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Volume 16,Issue 4,2009 Table of Contents
2009, 16(4):319-324.
DOI: 10.3872/j.issn.1007-385X.2009.4.001
Abstract:
Myeloidderived suppressor cells (MDSCs) are heterogeneous cells derived from myeloid progenitor cells and immature myeloid cells (IMCs) in bone marrow; they are the progenitors of dendritic cells (DCs), macrophages and granulocytes. MDSCs proliferate in the blood, spleen, and tumor tissues in tumorbearing mice and in the peripheral blood and tumor tissues in patients with cancer. MDSCs prevent tumors from attacks by body immunosurveillance and promote tumors progression through inhibiting both innate and adaptive antitumor immunity by a variety of pathways; they are recruited to the peripheral tissues from bone marrow and exert their inhibitory effects on antitumor immunity after activation in peripheral tissues. Chronic inflammationrelated cytokines produced by tumors play crucial roles in the recruitment and activation of MDSCs. Progress has been made in antitumor therapies targeting MDSCs. But it has only been 10 years since the discovery of MDSCs, and many questions remain to be answered through experimental and clinical investigations. This review focuses on progress in MDSCs and its subsets, the recruitment and activation of MDSCs, the mechanisms of MDSCsmediated immunosurveillance and antitumor treatment targeting MDSCs.
Myeloidderived suppressor cells (MDSCs) are heterogeneous cells derived from myeloid progenitor cells and immature myeloid cells (IMCs) in bone marrow; they are the progenitors of dendritic cells (DCs), macrophages and granulocytes. MDSCs proliferate in the blood, spleen, and tumor tissues in tumorbearing mice and in the peripheral blood and tumor tissues in patients with cancer. MDSCs prevent tumors from attacks by body immunosurveillance and promote tumors progression through inhibiting both innate and adaptive antitumor immunity by a variety of pathways; they are recruited to the peripheral tissues from bone marrow and exert their inhibitory effects on antitumor immunity after activation in peripheral tissues. Chronic inflammationrelated cytokines produced by tumors play crucial roles in the recruitment and activation of MDSCs. Progress has been made in antitumor therapies targeting MDSCs. But it has only been 10 years since the discovery of MDSCs, and many questions remain to be answered through experimental and clinical investigations. This review focuses on progress in MDSCs and its subsets, the recruitment and activation of MDSCs, the mechanisms of MDSCsmediated immunosurveillance and antitumor treatment targeting MDSCs.
2009, 16(4):325-330.
DOI: 10.3872/j.issn.1007-385X.2009.4.002
Abstract:
Objective:To investigate the in vitro effect of icariin (ICA) on the differentiation, maturation and immune activity of DCs derived from cord blood mononuclear cells.Methods: The mononuclear cells were isolated by density gradient centrifugation from cord blood under axenic condition, and were induced by GMCSF and IL4 to differentiate into DCs. The DCs were divided into 3 groups on the fifth day: an ICA group, a TNFα group and a control group. The morphology of DCs was observed under inverted microscope and transmission election microscope. Expression of CD83, CD80 and CD86 on DCs was examined by flow cytometry. IL12 and IFNγ levels in the culture supernatant of DCs were detected by ELISA. The proliferation of T cells stimulated by DCs was determined by MTT assay.Results: Cord blood mononuclear cells derivedDCs had a typical morphological characteristic of mature DCs after stimulation with ICA. Expression of CD1a, CD83, CD80 and CD86 on DCs in ICA group was upregulated compared with that in control group (P<0.05); moreover, CD86 expression on DCs in ICA group was significantly higher than that in TNFα group (P<0.05). IL12 and IFNγ production in ICA group and proliferation of T cells induced by DCs in the ICA group were markedly higher than those in the control group (P<0.05), and were similar to those in the TNFα group.Conclusion: ICA can induce differentiation and maturation of DCs derived from cord blood mononuclear cells, and enhances the immune activity of DCs.
Objective:To investigate the in vitro effect of icariin (ICA) on the differentiation, maturation and immune activity of DCs derived from cord blood mononuclear cells.Methods: The mononuclear cells were isolated by density gradient centrifugation from cord blood under axenic condition, and were induced by GMCSF and IL4 to differentiate into DCs. The DCs were divided into 3 groups on the fifth day: an ICA group, a TNFα group and a control group. The morphology of DCs was observed under inverted microscope and transmission election microscope. Expression of CD83, CD80 and CD86 on DCs was examined by flow cytometry. IL12 and IFNγ levels in the culture supernatant of DCs were detected by ELISA. The proliferation of T cells stimulated by DCs was determined by MTT assay.Results: Cord blood mononuclear cells derivedDCs had a typical morphological characteristic of mature DCs after stimulation with ICA. Expression of CD1a, CD83, CD80 and CD86 on DCs in ICA group was upregulated compared with that in control group (P<0.05); moreover, CD86 expression on DCs in ICA group was significantly higher than that in TNFα group (P<0.05). IL12 and IFNγ production in ICA group and proliferation of T cells induced by DCs in the ICA group were markedly higher than those in the control group (P<0.05), and were similar to those in the TNFα group.Conclusion: ICA can induce differentiation and maturation of DCs derived from cord blood mononuclear cells, and enhances the immune activity of DCs.
2009, 16(4):331-335.
DOI: 10.3872/j.issn.1007-385X.2009.4.003
Abstract:
Objective:To investigate the effect of APE1targeting siRNA (APE1 siRNA) on implanted osteosarcoma and its synergetic role with bevacizumab (vascular endothelial growth factor antibody, Avastin). Methods: Human osteosarcoma 9901 cellbearing nude mouse model was established. Sixteen tumorbearing mice were randomly divided into 4 groups: EGFP control group, APE1 siRNA group, Avastin group and combined treatment group (Avastin+APE1 siRNA). The growth of implanted tumors was measured during treatment, and tumor inhibitotry rate was calculated. Meanwhile, microvessel density (MVD) and Ki67 expression in tumor tissues were examined by immunohistochemistry. Apoptosis of tumor cells was examined by TUNEL. Hypoxia status of tumor tissues was determined by laser cofocal scanning microscopy. Expression of VEGF protein in tumor tissues was detected by Western blotting. Results: The tumor inhibitory rate of Avastin+APE1 siRNA group was significantly increased compared with those of APE1 siRNA group and Avastin group (P<0.01). The MVD and Ki67 expressions in therapy groups were significantly lower than those in control group; moreover, the MVD and Ki67 expressions in Avastin+APE1 siRNA group were remarkedly lower than those in APE1 siRNA group or Avastin group (P<0.01). The apoptosis index (AI) in therapy groups was significantly higher than that in control group, with AI in Avastin+APE1 siRNA group being markedly higher than those in the other two groups (P<0.01). Hypoxia status in tumor tissues was enhanced and VEGF expression was inhibited in APE1 siRNA group or Avastin group; furthermore, hypoxia status and VEGF expression in Avastin+APE1 siRNA group were greatly changed.Conclusion: APE1targeting siRNA can inhibit the angiogenesis and growth of implanted osterosarcoma, and induce apoptosis of tumor cells, which shows a synergistic role with Avastin in the treatment of osteosarcoma.
