung cancer. Methods: Samples were obtained from lung cancer patients who were surgically treated during Aug. 2003 to Jan. 2001 in Qilu Hospital of Shandong University. Immunohistochemisty and RTPCR were applied to detect Polι expression in 60 lung cancer specimens, 30 adjacent lung cancer specimens and 30 normal tissue specimens. Data were analyzed with PSS13.0 software. Results: The positive rates of Polι in lung cancer, adjacent lung cancer tissues and normal tissues were 54.8%, 33.3% and 10.0%, respectively. Polι expression levels were similar in different types of lung cancer (P>005). Polι expression in lung cancer tissues was decreased with the increase of differentiation grade of lung cancer (P<0.05), with the expression in grade Ⅲ and Ⅳ of lung cancer significantly higher than those in grade Ⅰ and Ⅱ(P<0.05). Polι expression in patients who survived less than 3 years was markedly higher than in those survived longer than 3 years (P<005); and smoking patients had a markedly higher expression than nonsmokers (P<0.05). Conclusion: High expression of Polι plays an important role in development and progression of lung cancer, and it may be used in evaluating the biological characteristics and prognosis of lung cancer.
Objective:To investigate the expression of DNA polymerase iota (Polι) in human lung cancer tissues, adjacent lung cancer tissues, and normal lung tissues, so as to analyze the role of Polι gene in the development and progression of lung cancer. Methods: Samples were obtained from lung cancer patients who were surgically treated during Aug. 2003 to Jan. 2001 in Qilu Hospital of Shandong University. Immunohistochemisty and RTPCR were applied to detect Polι expression in 60 lung cancer specimens, 30 adjacent lung cancer specimens and 30 normal tissue specimens. Data were analyzed with PSS13.0 software. Results: The positive rates of Polι in lung cancer, adjacent lung cancer tissues and normal tissues were 54.8%, 33.3% and 10.0%, respectively. Polι expression levels were similar in different types of lung cancer (P>005). Polι expression in lung cancer tissues was decreased with the increase of differentiation grade of lung cancer (P<0.05), with the expression in grade Ⅲ and Ⅳ of lung cancer significantly higher than those in grade Ⅰ and Ⅱ(P<0.05). Polι expression in patients who survived less than 3 years was markedly higher than in those survived longer than 3 years (P<005); and smoking patients had a markedly higher expression than nonsmokers (P<0.05). Conclusion: High expression of Polι plays an important role in development and progression of lung cancer, and it may be used in evaluating the biological characteristics and prognosis of lung cancer.
2009, 16(5):507-511.
DOI: 10.3872/j.issn.1007-385X.2009.5.017
Abstract:
Objective:To examine JAK2V617F mutation in BCRABL negative patients with myeloproliferative disorders (MPD) and its relationship with clinical characteristics of MPD.Methods: Fiftysix BCRABL negative MPD patients (who had been diagnosed in the Provincial Hospital Affiliated to Shandong University) were included in the present study. The patients included 20 with polycythaemia vera (PV), 26 with essential thrombocythaemia (ET) and 10 with idiopathic myelofibrosis (IMF). JAK2V617F mutation in MPD patients was detected by allelespecific polymerase chain reaction (ASPCR) and DNAsequencing, and its correlation with clinical characteristics of MPD was analyzed. Results: JAK2 V617F mutation was detected in 36 of the 56 BCRABL negative MPD patients, including 17 (17/20, 85%) with PV, 14 (14/26, 53.8%) with ET, and 5 (5/10, 50%) with IMF. The leukocyte (P=0.018) and platelet counts (P=0.021) were significantly different between JAK2V617 positive and negative MPD patients in PV group; the leukocyte counts (P=0.001) and hemoglobin (P=0.007) were significantly different in ET group; and the leukocyte counts were significantly different (P=0026) in IMF group. Significant difference was also found in the incidences of bleeding, thrombosis between JAK2V617 positive and negative patients in ET group(P=0.016), but not in PV or IMF group. Conclusion: ASPCR is a sensitive and reliable technique in detecting JAK2 V617F mutation. The clinical characteristics of JAK2 V617F mutantion positive MPD patients are different from those without the mutation.
