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Volume 16,Issue 3,2009 Table of Contents
2009, 16(3):211-215.
DOI: 10.3872/j.issn.1007-385X.2009.3.001
Abstract:
Dendritic cells (DCs) have different subtypes with distinct phenotypes and biological functions. Meloyiod dendritic cell subtype is one of the most important DCs subtypes. Recent studies have revealed that human monocytes were composed of CD14++CD16- and CD14+CD16+ subtypes, and mouse monocytes consisted of CD115+Ly6Chigh and CD115+Ly6Clow/ subtypes. Different monocyte subsets differentiate into different dendritic cells subsets with distinct phenotypes and induce different types of immune responses in vivo or in vitro. Under steady state, mouse CD115+Ly6Clow monocyte subtype is an important precursor of dendritic cells in peripheral organs and tissues, but in inflammatory response, CD115+Ly6Chigh monocyte subtype is the main precursor of dendritic cells in peripheral lymphoid organs. CD115+Ly6Chigh monocyte derived dendritic cells can directly present antigens that acquired in peripheral tissues after differentiating into dendritic cells, and transfer their MHCⅠ/peptide complex to residential dendritic cells as well. The cooperation and interaction of dendritic cells from different sources enable the immune system to respond to different stimuli properly.
Dendritic cells (DCs) have different subtypes with distinct phenotypes and biological functions. Meloyiod dendritic cell subtype is one of the most important DCs subtypes. Recent studies have revealed that human monocytes were composed of CD14++CD16- and CD14+CD16+ subtypes, and mouse monocytes consisted of CD115+Ly6Chigh and CD115+Ly6Clow/ subtypes. Different monocyte subsets differentiate into different dendritic cells subsets with distinct phenotypes and induce different types of immune responses in vivo or in vitro. Under steady state, mouse CD115+Ly6Clow monocyte subtype is an important precursor of dendritic cells in peripheral organs and tissues, but in inflammatory response, CD115+Ly6Chigh monocyte subtype is the main precursor of dendritic cells in peripheral lymphoid organs. CD115+Ly6Chigh monocyte derived dendritic cells can directly present antigens that acquired in peripheral tissues after differentiating into dendritic cells, and transfer their MHCⅠ/peptide complex to residential dendritic cells as well. The cooperation and interaction of dendritic cells from different sources enable the immune system to respond to different stimuli properly.
2009, 16(3):216-226.
DOI: 10.3872/j.issn.1007-385X.2009.3.002
Abstract:
Objective:To investigate the arginine (Arg) sites in splicing factor 2/alternative splicing factor (SF2/ASF) methylated by protein arginine methyltransferase 1 (PRMT1). Methods: Wildtype and Arg93, Arg97, Arg109 mutant SF2/ASF plasmids were constructed, and GSTPRMT1, GSTSF2/ASF and arginine mutant GSTSF2/ASF fusion proteins were induced and purified. Methylation activity of PRMT1 on wildtype or mutant SF2/ASF protein and methylated sites of SF2/ASF were examined by methylation assay. The effect of SF2/ASF methylation on its subcellular localization was analyzed by immunofluorescence assay.Results: PRMT1 induced methylation of SF2/ASF at arginine, and PRMT1 did not methylate SF2/ASF when SF2/ASF was mutant at Arg93, Arg97 or Arg109, with Arg97 mutation showing the most profound inhibitory effect. Methylation of SF2/ASF did not affect its subcellular localization.Conclusion: SF2/ASF is a newly identified substrate of PRMT1; Arg93, Arg97 and Arg 109 are the three methylation sites in SF2/ASF, and Arg97 is the main methylation site. Methylation of SF2/ASF does not affect its subcellular localization.
Objective:To investigate the arginine (Arg) sites in splicing factor 2/alternative splicing factor (SF2/ASF) methylated by protein arginine methyltransferase 1 (PRMT1). Methods: Wildtype and Arg93, Arg97, Arg109 mutant SF2/ASF plasmids were constructed, and GSTPRMT1, GSTSF2/ASF and arginine mutant GSTSF2/ASF fusion proteins were induced and purified. Methylation activity of PRMT1 on wildtype or mutant SF2/ASF protein and methylated sites of SF2/ASF were examined by methylation assay. The effect of SF2/ASF methylation on its subcellular localization was analyzed by immunofluorescence assay.Results: PRMT1 induced methylation of SF2/ASF at arginine, and PRMT1 did not methylate SF2/ASF when SF2/ASF was mutant at Arg93, Arg97 or Arg109, with Arg97 mutation showing the most profound inhibitory effect. Methylation of SF2/ASF did not affect its subcellular localization.Conclusion: SF2/ASF is a newly identified substrate of PRMT1; Arg93, Arg97 and Arg 109 are the three methylation sites in SF2/ASF, and Arg97 is the main methylation site. Methylation of SF2/ASF does not affect its subcellular localization.
2009, 16(3):221-231.