Objective:To investigate the effect of APE1targeting siRNA (APE1 siRNA) on implanted osteosarcoma and its synergetic role with bevacizumab (vascular endothelial growth factor antibody, Avastin). Methods: Human osteosarcoma 9901 cellbearing nude mouse model was established. Sixteen tumorbearing mice were randomly divided into 4 groups: EGFP control group, APE1 siRNA group, Avastin group and combined treatment group (Avastin+APE1 siRNA). The growth of implanted tumors was measured during treatment, and tumor inhibitotry rate was calculated. Meanwhile, microvessel density (MVD) and Ki67 expression in tumor tissues were examined by immunohistochemistry. Apoptosis of tumor cells was examined by TUNEL. Hypoxia status of tumor tissues was determined by laser cofocal scanning microscopy. Expression of VEGF protein in tumor tissues was detected by Western blotting. Results: The tumor inhibitory rate of Avastin+APE1 siRNA group was significantly increased compared with those of APE1 siRNA group and Avastin group (P<0.01). The MVD and Ki67 expressions in therapy groups were significantly lower than those in control group; moreover, the MVD and Ki67 expressions in Avastin+APE1 siRNA group were remarkedly lower than those in APE1 siRNA group or Avastin group (P<0.01). The apoptosis index (AI) in therapy groups was significantly higher than that in control group, with AI in Avastin+APE1 siRNA group being markedly higher than those in the other two groups (P<0.01). Hypoxia status in tumor tissues was enhanced and VEGF expression was inhibited in APE1 siRNA group or Avastin group; furthermore, hypoxia status and VEGF expression in Avastin+APE1 siRNA group were greatly changed.Conclusion: APE1targeting siRNA can inhibit the angiogenesis and growth of implanted osterosarcoma, and induce apoptosis of tumor cells, which shows a synergistic role with Avastin in the treatment of osteosarcoma.
2009, 16(4):336-341.
DOI: 10.3872/j.issn.1007-385X.2009.4.004
Abstract:
Objective:To express Kininogen D560148TRAIL114281 fusion protein using prokaryotic system and observe its biological functions.Methods: The Kininogen D560148 gene and TNFrelated apoptosisinducing ligand (TRAIL114281 ) gene were amplified by PCR and were cloned into pMAL expression vector to construct recombinant pMALKininogen D560148 (pMALKD5), pMALTRAIL114281 (pMALTRAIL) and pMALKininogen D560148TRAIL114281 (pMALKT) plasmids, respectively. The plasmids were transformed into E. coli BL21 and were efficiently expressed after IPTG induction. The purified MBPKD5, MBPTRAIL and MBPKT proteins were obtained by amylose resin affinity purification column. The proliferation of cells was measured by MTT; tube formation of endothelial cell was detected by tube formation assay; and the apoptosis of cells were observed by electron microscopic and FCM.Results: Prokaryotic expression vectors pMALKD5, pMALTRAIL and pMALKT and their purified fusion proteins MBPKD5, MBPTRAIL and MBPKT were successfully obtained. MBPKT significantly inhibited the proliferation of ECV304 endothelial cells, SW1990 pancreatic cancer cells and the tube formation of ECV304 cells compared with those of MBPKD5 and MBPTRAIL. Meanwhile, MBPKT dosedependently induced the apoptosis of SW1990 cells.Conclusion: Kininogen D560148TRAIL114281 fusion protein can inhibit the proliferation of tumor cells and angiogenesis of endothelial cells, which lays a foundation for further research on tumortargeting drugs.
Objective:To express Kininogen D560148TRAIL114281 fusion protein using prokaryotic system and observe its biological functions.Methods: The Kininogen D560148 gene and TNFrelated apoptosisinducing ligand (TRAIL114281 ) gene were amplified by PCR and were cloned into pMAL expression vector to construct recombinant pMALKininogen D560148 (pMALKD5), pMALTRAIL114281 (pMALTRAIL) and pMALKininogen D560148TRAIL114281 (pMALKT) plasmids, respectively. The plasmids were transformed into E. coli BL21 and were efficiently expressed after IPTG induction. The purified MBPKD5, MBPTRAIL and MBPKT proteins were obtained by amylose resin affinity purification column. The proliferation of cells was measured by MTT; tube formation of endothelial cell was detected by tube formation assay; and the apoptosis of cells were observed by electron microscopic and FCM.Results: Prokaryotic expression vectors pMALKD5, pMALTRAIL and pMALKT and their purified fusion proteins MBPKD5, MBPTRAIL and MBPKT were successfully obtained. MBPKT significantly inhibited the proliferation of ECV304 endothelial cells, SW1990 pancreatic cancer cells and the tube formation of ECV304 cells compared with those of MBPKD5 and MBPTRAIL. Meanwhile, MBPKT dosedependently induced the apoptosis of SW1990 cells.Conclusion: Kininogen D560148TRAIL114281 fusion protein can inhibit the proliferation of tumor cells and angiogenesis of endothelial cells, which lays a foundation for further research on tumortargeting drugs.
2009, 16(4):342-346.
DOI: 10.3872/j.issn.1007-385X.2009.4.005
Abstract:
Objective:To investigate the effects of Ki67targeting peptide nucleic acids (PNAs) linked with dihydrotestosterone (DHT) on the Ki67 expression, proliferation and apoptosis of hormoneindependent prostate cancer cell line PC3. Methods: Ki67targeting PNAs were artificially synthesized and were covalently linked to DHT to yield DHTPNAs. DHTPNAs were then transfected into PC3 cells. Ki67 expression in PC3 cells was examined by RTPCR, immunohistochemistry and Western blotting assay. The proliferation of PC3 cells was examined by CCK8 assay, and the apoptosis of PC3 cells was detected by TUNEL assay. Groups treated with PNAs or DHT served as controls. Results: Both PNAs and DHTPNAs inhibited the expression of Ki67 in PC3 cells, increased the apoptotsis of PC3 cells, and inhibited the proliferation of PC3 cells in a dosedependent manner. The therapeutic effect of DHTPNAs at the same concentration was better than that of PNAs, with 3 μmol/L DHTPNAs (onethird of PNAs dose) reaching the same therapeutic effect of PNAs.Conclusion: DHTPNAs can promote the effects of PNAs in inhibiting Ki67 expression, inducing apoptosis and inhibiting proliferation of PC3 cells.
Objective:To investigate the effects of Ki67targeting peptide nucleic acids (PNAs) linked with dihydrotestosterone (DHT) on the Ki67 expression, proliferation and apoptosis of hormoneindependent prostate cancer cell line PC3. Methods: Ki67targeting PNAs were artificially synthesized and were covalently linked to DHT to yield DHTPNAs. DHTPNAs were then transfected into PC3 cells. Ki67 expression in PC3 cells was examined by RTPCR, immunohistochemistry and Western blotting assay. The proliferation of PC3 cells was examined by CCK8 assay, and the apoptosis of PC3 cells was detected by TUNEL assay. Groups treated with PNAs or DHT served as controls. Results: Both PNAs and DHTPNAs inhibited the expression of Ki67 in PC3 cells, increased the apoptotsis of PC3 cells, and inhibited the proliferation of PC3 cells in a dosedependent manner. The therapeutic effect of DHTPNAs at the same concentration was better than that of PNAs, with 3 μmol/L DHTPNAs (onethird of PNAs dose) reaching the same therapeutic effect of PNAs.Conclusion: DHTPNAs can promote the effects of PNAs in inhibiting Ki67 expression, inducing apoptosis and inhibiting proliferation of PC3 cells.
RONG Tingting
,
CHEN Huijuan
,
ZHANG Huizhen
,
LIU Zhengchun
,
ZHANG Yong
,
WANG Shujun
,
WANG Ying
,
GE Hailiang
2009, 16(4):347-352.
DOI: 10.3872/j.issn.1007-385X.2009.4.006
Abstract:
Objective:To prepare and characterize monoclonal antibody against tumorassociated antigen OVA66, so as to provide effective method for further studying the biological function of OVA66. Methods: Recombinant pET32bOVA66 plasmid was constructed and transfected into E.coli BL21 (DE3). HisOVA66 fusion protein was induced by IPTG and purified through NiTED affinity chromatography. Hybridoma cells stably secreting antiOVA66 monoclonal antibodies were prepared by hybridoma technology. Biological and immunologic properties of monoclonal antibodies were determined by ELISA and Western blotting. The prepared antibody was applied in immunofluorescence, flow cytometry and immunohistochemistry procedures. Results: Recombinant pET32bOVA66 plasmid was successfully constructed, and OVA66 fusion protein was induced and further purified. Two hybridoma cell strains secreting OVA66 monoclonal antibodies were established by hybridoma technology and named as 5F4 and 4G9, respectively. These monoclonal antibodies were characterized as IgG1 subclass and kappa subtype with affinity constant ka being 2.96×1010 and 0.4×1010L/mol, respectively. Preliminary data showed that these antibodies acted against different antigenic epitopes. These antibodies could be used for the detection of OVA66 expression in immunofluorescence and flow cytometry assay, whereas 5F4 could also be used in immunohistochemistry assay. Conclusion: Two hybridoma cell strains secreting monoclonal antibodies against OVA66 have been successfully established, which facilitates further studies on biological functions of OVA66 and its potential in clinical application.