Objective:To examine JAK2V617F mutation in BCRABL negative patients with myeloproliferative disorders (MPD) and its relationship with clinical characteristics of MPD.Methods: Fiftysix BCRABL negative MPD patients (who had been diagnosed in the Provincial Hospital Affiliated to Shandong University) were included in the present study. The patients included 20 with polycythaemia vera (PV), 26 with essential thrombocythaemia (ET) and 10 with idiopathic myelofibrosis (IMF). JAK2V617F mutation in MPD patients was detected by allelespecific polymerase chain reaction (ASPCR) and DNAsequencing, and its correlation with clinical characteristics of MPD was analyzed. Results: JAK2 V617F mutation was detected in 36 of the 56 BCRABL negative MPD patients, including 17 (17/20, 85%) with PV, 14 (14/26, 53.8%) with ET, and 5 (5/10, 50%) with IMF. The leukocyte (P=0.018) and platelet counts (P=0.021) were significantly different between JAK2V617 positive and negative MPD patients in PV group; the leukocyte counts (P=0.001) and hemoglobin (P=0.007) were significantly different in ET group; and the leukocyte counts were significantly different (P=0026) in IMF group. Significant difference was also found in the incidences of bleeding, thrombosis between JAK2V617 positive and negative patients in ET group(P=0.016), but not in PV or IMF group. Conclusion: ASPCR is a sensitive and reliable technique in detecting JAK2 V617F mutation. The clinical characteristics of JAK2 V617F mutantion positive MPD patients are different from those without the mutation.
WANG Liangzhe
,
LIU Huimin
,
YANG Zhihui
,
HE Jin
,
CAO Xu
,
LIU Fangfang
,
SUN Jing
,
ZHU Weijian
,
CHEN Bing
2009, 16(5):512-515.
DOI: 10.3872/j.issn.1007-385X.2009.5.018
Abstract:
Objective: To investigate the expression of osteopontin (OPN) in spinal plasma cell myeloma (PCM) and its relationship with the clinicopathological characteristics and prognoses of PCM patients. Methods: Fortyone PCM and 14 spinal simple bone cysts samples were obtained from Affiliated Changzheng Hospital of Second Military University (19982006) after surgery. The expression of OPN was determined by streptavidinperosidase (SP) immunohistochemical staining. Results: (1) The positive expression rate of OPN in spinal PCM tissues was 82.93% (34/41), and it was absent in spinal simple bone cyst tissues(P<0.01). (2) The expression of OPN in spinal PCM tissues was correlated with clinical and Mcomponent classifications of PCM (P<0.05)，but was not correlated with the type of light chain, pathological stage, age or gender of patients (P>0.05). (3) The overall survival rate of OPN positive PCM patients was higher than that of OPN negative patients (P<0.01). Conclusion: OPN negative PCM patients are liable to have a poor prognosis. The clinical and Mcomponent classifications combined with detection of OPN expression in PCM patients maybe used for predication of prognoses of PCM patients.
Objective: To investigate the expression of osteopontin (OPN) in spinal plasma cell myeloma (PCM) and its relationship with the clinicopathological characteristics and prognoses of PCM patients. Methods: Fortyone PCM and 14 spinal simple bone cysts samples were obtained from Affiliated Changzheng Hospital of Second Military University (19982006) after surgery. The expression of OPN was determined by streptavidinperosidase (SP) immunohistochemical staining. Results: (1) The positive expression rate of OPN in spinal PCM tissues was 82.93% (34/41), and it was absent in spinal simple bone cyst tissues(P<0.01). (2) The expression of OPN in spinal PCM tissues was correlated with clinical and Mcomponent classifications of PCM (P<0.05)，but was not correlated with the type of light chain, pathological stage, age or gender of patients (P>0.05). (3) The overall survival rate of OPN positive PCM patients was higher than that of OPN negative patients (P<0.01). Conclusion: OPN negative PCM patients are liable to have a poor prognosis. The clinical and Mcomponent classifications combined with detection of OPN expression in PCM patients maybe used for predication of prognoses of PCM patients.
ZHANG Jianjian
,
CHEN Hanping
,
PENG Xiang
,
WENG Ruiguang
,
LIU Jun
,
YE Hui
,
ZHU Yuan
,
ZHENG Weiming
,
LI Jianmin
2009, 16(5):516-519.