Objective:To prepare and characterize monoclonal antibody against tumorassociated antigen OVA66, so as to provide effective method for further studying the biological function of OVA66. Methods: Recombinant pET32bOVA66 plasmid was constructed and transfected into E.coli BL21 (DE3). HisOVA66 fusion protein was induced by IPTG and purified through NiTED affinity chromatography. Hybridoma cells stably secreting antiOVA66 monoclonal antibodies were prepared by hybridoma technology. Biological and immunologic properties of monoclonal antibodies were determined by ELISA and Western blotting. The prepared antibody was applied in immunofluorescence, flow cytometry and immunohistochemistry procedures. Results: Recombinant pET32bOVA66 plasmid was successfully constructed, and OVA66 fusion protein was induced and further purified. Two hybridoma cell strains secreting OVA66 monoclonal antibodies were established by hybridoma technology and named as 5F4 and 4G9, respectively. These monoclonal antibodies were characterized as IgG1 subclass and kappa subtype with affinity constant ka being 2.96×1010 and 0.4×1010L/mol, respectively. Preliminary data showed that these antibodies acted against different antigenic epitopes. These antibodies could be used for the detection of OVA66 expression in immunofluorescence and flow cytometry assay, whereas 5F4 could also be used in immunohistochemistry assay. Conclusion: Two hybridoma cell strains secreting monoclonal antibodies against OVA66 have been successfully established, which facilitates further studies on biological functions of OVA66 and its potential in clinical application.
2009, 16(4):353-357.
DOI: 10.3872/j.issn.1007-385X.2009.4.007
Abstract:
Objective:To investigate the effect of P53 upregulate modulator of apoptosis (PUMA) on the apoptosis of pancreatic carcinoma BxPC3 cells and the possible mechanism. Methods: BxPC3 cells were infected with recombinant adenovirus containing PUMA gene (AdPUMA) at 100 MOI for 096 h. Apoptosis of BxPC3 cells was examined by FCM. Expressions of PUMA, Bcl2, Bax, Cytochrome C and Caspase3 proteins in BxPC3 cells were detected by Western blotting. Bax expression in the cytoplasm and mitochondrion and Bax oligomer expression expression in BxPC3 cells were determined by Western blotting. Results:Apoptosis rates of BxPC3 cells were significantly increased with the time of AdPUMA infection, and peaked after 48 h. AdPUMA infection increased the expressions of PUMA, Cytochrome C and Caspase3 proteins in BxPC3 cells, and decreased the expression of Bcl2 protein. Apoptosis rate of BxPC3 cells after AdPUMA infection was correlated with PUMA expression. AdPUMA did not affect the expression of total Bax protein in BxPC3 cells, but Bax expression in cytoplasm was dramatically decreased after infection, and Bax expression in mitochondrion was markedly increased. Furthermore, AdPUMA infection induced Bax oligomerization in BxPC3 cells.Conclusion: PUMA can promote apoptosis of pancreatic carcinoma cells through mitochondrion pathway.
Objective:To investigate the effect of P53 upregulate modulator of apoptosis (PUMA) on the apoptosis of pancreatic carcinoma BxPC3 cells and the possible mechanism. Methods: BxPC3 cells were infected with recombinant adenovirus containing PUMA gene (AdPUMA) at 100 MOI for 096 h. Apoptosis of BxPC3 cells was examined by FCM. Expressions of PUMA, Bcl2, Bax, Cytochrome C and Caspase3 proteins in BxPC3 cells were detected by Western blotting. Bax expression in the cytoplasm and mitochondrion and Bax oligomer expression expression in BxPC3 cells were determined by Western blotting. Results:Apoptosis rates of BxPC3 cells were significantly increased with the time of AdPUMA infection, and peaked after 48 h. AdPUMA infection increased the expressions of PUMA, Cytochrome C and Caspase3 proteins in BxPC3 cells, and decreased the expression of Bcl2 protein. Apoptosis rate of BxPC3 cells after AdPUMA infection was correlated with PUMA expression. AdPUMA did not affect the expression of total Bax protein in BxPC3 cells, but Bax expression in cytoplasm was dramatically decreased after infection, and Bax expression in mitochondrion was markedly increased. Furthermore, AdPUMA infection induced Bax oligomerization in BxPC3 cells.Conclusion: PUMA can promote apoptosis of pancreatic carcinoma cells through mitochondrion pathway.
2009, 16(4):358-363.
DOI: 10.3872/j.issn.1007-385X.2009.4.008
Abstract:
Objective:To study the effects of transcription factor activator protein2α (AP2α）on invasive growth and estrogen receptorβ (ERβ) expression in human colon cancer SW620 cells, and to probe into the involved molecular mechanism.Methods: Plasmid pcDNA3.1(＋) AP2α and pcDNA3.1(＋) were transfected into SW620 cells by liposomemediated transfection. The adhesion, invasion and migration abilities of SW620 cells were measured by metrical gel adhesion assay and modified Boyden chamber (Transwell assay). The gene and protein expression levels of AP2α and ERβ in SW620 cells were examined by Realtime PCR, Western blotting and immunofluorescence cytochemistry. The interaction between AP2α DNA and ERβ in SW620 cells was measured by electrophoretic mobility shift assay (EMSA) afterAP2αgene transfection.Results: Overexpression of AP2α markedly reduced the adhesion, invasion and migration abilities of SW620 cells (all P<0.05); meanwhile, the mRNA and protein levels of ERβ in SW620 cells were also significantly enhanced (P<0.05). EMSA results showed that AP2α specifically bound to promoter region of ERβ gene in SW620 cells after transfection of pcDNA3.1(+)AP2αplasmid. Conclusion: Overexpression of AP2α can inhibit the adhesion, invasion and migration abilities of SW620 cells, which is probably related to ERβ expression in SW620 cells directly induced by interaction between AP2α and promoter region of ERβ.
Objective:To study the effects of transcription factor activator protein2α (AP2α）on invasive growth and estrogen receptorβ (ERβ) expression in human colon cancer SW620 cells, and to probe into the involved molecular mechanism.Methods: Plasmid pcDNA3.1(＋) AP2α and pcDNA3.1(＋) were transfected into SW620 cells by liposomemediated transfection. The adhesion, invasion and migration abilities of SW620 cells were measured by metrical gel adhesion assay and modified Boyden chamber (Transwell assay). The gene and protein expression levels of AP2α and ERβ in SW620 cells were examined by Realtime PCR, Western blotting and immunofluorescence cytochemistry. The interaction between AP2α DNA and ERβ in SW620 cells was measured by electrophoretic mobility shift assay (EMSA) afterAP2αgene transfection.Results: Overexpression of AP2α markedly reduced the adhesion, invasion and migration abilities of SW620 cells (all P<0.05); meanwhile, the mRNA and protein levels of ERβ in SW620 cells were also significantly enhanced (P<0.05). EMSA results showed that AP2α specifically bound to promoter region of ERβ gene in SW620 cells after transfection of pcDNA3.1(+)AP2αplasmid. Conclusion: Overexpression of AP2α can inhibit the adhesion, invasion and migration abilities of SW620 cells, which is probably related to ERβ expression in SW620 cells directly induced by interaction between AP2α and promoter region of ERβ.
2009, 16(4):364-368.
DOI: 10.3872/j.issn.1007-385X.2009.4.009
Abstract:
Objective:To establish the quality control methods and standards for recombinant antitumorantivirusprotein novaferon. Methods: Antitumor activity of novaferon was evaluated by determining its inhibitory effect on the proliferation of Daudi cells using WST1 staining. The antivirus activity of novaferon was examined by cytopathic inhibition assay on WISH cells. The peptide map of novaferon was obtained by trypsin digestion and HPLC assay. Other routine tests were all performed according to the pharmacopoeia of the People′s Republic of China.Results: The antitumor and antivirus activities of three batches of novaferon bulk and final products were ≥2.0×106 U/mg and ≥1.0×109 IU/mg, respectively. HPLC peptide maps of three batches of novaferon paralleled with that of standard product. Quantity, purity, molecular mass, isoelectic point and Nterminal amino acid sequence of three batches of novaferon bulk products and bacterial endotoxin content, benzyl alcohol content, acetonitrile of three final products were consistent with the quality standards. Results of other routine tests all complied with the standard requirements. The quality control methods and standards for recombinant antitumorantivirusprotein novaferon were established accordingly.Conclusion: The established quality control methods and standards for novaferon are safe and effective, which can be used for routine quality control of novaferon.
Objective:To establish the quality control methods and standards for recombinant antitumorantivirusprotein novaferon. Methods: Antitumor activity of novaferon was evaluated by determining its inhibitory effect on the proliferation of Daudi cells using WST1 staining. The antivirus activity of novaferon was examined by cytopathic inhibition assay on WISH cells. The peptide map of novaferon was obtained by trypsin digestion and HPLC assay. Other routine tests were all performed according to the pharmacopoeia of the People′s Republic of China.Results: The antitumor and antivirus activities of three batches of novaferon bulk and final products were ≥2.0×106 U/mg and ≥1.0×109 IU/mg, respectively. HPLC peptide maps of three batches of novaferon paralleled with that of standard product. Quantity, purity, molecular mass, isoelectic point and Nterminal amino acid sequence of three batches of novaferon bulk products and bacterial endotoxin content, benzyl alcohol content, acetonitrile of three final products were consistent with the quality standards. Results of other routine tests all complied with the standard requirements. The quality control methods and standards for recombinant antitumorantivirusprotein novaferon were established accordingly.Conclusion: The established quality control methods and standards for novaferon are safe and effective, which can be used for routine quality control of novaferon.
2009, 16(4):369-373.
DOI: 10.3872/j.issn.1007-385X.2009.4.010
Abstract:
Objective:To observe the inhibitory effect of gefitinib combined with DNA vaccine targeting EGFR against implanted Lewis tumors in mice.Methods: Chicken EGFR L2 domain and rabbit IgG Fc domain fusion pVAX1/cEGFRrFc DNA vaccine was injected into mice and the titer of antiEGFR in serum was determined by ELISA. The growth of Lewis cells was measured by MTT. Lewis lung cancer mouse models were established and were randomly divided into vaccine, gefitinib, gefitinib+vaccine, and control groups. The tumor volume and weight and survival of mice were examined in different groups.Results:The titer of antiEGFR in mice vaccinated with pVAX1/cEGFRrFc plasmid was 1∶1 000. The proliferation of Lewis cells in antiEGFR combined with gefitinib was significantly inhibited compared with those in antiEGFR and gefitinib groups (P<0.05). In vivo results showed that the growth of implanted Lewis tumors in vaccine+gefitinib group was greatly inhibited, and the survival rate of Lewisbearing mice was significalty increased (P<001). Conclusion：DNA vaccine targeting EGFR has a synergistic effect with gefitinib in antitumor activity, and this DNA vaccine can increase the efficiency of gefitinib.
Objective:To observe the inhibitory effect of gefitinib combined with DNA vaccine targeting EGFR against implanted Lewis tumors in mice.Methods: Chicken EGFR L2 domain and rabbit IgG Fc domain fusion pVAX1/cEGFRrFc DNA vaccine was injected into mice and the titer of antiEGFR in serum was determined by ELISA. The growth of Lewis cells was measured by MTT. Lewis lung cancer mouse models were established and were randomly divided into vaccine, gefitinib, gefitinib+vaccine, and control groups. The tumor volume and weight and survival of mice were examined in different groups.Results:The titer of antiEGFR in mice vaccinated with pVAX1/cEGFRrFc plasmid was 1∶1 000. The proliferation of Lewis cells in antiEGFR combined with gefitinib was significantly inhibited compared with those in antiEGFR and gefitinib groups (P<0.05). In vivo results showed that the growth of implanted Lewis tumors in vaccine+gefitinib group was greatly inhibited, and the survival rate of Lewisbearing mice was significalty increased (P<001). Conclusion：DNA vaccine targeting EGFR has a synergistic effect with gefitinib in antitumor activity, and this DNA vaccine can increase the efficiency of gefitinib.
2009, 16(4):374-378.
DOI: 10.3872/j.issn.1007-385X.2009.4.011
Abstract:
Objective: To observe the effect of RNAi targeting survivin gene (RNAisurvivin) on the growth and apoptosis of implanted human cervical carcinoma in mice. Methods: Nude mice were randomly divided into 4 groups, and the human cervical carcinoma HeLa cells transfected with pSilencer2.1s2, pSilencer2.1NC, pSilencer2.1U6 neo or not transfected (the cells being HeLas2, HeLaNC, HeLaU6 neo and Hela, respectively) were subcutaneously inoculated into flank tissues of mice to establish cervical carcinoma mouse models. The influence of survivinRNAi on the implanted tumor growth was observed. Expression of survivin protein and microvessel density (MVD) in implanted cervical carcinoma tissues were examined by immunohistochemistry. Apoptosis of implanted cervical carcinoma cells was observed by HE staining and TUNEL. Results: Implanted human cervical carcinoma mouse model was successively established. The tumor weight in HeLas2 group was markedly lower than those in HeLaNC, HeLaU6 neo and Hela groups（P<0.05）. The tumor inhibitory rate in HeLas2 group was 67.9%. Survivin expression and MVD value in tumor tissues of HeLas2 group were significantly decreased as determined by immunohistochemistry（all P<0.05）. Apoptosis of implanted cervical carcinoma cells in HeLas2 group was significantly increased compared with those in other 3 groups（all P<0.05）, with AI being (22.73±1.37)%. Conclusion:RNAisurvivin inhibits the growth and promote apoptosis of implanted human cervical carcinoma tumors by downregulating survivin protein expression and MVD in tumor tissues.
Objective: To observe the effect of RNAi targeting survivin gene (RNAisurvivin) on the growth and apoptosis of implanted human cervical carcinoma in mice. Methods: Nude mice were randomly divided into 4 groups, and the human cervical carcinoma HeLa cells transfected with pSilencer2.1s2, pSilencer2.1NC, pSilencer2.1U6 neo or not transfected (the cells being HeLas2, HeLaNC, HeLaU6 neo and Hela, respectively) were subcutaneously inoculated into flank tissues of mice to establish cervical carcinoma mouse models. The influence of survivinRNAi on the implanted tumor growth was observed. Expression of survivin protein and microvessel density (MVD) in implanted cervical carcinoma tissues were examined by immunohistochemistry. Apoptosis of implanted cervical carcinoma cells was observed by HE staining and TUNEL. Results: Implanted human cervical carcinoma mouse model was successively established. The tumor weight in HeLas2 group was markedly lower than those in HeLaNC, HeLaU6 neo and Hela groups（P<0.05）. The tumor inhibitory rate in HeLas2 group was 67.9%. Survivin expression and MVD value in tumor tissues of HeLas2 group were significantly decreased as determined by immunohistochemistry（all P<0.05）. Apoptosis of implanted cervical carcinoma cells in HeLas2 group was significantly increased compared with those in other 3 groups（all P<0.05）, with AI being (22.73±1.37)%. Conclusion:RNAisurvivin inhibits the growth and promote apoptosis of implanted human cervical carcinoma tumors by downregulating survivin protein expression and MVD in tumor tissues.
2009, 16(4):379-382.