DOI: 10.3872/j.issn.1007-385X.2009.5.019
Abstract:
Objective:To examine the expression of IL13Rα2 and proliferating cell nuclear antigen (PCNA) in malignant glioma, and to investigate its relationship with clinical pathological characteristics and prognosis. Methods:Fortythree surgical samples of malignant glioma were obtained from First Affiliated Hospital of Wenzhou Medical College during 20022006. Expression of IL13Rα2 and PCNA proteins was examined by immunohistochemistry staining, and the integrated optical density (IOD) was analyzed by image analysis system. The correlation between the expression of IL13Rα2 with that of PCNA, and their relationship with clinical characteristics and prognosis of glioma were studied.Results: (1) The positive rates of IL13Rα2 and PCNA in malignant glioma tissues were 93% (40/43) and 84% (36/43), respectively. (2) No relationship of IL13Rα2 and PCNA expression with the location and tumor size of glioma was found (P>0.05). The expression of IL13Rα2 and PCNA in glioma was significantly different between Ⅲ grade and Ⅳ grade glioma tissues (P=0.031, P=0.002). Patients who survived less than 6 months had a significantly higher expression of IL13Rα2 than those who survived more than 6 months (P=0.028). (3) The expression of IL13Rα2 was positively correlated with that of PCNA (r=0.653, P=0.000). Conclusion: IL13Rα2 and PCNA are overexpressed in malignant glioma tissues, and IL13Rα2 expression is correlated with differentiation grade and prognosis of glioma. IL13Rα2 has potential roles in clinical diagnosis and prognositic evaluation of malignant glioma.
Objective:To examine the expression of IL13Rα2 and proliferating cell nuclear antigen (PCNA) in malignant glioma, and to investigate its relationship with clinical pathological characteristics and prognosis. Methods:Fortythree surgical samples of malignant glioma were obtained from First Affiliated Hospital of Wenzhou Medical College during 20022006. Expression of IL13Rα2 and PCNA proteins was examined by immunohistochemistry staining, and the integrated optical density (IOD) was analyzed by image analysis system. The correlation between the expression of IL13Rα2 with that of PCNA, and their relationship with clinical characteristics and prognosis of glioma were studied.Results: (1) The positive rates of IL13Rα2 and PCNA in malignant glioma tissues were 93% (40/43) and 84% (36/43), respectively. (2) No relationship of IL13Rα2 and PCNA expression with the location and tumor size of glioma was found (P>0.05). The expression of IL13Rα2 and PCNA in glioma was significantly different between Ⅲ grade and Ⅳ grade glioma tissues (P=0.031, P=0.002). Patients who survived less than 6 months had a significantly higher expression of IL13Rα2 than those who survived more than 6 months (P=0.028). (3) The expression of IL13Rα2 was positively correlated with that of PCNA (r=0.653, P=0.000). Conclusion: IL13Rα2 and PCNA are overexpressed in malignant glioma tissues, and IL13Rα2 expression is correlated with differentiation grade and prognosis of glioma. IL13Rα2 has potential roles in clinical diagnosis and prognositic evaluation of malignant glioma.
CUI Qiu
,
GUO Jun
,
LIU Shu-bin
,
JIANG Wei-hao
,
LIU Yao-shen
,
FAN Hai-tao
,
WANG Lei
,
ZHANG Bing
,
LI Ding-feng
2009, 16(5):520-522.
DOI: 10.3872/j.issn.1007-385X.2009.5.020
Abstract:
目的：探讨骨肉瘤组织中O(6)甲基鸟嘌呤DNA甲基转移酶\[O(6)methylguanineDNA methyltransferase，MGMT\]的表达与顺铂（cisplatin，DDP）治疗后瘤组织坏死程度的关系。方法：选取2001年1月至2008年4月北京307医院骨科收治的骨肉瘤患者76例，全部患者均以标准的DDP方案治疗3个疗程以上，然后手术切除肿瘤。以免疫组化法检测顺铂治疗前活检标本中MGMT的表达,以HE染色观察手术切除骨肉瘤组织的坏死程度，分析MGMT表达和肿瘤坏死程度的关系。结果：本组患者骨肉瘤组织中MGMT蛋白的表达率为68% （52/76），其毛细胞血管扩张型、成骨细胞型、成软骨细胞型、成纤维细胞瘤型间的MGMT表达率无差异。MGMT阴性的肿瘤经DDP化疗后的组织坏死率高，反之肿瘤坏死率低（P<0.01）。结论：MGMT蛋白表达与DDP化疗后骨肉瘤组织的坏死程度呈负相关，该指标部分地反映肿瘤的耐药性和DDP的疗效。
目的：探讨骨肉瘤组织中O(6)甲基鸟嘌呤DNA甲基转移酶\[O(6)methylguanineDNA methyltransferase，MGMT\]的表达与顺铂（cisplatin，DDP）治疗后瘤组织坏死程度的关系。方法：选取2001年1月至2008年4月北京307医院骨科收治的骨肉瘤患者76例，全部患者均以标准的DDP方案治疗3个疗程以上，然后手术切除肿瘤。以免疫组化法检测顺铂治疗前活检标本中MGMT的表达,以HE染色观察手术切除骨肉瘤组织的坏死程度，分析MGMT表达和肿瘤坏死程度的关系。结果：本组患者骨肉瘤组织中MGMT蛋白的表达率为68% （52/76），其毛细胞血管扩张型、成骨细胞型、成软骨细胞型、成纤维细胞瘤型间的MGMT表达率无差异。MGMT阴性的肿瘤经DDP化疗后的组织坏死率高，反之肿瘤坏死率低（P<0.01）。结论：MGMT蛋白表达与DDP化疗后骨肉瘤组织的坏死程度呈负相关，该指标部分地反映肿瘤的耐药性和DDP的疗效。
2009, 16(5):523-525.