DOI: 10.3872/j.issn.1007-385X.2009.4.012
Abstract:
Objective:To study the effects of IL2 and IL15 on the expression of NKG2D and the cytotoxicity of editedNK cells against human nasopharyngeal carcinoma cell line CNE2. Methods: NK cells were purified by antiCD56 MACS and were divided into four groups: noneditedNK cells group (NK cells treated with 100 U/ml IL2), editedNK cells group (NK cells cocultured with CNE2 cells at a ratio of 10∶1 and then treated with 100 U/ml IL2), editedNK cells retreated with 1 000 U/ml IL2 group, and editedNK cells retreated with 10 ng/ml IL15 group. Expression of NKG2D in each group was determined by FACS 24 h later. Cytotoxicity of NK cells against CNE2 cells (NK∶CNE2 being 20∶1) was measured by LDH releasing assay.Results: The expression of NKG2D in noneditedNK cells, editedNK cells, editedNK cells retreated with IL2, and editedNK cells retreated with IL15 were (97.63±0.83)%, (53.50±1.25)%, (94.47±1.00)%, and (98.07±0.21)%, respectively. The expression of NKG2D on editedNK cells retreated with IL2 or IL15 was significantly increased than that on editedNK cells (P<0.01). The cytotoxicity of noneditedNK cells, editedNK cells, editedNK cells retreated with IL2, and editedNK cells retreated with IL15 against CNE2 cells were (35.90±3.27)%, (4.70±2.30)%, (31.70 ±3.56)% and (40.18±2.94)%, respectively. The cytotoxicity of editedNK cells was significantly enhanced after retreated with IL2 or IL15 (P<0.01), with those retreated with IL15 being stronger than those retreated with IL2.Conclusion: High dose IL2 and IL15 can upregulate the expression of NKG2D on editedNK cells and restore their cytotoxicity against CNE2 cells, with the efficacy of IL15 stronger than that of IL2.
Objective:To study the effects of IL2 and IL15 on the expression of NKG2D and the cytotoxicity of editedNK cells against human nasopharyngeal carcinoma cell line CNE2. Methods: NK cells were purified by antiCD56 MACS and were divided into four groups: noneditedNK cells group (NK cells treated with 100 U/ml IL2), editedNK cells group (NK cells cocultured with CNE2 cells at a ratio of 10∶1 and then treated with 100 U/ml IL2), editedNK cells retreated with 1 000 U/ml IL2 group, and editedNK cells retreated with 10 ng/ml IL15 group. Expression of NKG2D in each group was determined by FACS 24 h later. Cytotoxicity of NK cells against CNE2 cells (NK∶CNE2 being 20∶1) was measured by LDH releasing assay.Results: The expression of NKG2D in noneditedNK cells, editedNK cells, editedNK cells retreated with IL2, and editedNK cells retreated with IL15 were (97.63±0.83)%, (53.50±1.25)%, (94.47±1.00)%, and (98.07±0.21)%, respectively. The expression of NKG2D on editedNK cells retreated with IL2 or IL15 was significantly increased than that on editedNK cells (P<0.01). The cytotoxicity of noneditedNK cells, editedNK cells, editedNK cells retreated with IL2, and editedNK cells retreated with IL15 against CNE2 cells were (35.90±3.27)%, (4.70±2.30)%, (31.70 ±3.56)% and (40.18±2.94)%, respectively. The cytotoxicity of editedNK cells was significantly enhanced after retreated with IL2 or IL15 (P<0.01), with those retreated with IL15 being stronger than those retreated with IL2.Conclusion: High dose IL2 and IL15 can upregulate the expression of NKG2D on editedNK cells and restore their cytotoxicity against CNE2 cells, with the efficacy of IL15 stronger than that of IL2.
2009, 16(4):383-386.
DOI: 10.3872/j.issn.1007-385X.2009.4.013
Abstract:
Objective:To study the effect of siRNA targeting osteopontin（OPNRNAi）on the proliferation and invasiveness of U87 glioma cells and the possible mechanism. Methods: OPNRNAi was synthesized according to the gene sequence of OPN protein and was transfected into U87 cells. The proliferation of U87 cells was examined by MTT; matrix metalloprotease (MMP) 2 and MMP9 expression were detected by Western blotting assay; transwell assay and gelatinzymogram were used to detect the invasion ability of U87 cells and gelatinase acitivity of MMP2 and MMP9, respectively.Results:Synthesized OPNRNAi effectively inhibited the expression of OPN in U87 cells (P<0.05). OPNRNAi also significantly decreased the expression of MMP2 and MMP9 in U87 cells (P<0.05) and the gelation activity of MMP2 and MMP9 (P<0.01), and inhibited the proliferation and invasivenss of U87 cells（all P<0.05）. Conclusion: Knockdown of OPN with OPNRNAi can inhibit the proliferation and invasiveness of U87 cells, which is probably related to the decreased expression of MMP2 and MMP9 genes and their gelatinase acitivities.
Objective:To study the effect of siRNA targeting osteopontin（OPNRNAi）on the proliferation and invasiveness of U87 glioma cells and the possible mechanism. Methods: OPNRNAi was synthesized according to the gene sequence of OPN protein and was transfected into U87 cells. The proliferation of U87 cells was examined by MTT; matrix metalloprotease (MMP) 2 and MMP9 expression were detected by Western blotting assay; transwell assay and gelatinzymogram were used to detect the invasion ability of U87 cells and gelatinase acitivity of MMP2 and MMP9, respectively.Results:Synthesized OPNRNAi effectively inhibited the expression of OPN in U87 cells (P<0.05). OPNRNAi also significantly decreased the expression of MMP2 and MMP9 in U87 cells (P<0.05) and the gelation activity of MMP2 and MMP9 (P<0.01), and inhibited the proliferation and invasivenss of U87 cells（all P<0.05）. Conclusion: Knockdown of OPN with OPNRNAi can inhibit the proliferation and invasiveness of U87 cells, which is probably related to the decreased expression of MMP2 and MMP9 genes and their gelatinase acitivities.
2009, 16(4):387-390.
DOI: 10.3872/j.issn.1007-385X.2009.4.014
Abstract:
Objective:To investigate the effectiveness of tissue specific cytisine deaminase/5fluorocytosine (CD/5FC) thermochemotherapy system in treatment of liver metastasis of colon cancer in nude mice. Methods: CEA promoterregulated recombinant retroviral vector G1CEACDNa was packaged, propagated and purified, and the viral supernatant was harvested. Human colon cancer LoVo cells were injected into the portal veins of 45 nude mice. Two days after the establishment of liver metastasis model, the viral supernatant was intraperitoneally injected into mice（0.2 ml/d, 5 d）. The 45 mice were then randomly divided into 3 groups, namely, the control group (injected with sodium), chemotherapy group (prodrug/5FC) and thermochemotherapy group (43 ℃ prodrug/5FC). After treated for 21 d, the mice were sacrificed and liver metastasis rate and liver metastasis nodule numbers were observed. Expression of CD gene in liver metastasis tissues was determined by RTPCR. Pathological changes of liver metastasis tissues were examined by light microscope and electron microscope.Results: The virus titer of G1CEACDNa was 5.6×106 CFU/L. CD gene was effectively expressed in the liver metastasis tissues. Liver metastasis rates and number of liver metastasis nodules were significantly lower in the thermochemotherapy group than in the chemotherapy group (13.3% vs 400%, \[0.20±0.56\] vs \[0.80±1.01\]; all P<0.05). The tumor cells grew well in the control group, and were greatly inhibited in the thermochemotherapy group compared that in the other two groups. The tumor cells showed different degrees of apoptosis in the thermochemotherapy and chemotherapy groups under electron microscope.Conclusion: The tissue specific CD/5FC thermochemotherapy system can inhibit the growth of liver metastasis of colon cancer in nude mice.
Objective:To investigate the effectiveness of tissue specific cytisine deaminase/5fluorocytosine (CD/5FC) thermochemotherapy system in treatment of liver metastasis of colon cancer in nude mice. Methods: CEA promoterregulated recombinant retroviral vector G1CEACDNa was packaged, propagated and purified, and the viral supernatant was harvested. Human colon cancer LoVo cells were injected into the portal veins of 45 nude mice. Two days after the establishment of liver metastasis model, the viral supernatant was intraperitoneally injected into mice（0.2 ml/d, 5 d）. The 45 mice were then randomly divided into 3 groups, namely, the control group (injected with sodium), chemotherapy group (prodrug/5FC) and thermochemotherapy group (43 ℃ prodrug/5FC). After treated for 21 d, the mice were sacrificed and liver metastasis rate and liver metastasis nodule numbers were observed. Expression of CD gene in liver metastasis tissues was determined by RTPCR. Pathological changes of liver metastasis tissues were examined by light microscope and electron microscope.Results: The virus titer of G1CEACDNa was 5.6×106 CFU/L. CD gene was effectively expressed in the liver metastasis tissues. Liver metastasis rates and number of liver metastasis nodules were significantly lower in the thermochemotherapy group than in the chemotherapy group (13.3% vs 400%, \[0.20±0.56\] vs \[0.80±1.01\]; all P<0.05). The tumor cells grew well in the control group, and were greatly inhibited in the thermochemotherapy group compared that in the other two groups. The tumor cells showed different degrees of apoptosis in the thermochemotherapy and chemotherapy groups under electron microscope.Conclusion: The tissue specific CD/5FC thermochemotherapy system can inhibit the growth of liver metastasis of colon cancer in nude mice.
2009, 16(4):391-395.
DOI: 10.3872/j.issn.1007-385X.2009.4.015
Abstract:
Objective:To prepare and identify monoclonal antibody specifically targeting epidermal growth factor receptor (EGFR) and (or) epidermal growth factor receptor vⅢ(EGFRvⅢ), and to investigate its inhibitory effects on human hepatocellular carcinoma Huh7EGFRvⅢ cell and epidermal carcinoma A431 cellimplanted tumors in nude mice. Methods:BALB/c mice were immunized with 3T3 cells stably transfected with EGFRvⅢ (3T3EGFRvⅢ). Immunized spleen cells were fused with myeloma SP2/0 cells, and antiEGFRvⅢ monoclonal antibody positive hybridoma cells (named 9B9 antibody and 9B9 cells, respectively) were selected and identified by ELISA. The specific interaction between 9B9 antibody and EGFRvⅢ/EGFR antigen was detected by Western blotting and immunofluorescence assay. Huh7EGFRvⅢ cell(human hepatocellular carcinoma Huh7 cells stably transfected with EGFRvⅢ) and epidermal cell carcinoma A431 cellbearing mouse models were established and were divided into PBS group, Cetuximab group and 9B9 antibody group. Then, antitumor effect of 9B9 antibody was examined and compared with those of PBS and Cetuximab. Results:A monoclonal antibody, named 9B9 antibody, was obtained by hybridoma technique and it reacted with both EGFRvⅢ antigen and EGFR antigen as detected by Western blotting and immunofluorescence. The inhibitory rates of Cetuximab and 9B9 antibody against Huh7EGFRvⅢ cellsimplanted tumors were 42% and 46%, respectively, and those against A431 cellsimplanted tumors were 85% and 86%, respectively. Conclusion:9B9 monoclonal antibody can effectively inhibit the growth of human hepatocellular carcinoma cell and epidermal cell carcinoma cellimplanted tumors, and the effects resemble that of Cetuximab.
Objective:To prepare and identify monoclonal antibody specifically targeting epidermal growth factor receptor (EGFR) and (or) epidermal growth factor receptor vⅢ(EGFRvⅢ), and to investigate its inhibitory effects on human hepatocellular carcinoma Huh7EGFRvⅢ cell and epidermal carcinoma A431 cellimplanted tumors in nude mice. Methods:BALB/c mice were immunized with 3T3 cells stably transfected with EGFRvⅢ (3T3EGFRvⅢ). Immunized spleen cells were fused with myeloma SP2/0 cells, and antiEGFRvⅢ monoclonal antibody positive hybridoma cells (named 9B9 antibody and 9B9 cells, respectively) were selected and identified by ELISA. The specific interaction between 9B9 antibody and EGFRvⅢ/EGFR antigen was detected by Western blotting and immunofluorescence assay. Huh7EGFRvⅢ cell(human hepatocellular carcinoma Huh7 cells stably transfected with EGFRvⅢ) and epidermal cell carcinoma A431 cellbearing mouse models were established and were divided into PBS group, Cetuximab group and 9B9 antibody group. Then, antitumor effect of 9B9 antibody was examined and compared with those of PBS and Cetuximab. Results:A monoclonal antibody, named 9B9 antibody, was obtained by hybridoma technique and it reacted with both EGFRvⅢ antigen and EGFR antigen as detected by Western blotting and immunofluorescence. The inhibitory rates of Cetuximab and 9B9 antibody against Huh7EGFRvⅢ cellsimplanted tumors were 42% and 46%, respectively, and those against A431 cellsimplanted tumors were 85% and 86%, respectively. Conclusion:9B9 monoclonal antibody can effectively inhibit the growth of human hepatocellular carcinoma cell and epidermal cell carcinoma cellimplanted tumors, and the effects resemble that of Cetuximab.
2009, 16(4):396-400.
DOI: 10.3872/j.issn.1007-385X.2009.4.016
Abstract:
Objective:To study the therapeutic effect of local implantation with temozolomide/PLGA microsphere (TMMS) on rat C6 glioma in vivo.Methods: C6 glioma cells were implanted stereotaxically to rat caudate nucleus of left cerebrum to establish C6 gliomarat model. C6 gliomarats were treated with oral temozolomide or with TMMS (implanted in cerebral tumor foci). The general manifestation, survival time, tumor size and pathological changes were observed in each group. The expression of proliferation cell nuclear antigen (PCNA) in glioma tissues was examined by immunohistochemistry method. Apoptosis of glioma cells was measured by TUNEL.Results: The survival period of C6 gliomarats in TMMS group was longer than those in the sham group, blank microsphere group and oral Temozolomide group (being \[31.2±6.21\] vs \[20.7±4.83\], \[19.2±6.23\] and \[24.7±6.31\] d, respectively, P<0.05 or P<0.01). MRI results demonstrated that the volume of glioma in interstitial TMMS group was smaller than those in the sham group, blank microsphere group and oral Temozolomide group (being \[28.8±6.41\] vs \[56.4±6.92\], \[58.2±5.36\] and \[46.7±7.28\] mm3, respectively, P<0.05 or P<0.01). PCNA expression in glioma tissues of TMMS group was significantly lower compared with those in the sham group, blank microshere group and oral Temozolomide group (being \[202±433\]% vs \[63.2±5.91\]%, \[62.1±7.88\]% and \[41.7±6.71\]%, respectively, P<0.01). Apoptosis rate of glioma cells in TMMS group was markedly higher compared with those in the sham group, blank microshere group and oral Temozolomide group (being \[32.31±3.17\]% vs \[8.63±1.52\]%, \[9.25±2.31\]% and \[16.14±3.42\]%, respectively, P<001).Conclusion: Interstitial TMMS therapy effectively inhibits proliferation and induces apoptosis of glioma cells in mice, and it has a potential in clinic tumor therapy.