DOI: 10.3872/j.issn.1007-385X.2009.5.021
Abstract:
目的：观察香菇多糖联合化疗治疗晚期非小细胞肺癌（nonsmall cell lung cancer,NSCLC）的治疗效果。方法：113例Ⅲ～Ⅳ期经病理学或细胞学证实的NSCLC的初治或复治晚期非小细胞肺癌患者随机分为A、B两组，A组(57例)采用香菇多糖加TP方案（多西他赛+顺铂）化疗，B组(56例)采用单纯TP方案化疗。对患者疗效、不良反应及T淋巴细胞亚群水平进行评价。结果：A、B两组的治疗有效率分别为43.85%和37.50%(P>0.05); B组的Ⅱ～Ⅳ度白细胞减少及恶心呕吐反应发生率(分别为34和32例)明显高于A组(分别为20和17例)( P<0.05)。A组T淋巴细胞中CD3、CD4 、CD4/CD8及NK显著高于B组(P<0.05)。结论：香菇多糖联合化疗治疗晚期非小细胞肺癌与单纯化疗相比疗效相当，但不良反应轻、免疫功能明显改善。
目的：观察香菇多糖联合化疗治疗晚期非小细胞肺癌（nonsmall cell lung cancer,NSCLC）的治疗效果。方法：113例Ⅲ～Ⅳ期经病理学或细胞学证实的NSCLC的初治或复治晚期非小细胞肺癌患者随机分为A、B两组，A组(57例)采用香菇多糖加TP方案（多西他赛+顺铂）化疗，B组(56例)采用单纯TP方案化疗。对患者疗效、不良反应及T淋巴细胞亚群水平进行评价。结果：A、B两组的治疗有效率分别为43.85%和37.50%(P>0.05); B组的Ⅱ～Ⅳ度白细胞减少及恶心呕吐反应发生率(分别为34和32例)明显高于A组(分别为20和17例)( P<0.05)。A组T淋巴细胞中CD3、CD4 、CD4/CD8及NK显著高于B组(P<0.05)。结论：香菇多糖联合化疗治疗晚期非小细胞肺癌与单纯化疗相比疗效相当，但不良反应轻、免疫功能明显改善。
2009, 16(5):526-531.
DOI: 10.3872/j.issn.1007-385X.2009.5.022
Abstract:
Notch信号转导通路由一组在进化上高度保守的细胞膜配体、受体及下游分子组成。细胞间受体配体作用可激活Notch信号转导过程，从而直接调节基因转录，使细胞基因表达受相邻细胞调控，Notch信号在细胞分化、胚胎发育、组织自我更新过程中均发挥了重要的作用，许多病理过程（包括肿瘤）都有Notch信号参与。Notch信号多作为癌基因促进肿瘤生长，但在某些组织也可起到诱导细胞分化、抑制肿瘤增殖的作用。肿瘤干细胞中Notch信号的改变可能发挥了关键性作用。目前认为，Notch在肝癌中作为抑癌基因抑制肿瘤的生长，其机制初步被认为是Notch1使JNK活化、p53高表达以及Bcl2表达下调，从而诱导肝癌细胞凋亡，但尚待更加深入的研究。鉴于针对Notch信号通路的干预措施已经成为治疗肿瘤的新方式，该通路也有望成为肝癌的生物治疗新的靶点。
Notch信号转导通路由一组在进化上高度保守的细胞膜配体、受体及下游分子组成。细胞间受体配体作用可激活Notch信号转导过程，从而直接调节基因转录，使细胞基因表达受相邻细胞调控，Notch信号在细胞分化、胚胎发育、组织自我更新过程中均发挥了重要的作用，许多病理过程（包括肿瘤）都有Notch信号参与。Notch信号多作为癌基因促进肿瘤生长，但在某些组织也可起到诱导细胞分化、抑制肿瘤增殖的作用。肿瘤干细胞中Notch信号的改变可能发挥了关键性作用。目前认为，Notch在肝癌中作为抑癌基因抑制肿瘤的生长，其机制初步被认为是Notch1使JNK活化、p53高表达以及Bcl2表达下调，从而诱导肝癌细胞凋亡，但尚待更加深入的研究。鉴于针对Notch信号通路的干预措施已经成为治疗肿瘤的新方式，该通路也有望成为肝癌的生物治疗新的靶点。
2009, 16(5):532-536.