Objective:To study the therapeutic effect of local implantation with temozolomide/PLGA microsphere (TMMS) on rat C6 glioma in vivo.Methods: C6 glioma cells were implanted stereotaxically to rat caudate nucleus of left cerebrum to establish C6 gliomarat model. C6 gliomarats were treated with oral temozolomide or with TMMS (implanted in cerebral tumor foci). The general manifestation, survival time, tumor size and pathological changes were observed in each group. The expression of proliferation cell nuclear antigen (PCNA) in glioma tissues was examined by immunohistochemistry method. Apoptosis of glioma cells was measured by TUNEL.Results: The survival period of C6 gliomarats in TMMS group was longer than those in the sham group, blank microsphere group and oral Temozolomide group (being \[31.2±6.21\] vs \[20.7±4.83\], \[19.2±6.23\] and \[24.7±6.31\] d, respectively, P<0.05 or P<0.01). MRI results demonstrated that the volume of glioma in interstitial TMMS group was smaller than those in the sham group, blank microsphere group and oral Temozolomide group (being \[28.8±6.41\] vs \[56.4±6.92\], \[58.2±5.36\] and \[46.7±7.28\] mm3, respectively, P<0.05 or P<0.01). PCNA expression in glioma tissues of TMMS group was significantly lower compared with those in the sham group, blank microshere group and oral Temozolomide group (being \[202±433\]% vs \[63.2±5.91\]%, \[62.1±7.88\]% and \[41.7±6.71\]%, respectively, P<0.01). Apoptosis rate of glioma cells in TMMS group was markedly higher compared with those in the sham group, blank microshere group and oral Temozolomide group (being \[32.31±3.17\]% vs \[8.63±1.52\]%, \[9.25±2.31\]% and \[16.14±3.42\]%, respectively, P<001).Conclusion: Interstitial TMMS therapy effectively inhibits proliferation and induces apoptosis of glioma cells in mice, and it has a potential in clinic tumor therapy.
2009, 16(4):401-404.
DOI: 10.3872/j.issn.1007-385X.2009.4.017
Abstract:
Objective:To investigate CD1a and CD83 expression in tumor infiltrating dendritic cell (TIDC) of esophageal squamous cell carcinoma (ESCC) and tumorinfiltrating lymph node tissues, and to discuss its relationship with the clinical pathological characteristics of esophageal carcinoma. Methods:Seventyeight paraffin samples were obtained from pathologically diagnosed esophageal carcinoma patients in the fourth hospital of Hebei medical university during 2002 to 2003. Flow cytometry was used to examine the expression of CD1a and CD83 on TIDC in 78 esophageal carcinoma tissues, 24 normal esophageal mucosal samples, 35 normal lymph nodes and 32 tumorinfiltrating lymph nodes.Results: (1) The expression of CD1a and CD83 in ESCC tissues was significantly lower than those in the normal esophageal mucosa (P<005). (2) CD1a expression was not correlated with the tumor infiltration, clinical stage and lymph node metastasis of esophageal carcinoma (all P>0.05), while CD83 expression was correlated with the clinical stage and lymph node metastasis (all P<0.05). (3) There was no significant difference in CD1a expression between normal lymph node and tumorinfiltrating lymph node (P>0.05), but CD83 expression in tumorinfiltrating lymph node was less than that in the normal lymph node (P<005). Conclusion：The expression of CD83 in TIDC can reflect the local immune status of esophageal carcinoma, and also plays an important role in the development and progression of esophageal carcinoma. CD83 expression may serve as a biomarker for evaluating biological characteristics of esophageal carcinoma.
Objective:To investigate CD1a and CD83 expression in tumor infiltrating dendritic cell (TIDC) of esophageal squamous cell carcinoma (ESCC) and tumorinfiltrating lymph node tissues, and to discuss its relationship with the clinical pathological characteristics of esophageal carcinoma. Methods:Seventyeight paraffin samples were obtained from pathologically diagnosed esophageal carcinoma patients in the fourth hospital of Hebei medical university during 2002 to 2003. Flow cytometry was used to examine the expression of CD1a and CD83 on TIDC in 78 esophageal carcinoma tissues, 24 normal esophageal mucosal samples, 35 normal lymph nodes and 32 tumorinfiltrating lymph nodes.Results: (1) The expression of CD1a and CD83 in ESCC tissues was significantly lower than those in the normal esophageal mucosa (P<005). (2) CD1a expression was not correlated with the tumor infiltration, clinical stage and lymph node metastasis of esophageal carcinoma (all P>0.05), while CD83 expression was correlated with the clinical stage and lymph node metastasis (all P<0.05). (3) There was no significant difference in CD1a expression between normal lymph node and tumorinfiltrating lymph node (P>0.05), but CD83 expression in tumorinfiltrating lymph node was less than that in the normal lymph node (P<005). Conclusion：The expression of CD83 in TIDC can reflect the local immune status of esophageal carcinoma, and also plays an important role in the development and progression of esophageal carcinoma. CD83 expression may serve as a biomarker for evaluating biological characteristics of esophageal carcinoma.
2009, 16(4):405-409.
DOI: 10.3872/j.issn.1007-385X.2009.4.018
Abstract:
Objective: To study HER2 levels in the serum and breast cancer tissues and their correlation with clinical parameters, so as to explore drugs selection and prognosis prediction of breast cancer. Methods: Sixtyseven pathologicallyconfirmed breast cancer patients, 20 patients with breast benign tumor, and 20 healthy women, who were treated in Fujian tumor hospital from Jan. 2008 to Oct. 2008 were included in this study. Expression of HER2 in breast cancer tissues was examined by immunohistochemistry, and serum HER2 level in breast cancer patients was examined by ELISA. Results: Serum level and positive rate of serum HER2 in breast cancer patients were significantly higher than those in healthy women and breast benign tumor patients (P<0.05). Serum HER level and positive rate of serum HER2 in histological HER2 positive breast cancer patients were significantly higher than those in histological HER2 negative patients (P<0.05), with the positive rate of serum HER positively correlated with histological HER2 status. The result of serological method was consisted well with the histological method in the determination of HER2 status in breast cancer patients. Serum HER2 level in some histological HER2 negative primary breast cancer patients was increased after recurrence and metastasis, and positive rate of serum HER2 in Ⅳ metastatic breast cancer patients was significantly higher than that in ⅠⅢ metastatic breast cancer patients (P<0.05). Conclusions: Serum HER2 level is elevated in breast cancer patients, and positive rate of serum HER2 is correlated with histological HER2 status and clinical stages of cancer. Determination of serum HER2 may be useful in diagnosis, HER2 status evaluation and prognosis prediction of breast cancer patients.
Objective: To study HER2 levels in the serum and breast cancer tissues and their correlation with clinical parameters, so as to explore drugs selection and prognosis prediction of breast cancer. Methods: Sixtyseven pathologicallyconfirmed breast cancer patients, 20 patients with breast benign tumor, and 20 healthy women, who were treated in Fujian tumor hospital from Jan. 2008 to Oct. 2008 were included in this study. Expression of HER2 in breast cancer tissues was examined by immunohistochemistry, and serum HER2 level in breast cancer patients was examined by ELISA. Results: Serum level and positive rate of serum HER2 in breast cancer patients were significantly higher than those in healthy women and breast benign tumor patients (P<0.05). Serum HER level and positive rate of serum HER2 in histological HER2 positive breast cancer patients were significantly higher than those in histological HER2 negative patients (P<0.05), with the positive rate of serum HER positively correlated with histological HER2 status. The result of serological method was consisted well with the histological method in the determination of HER2 status in breast cancer patients. Serum HER2 level in some histological HER2 negative primary breast cancer patients was increased after recurrence and metastasis, and positive rate of serum HER2 in Ⅳ metastatic breast cancer patients was significantly higher than that in ⅠⅢ metastatic breast cancer patients (P<0.05). Conclusions: Serum HER2 level is elevated in breast cancer patients, and positive rate of serum HER2 is correlated with histological HER2 status and clinical stages of cancer. Determination of serum HER2 may be useful in diagnosis, HER2 status evaluation and prognosis prediction of breast cancer patients.
2009, 16(4):410-412.