DOI: 10.3872/j.issn.1007-385X.2009.5.023
Abstract:
间充质干细胞（mesenchymal stem cells，MSCs）是存在于骨髓等组织中的一种具有高度自我更新能力和多向分化潜能的非造血干细胞，具有对创伤及肿瘤组织较为特异的趋向性。MSCs与肿瘤微环境之间存在复杂的交互作用。一方面，MSCs可直接作用于肿瘤细胞抑制其生长；也可作为抗原提呈细胞，激活肿瘤抗原特异性免疫应答；还可作为细胞载体，传递和表达多种抗肿瘤治疗因子，参与抗肿瘤药物的运输、免疫应答的激活以及新生血管的抑制。另一方面，MSCs强大的分化和增殖能力促使其参与肿瘤组织的构建；通过多种趋化因子作用引起肿瘤细胞表型变化，促进肿瘤恶性行为；MSCs具有类似“免疫豁免”效应，可对各主要类型的免疫细胞产生增殖和活化抑制作用，有助于肿瘤细胞逃逸；表达于MSCs表面的多种生长因子和细胞因子可协同促进肿瘤血管和淋巴管生成，参与肿瘤侵袭、转移。MSCs尚具有恶变潜能，在某些条件下可自发转化为致瘤干细胞。
间充质干细胞（mesenchymal stem cells，MSCs）是存在于骨髓等组织中的一种具有高度自我更新能力和多向分化潜能的非造血干细胞，具有对创伤及肿瘤组织较为特异的趋向性。MSCs与肿瘤微环境之间存在复杂的交互作用。一方面，MSCs可直接作用于肿瘤细胞抑制其生长；也可作为抗原提呈细胞，激活肿瘤抗原特异性免疫应答；还可作为细胞载体，传递和表达多种抗肿瘤治疗因子，参与抗肿瘤药物的运输、免疫应答的激活以及新生血管的抑制。另一方面，MSCs强大的分化和增殖能力促使其参与肿瘤组织的构建；通过多种趋化因子作用引起肿瘤细胞表型变化，促进肿瘤恶性行为；MSCs具有类似“免疫豁免”效应，可对各主要类型的免疫细胞产生增殖和活化抑制作用，有助于肿瘤细胞逃逸；表达于MSCs表面的多种生长因子和细胞因子可协同促进肿瘤血管和淋巴管生成，参与肿瘤侵袭、转移。MSCs尚具有恶变潜能，在某些条件下可自发转化为致瘤干细胞。
2009, 16(5):537-540.