DOI: 10.3872/j.issn.1007-385X.2009.4.019
Abstract:
目的：观察重组改构人肿瘤坏死因子（recombined mutant human tumor necrosis factor, rmhTNF）治疗恶性腹腔积液的疗效，并分析其影响因素。方法：回顾分析2004年至2007年在军区总医院肿瘤科住院并行腹腔置管灌注rmhTNF治疗的96例肿瘤(结肠癌34例、卵巢癌17例、胃癌16例、肝癌29例)腹腔积液患者的临床资料，通过对患者年龄、性别、用药剂量、组织来源、积液量等因素的治疗有效率分析，判断这些因素对疗效的影响。结果：96例患者中，显效和有效的病例共70例(72.9%)。在单因素分析中，rmhTNF疗效在患者性别、年龄等方面差异不明显(P>0.05)，而对于具有大量积液、KPS评分<60分、 TNF小剂量(500万U)、肝癌患者的治疗效果较差(P<0.05或P<0.01)；多因素分析显示，用药前的积液量、给药剂量是影响疗效的重要因素。全部受治患者都无明显的毒性反应。结论：rmhTNF治疗恶性腹腔积液疗效较好，尤其是对一般情况较好的中少量积液患者采用大剂量治疗的效果更明显。
目的：观察重组改构人肿瘤坏死因子（recombined mutant human tumor necrosis factor, rmhTNF）治疗恶性腹腔积液的疗效，并分析其影响因素。方法：回顾分析2004年至2007年在军区总医院肿瘤科住院并行腹腔置管灌注rmhTNF治疗的96例肿瘤(结肠癌34例、卵巢癌17例、胃癌16例、肝癌29例)腹腔积液患者的临床资料，通过对患者年龄、性别、用药剂量、组织来源、积液量等因素的治疗有效率分析，判断这些因素对疗效的影响。结果：96例患者中，显效和有效的病例共70例(72.9%)。在单因素分析中，rmhTNF疗效在患者性别、年龄等方面差异不明显(P>0.05)，而对于具有大量积液、KPS评分<60分、 TNF小剂量(500万U)、肝癌患者的治疗效果较差(P<0.05或P<0.01)；多因素分析显示，用药前的积液量、给药剂量是影响疗效的重要因素。全部受治患者都无明显的毒性反应。结论：rmhTNF治疗恶性腹腔积液疗效较好，尤其是对一般情况较好的中少量积液患者采用大剂量治疗的效果更明显。
2009, 16(4):413-417.
DOI: 10.3872/j.issn.1007-385X.2009.4.020
Abstract:
基因调控、信号转导通路异常均可引起细胞增殖失控，导致肿瘤发生。肿瘤细胞对化疗药物耐药是肿瘤患者死亡的主要原因。细胞内药物有效浓度的降低、DNA损伤的修复障碍、基因的突变及异常表达、信号转导通路的异常等均参与了肿瘤细胞的多药耐药。张力蛋白同源10号染色体缺失的磷酸酶基因（phosphatase and tension homology deleted on chromosome ten gene，PTEN）是具有磷酸酶活性的抑癌基因，在多种肿瘤细胞中异常表达，主要通过抑制PI3K/Akt/mTOR（mammalian target of rapamycin, mTOR）等多种信号转导通路参与细胞的增殖、凋亡及化疗耐药。因此，上调野生型PTEN的表达，或使用PI3K/Akt/mTOR信号通路抑制剂，可逆转肿瘤细胞的多药耐药，提高传统化疗的疗效。
基因调控、信号转导通路异常均可引起细胞增殖失控，导致肿瘤发生。肿瘤细胞对化疗药物耐药是肿瘤患者死亡的主要原因。细胞内药物有效浓度的降低、DNA损伤的修复障碍、基因的突变及异常表达、信号转导通路的异常等均参与了肿瘤细胞的多药耐药。张力蛋白同源10号染色体缺失的磷酸酶基因（phosphatase and tension homology deleted on chromosome ten gene，PTEN）是具有磷酸酶活性的抑癌基因，在多种肿瘤细胞中异常表达，主要通过抑制PI3K/Akt/mTOR（mammalian target of rapamycin, mTOR）等多种信号转导通路参与细胞的增殖、凋亡及化疗耐药。因此，上调野生型PTEN的表达，或使用PI3K/Akt/mTOR信号通路抑制剂，可逆转肿瘤细胞的多药耐药，提高传统化疗的疗效。
2009, 16(4):418-421.
DOI: 10.3872/j.issn.1007-385X.2009.4.021
Abstract:
MicroRNAs (miRNAs)是一类内源性、非编码的单链小分子RNA，作用广泛，参与生命活动中的一系列重要进程，并与肿瘤的发生、发展密切相关。miRNA let7是最早发现的miRNA之一，是线虫时序性发育的关键性调控因子；在哺乳动物中调节多种细胞增殖，且在细胞周期调节中起关键作用。let-7与人类多种癌症的发生、发展有关，其中与肺癌的关系最为密切；hsalet7在肺癌中表达显著降低，在非小细胞肺癌（NSCLC）中尤为多见，其低表达可能与肿瘤预后不良有关，而高表达则直接抑制肺癌生长；let-7作为肿瘤抑制因子负性调控多种癌基因，如RAS、高迁移率蛋白A2基因(high mobility group protein A2，HMGA2)等；同时也负性调控多种细胞周期调节因子，如CDC25A、CDK6、Cyclin D2。let7在肺癌组织中起到了肿瘤抑制因子的作用，有望成为肺癌基因治疗和预后判断的一个新靶标。
MicroRNAs (miRNAs)是一类内源性、非编码的单链小分子RNA，作用广泛，参与生命活动中的一系列重要进程，并与肿瘤的发生、发展密切相关。miRNA let7是最早发现的miRNA之一，是线虫时序性发育的关键性调控因子；在哺乳动物中调节多种细胞增殖，且在细胞周期调节中起关键作用。let-7与人类多种癌症的发生、发展有关，其中与肺癌的关系最为密切；hsalet7在肺癌中表达显著降低，在非小细胞肺癌（NSCLC）中尤为多见，其低表达可能与肿瘤预后不良有关，而高表达则直接抑制肺癌生长；let-7作为肿瘤抑制因子负性调控多种癌基因，如RAS、高迁移率蛋白A2基因(high mobility group protein A2，HMGA2)等；同时也负性调控多种细胞周期调节因子，如CDC25A、CDK6、Cyclin D2。let7在肺癌组织中起到了肿瘤抑制因子的作用，有望成为肺癌基因治疗和预后判断的一个新靶标。
2009, 16(4):422-426.
DOI: 10.3872/j.issn.1007-385X.2009.4.022
Abstract:
骨转移是乳腺癌、前列腺癌等晚期恶性肿瘤的常见并发症。癌细胞增殖转移到骨引起溶骨性和成骨性骨损伤，其发生是多个因素共同作用的结果。近年来的研究发现肿瘤细胞与骨微环境之间存在相互作用，骨基质中富含的某些细胞因子如转化生长因子(TGFβ)、胰岛素样生长因子(IGFⅠ和IGFⅡ)等直接促进肿瘤细胞生长，并在维持骨形成和骨破坏的动态平衡中发挥重要作用。骨微环境中的物理因素包括缺氧、高钙、酸中毒等也为肿瘤生长提供适宜的条件。本文主要从分子水平阐述肿瘤骨转移过程中涉及到的肿瘤细胞与骨微环境之间的相互作用。
骨转移是乳腺癌、前列腺癌等晚期恶性肿瘤的常见并发症。癌细胞增殖转移到骨引起溶骨性和成骨性骨损伤，其发生是多个因素共同作用的结果。近年来的研究发现肿瘤细胞与骨微环境之间存在相互作用，骨基质中富含的某些细胞因子如转化生长因子(TGFβ)、胰岛素样生长因子(IGFⅠ和IGFⅡ)等直接促进肿瘤细胞生长，并在维持骨形成和骨破坏的动态平衡中发挥重要作用。骨微环境中的物理因素包括缺氧、高钙、酸中毒等也为肿瘤生长提供适宜的条件。本文主要从分子水平阐述肿瘤骨转移过程中涉及到的肿瘤细胞与骨微环境之间的相互作用。
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- 《中国肿瘤生物治疗杂志》
- 1994年创刊
- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
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- 刊号：ISSN 1007-385X
- CN 31-1725/R
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.