DOI: 10.3872/j.issn.1007-385X.2009.5.024
Abstract:
肿瘤的生长和转移离不开新生血管的形成。Hexastatin是一种新型的肿瘤血管生成抑制剂，它来源于细胞外基质，是Ⅳ型胶原蛋白α6链的非胶原区NC1，相对分子质量为25 000。Hexastatin的分布具有明显的组织特异性。Hexastatin可以通过一种RGD非依赖方式与整合素分子αvβ3结合，调节内皮细胞的黏附和迁移，并且剂量依赖性地抑制内皮细胞的增殖，对抑制血管生成起到显著的作用。Hexastatin能抑制80%~90%的经bFGF刺激的鸡胚绒毛膜血管的生成，抑制90%的人脐静脉内皮细胞的迁移，Hexastatin还能有力地（大约60%）抑制CS1黑素瘤细胞的生长。在已建立的肿瘤小鼠模型中， Hexastatin能抑制小鼠肺癌移植瘤生长。在癌细胞浸润过程中，α6(Ⅳ)链经常会在癌细胞巢穴周围的基底膜区域中缺失，使得Hexastatin在癌组织中含量下调，从而成为诊断肿瘤浸润的重要指标。Hexastatin作为新型的肿瘤血管生成抑制剂在肿瘤的防治中具有重要的研究和应用前景。
肿瘤的生长和转移离不开新生血管的形成。Hexastatin是一种新型的肿瘤血管生成抑制剂，它来源于细胞外基质，是Ⅳ型胶原蛋白α6链的非胶原区NC1，相对分子质量为25 000。Hexastatin的分布具有明显的组织特异性。Hexastatin可以通过一种RGD非依赖方式与整合素分子αvβ3结合，调节内皮细胞的黏附和迁移，并且剂量依赖性地抑制内皮细胞的增殖，对抑制血管生成起到显著的作用。Hexastatin能抑制80%~90%的经bFGF刺激的鸡胚绒毛膜血管的生成，抑制90%的人脐静脉内皮细胞的迁移，Hexastatin还能有力地（大约60%）抑制CS1黑素瘤细胞的生长。在已建立的肿瘤小鼠模型中， Hexastatin能抑制小鼠肺癌移植瘤生长。在癌细胞浸润过程中，α6(Ⅳ)链经常会在癌细胞巢穴周围的基底膜区域中缺失，使得Hexastatin在癌组织中含量下调，从而成为诊断肿瘤浸润的重要指标。Hexastatin作为新型的肿瘤血管生成抑制剂在肿瘤的防治中具有重要的研究和应用前景。
2009, 16(5):541-545.
DOI: 10.3872/j.issn.1007-385X.2009.5.025
Abstract:
上皮细胞间质转化(epithelialmesenchymal transition，EMT)是具有极性的上皮细胞转换成为具有移行能力的间质细胞并获得侵袭和迁移能力的过程，它存在于人体多个生理和病理过程中。EMT与肿瘤细胞的侵袭和转移有着密切的关系。E钙黏蛋白（Ecadherin,Ecad）用于维持正常细胞间连接的稳定性，其表达水平与EMT的发生以及肿瘤的侵袭能力呈负相关，是EMT的关键分子。转录因子如锌指蛋白Snaill、SIP1等可下调Ecad的表达而促进EMT；Slug、Twist也可诱导EMT的发生。多种生长因子如HGF、TGFβ等可通过细胞内多个不同信号转导途径调控EMT的发生，促进肿瘤的转移。细胞外基质（ECM）对EMT的发生起着重要的作用，整合素介导的细胞与基质之间的黏附作用改变可引起EMT，基质金属蛋白酶（MMPs）通过影响ECM诱导EMT的发生。不同的MicroRNA如microRNA 10b或microRNA 200对EMT发生有促进或抑制作用。发生EMT的肿瘤细胞可获得某些类似于干细胞的能力如自我更新等，而胚胎干细胞在分化时伴有类似EMT过程的分子生物学事件。EMT的分子生物学过程、信号转导通路还有待更深入地研究完善。
上皮细胞间质转化(epithelialmesenchymal transition，EMT)是具有极性的上皮细胞转换成为具有移行能力的间质细胞并获得侵袭和迁移能力的过程，它存在于人体多个生理和病理过程中。EMT与肿瘤细胞的侵袭和转移有着密切的关系。E钙黏蛋白（Ecadherin,Ecad）用于维持正常细胞间连接的稳定性，其表达水平与EMT的发生以及肿瘤的侵袭能力呈负相关，是EMT的关键分子。转录因子如锌指蛋白Snaill、SIP1等可下调Ecad的表达而促进EMT；Slug、Twist也可诱导EMT的发生。多种生长因子如HGF、TGFβ等可通过细胞内多个不同信号转导途径调控EMT的发生，促进肿瘤的转移。细胞外基质（ECM）对EMT的发生起着重要的作用，整合素介导的细胞与基质之间的黏附作用改变可引起EMT，基质金属蛋白酶（MMPs）通过影响ECM诱导EMT的发生。不同的MicroRNA如microRNA 10b或microRNA 200对EMT发生有促进或抑制作用。发生EMT的肿瘤细胞可获得某些类似于干细胞的能力如自我更新等，而胚胎干细胞在分化时伴有类似EMT过程的分子生物学事件。EMT的分子生物学过程、信号转导通路还有待更深入地研究完善。
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.