DOI: 10.3872/j.issn.1007-385X.2009.3.003
Abstract:
Objective:To determine whether the influenza vaccine can affect the function of dendritic cells（DCs）derived from the bone marrow of patients with myeloid leukemia and the possible mechanism. Methods: The bone marrow (BM) mononuclear cells were obtained from 19 patients with acute myelocytic leukemia (AML) and 8 patients with chronic myeloid leukemia (CML), and were cultured with GMCSF and IL4 for 7 days to obtain immature DCs. Then DCs were stimulated by whole inactivated influenza vaccine (WIV), split influenza vaccine (SIV), or TNFα. After 24 h, phenotypes and karyotypes of these DCs were assessed by FACS and R band karyotype analysis, respectively. The supernatant IL12 levels were measured by ELISA in each group. Cytotoxic activity of CTL induced by differently treated DCs was measured by CCK8 assay.Results: DCs were successfully induced in 15 of the 19 AML patients and all the 8 CML patients. After stimulated with WIV or SIV for 24 h, DCs exhibited enhanced expression of CD83, CD86 and HLADR, and increased secretion of IL12 (all P<0.05). CTL induced by WIV or SIVstimulated DCs specifically killed autologous leukemia cells in vitro (P<0.05). Furthermore, WIVstimulated DCs were more powerful than SIVstimulated DCs in killing target cells (P<0.05). Conclusion: Influenza vaccine can promote the maturation and IL12 secretion of DCs derived from myeloid leukemia patients, and CTL induced by influenza vaccinestimulated DCs has a stronger ability to kill autologous leukemia cells.
Objective:To determine whether the influenza vaccine can affect the function of dendritic cells（DCs）derived from the bone marrow of patients with myeloid leukemia and the possible mechanism. Methods: The bone marrow (BM) mononuclear cells were obtained from 19 patients with acute myelocytic leukemia (AML) and 8 patients with chronic myeloid leukemia (CML), and were cultured with GMCSF and IL4 for 7 days to obtain immature DCs. Then DCs were stimulated by whole inactivated influenza vaccine (WIV), split influenza vaccine (SIV), or TNFα. After 24 h, phenotypes and karyotypes of these DCs were assessed by FACS and R band karyotype analysis, respectively. The supernatant IL12 levels were measured by ELISA in each group. Cytotoxic activity of CTL induced by differently treated DCs was measured by CCK8 assay.Results: DCs were successfully induced in 15 of the 19 AML patients and all the 8 CML patients. After stimulated with WIV or SIV for 24 h, DCs exhibited enhanced expression of CD83, CD86 and HLADR, and increased secretion of IL12 (all P<0.05). CTL induced by WIV or SIVstimulated DCs specifically killed autologous leukemia cells in vitro (P<0.05). Furthermore, WIVstimulated DCs were more powerful than SIVstimulated DCs in killing target cells (P<0.05). Conclusion: Influenza vaccine can promote the maturation and IL12 secretion of DCs derived from myeloid leukemia patients, and CTL induced by influenza vaccinestimulated DCs has a stronger ability to kill autologous leukemia cells.
2009, 16(3):227-232.
DOI: 10.3872/j.issn.1007-385X.2009.3.004
Abstract:
Objective: To predict and identify survivin specific HLAA2+ CTL restricted epitopes by bioinformatic methods, so as to provide a foundation for survivinbased immunotherapy. Methods: Survivin specific HLAA2+ restricted CTL epotides were predicted by computer supermotif algorithm combined with quantitativemotif algorithm. Candidate epitopes were verified when their scores were higher than 10 and were then artificially synthesized. Affinity of candidate epitope was examined by HLAA2 binding assay combined with flow cytometry using T2 cells (shown as fluorescence index, FI). Stability of candidate epitope was evaluated by HLAA2 dissociation assay combined with flow cytometry (shown as 50% complex dissociation time, DC50). Results: Nine candidate epotides were obtained: 20STFKNWPFL28 (SV2028), 23KNWPFLEGC31 (SV2331), 96LTLGEFLKL104 (SV96104), 6LPPAWQPFL14 (SV614), 33CTPERMAEA41 (SV3341), 46CPTENEPDL54 (SV4654), 130KVRRAIEQL138(SV130138), 37RMAEAGFIH45(SV3745), and 88SVKKQFEEL96 (SV8896). HLAA2 binding assay showed that FI values of SV2028, SV96104, SV130138 and SV2331 epotides were 8.61, 688, 5.89 and 3.81, respectively; those of SV3341, SV614, SV4654, SV3745 and SV8896 epotides were 0.31, -0.29,-04,-0.16 and -0.03, respectively. HLAA2 dissociation assay showed that DC50 values of SV2028, SV96104 and SV130138 epotides were longer than 8 h; that of SV2331 epotide was 46 h; those of SV614, SV3341and SV8896 epotides were all 24 h; those of SV4654, SV3745 epotides were both 02 h. The above results demonstrated that SV2028, SV96104 and SV130138 were high affinity epotides; SV2331 was intermediate affinity epotide; and SV3341, SV614, SV4654, SV3745 and SV8896 were low affinity epotides. Conclusion: Antigen epitope can be quickly and efficiently predicted by supermotif algorithm combined with quantitativemotif algorithm. SV2028, SV96104 and SV130138 epitopes are survivin specific HLAA2+ restricted CTL epotides, which can be used for later research.
Objective: To predict and identify survivin specific HLAA2+ CTL restricted epitopes by bioinformatic methods, so as to provide a foundation for survivinbased immunotherapy. Methods: Survivin specific HLAA2+ restricted CTL epotides were predicted by computer supermotif algorithm combined with quantitativemotif algorithm. Candidate epitopes were verified when their scores were higher than 10 and were then artificially synthesized. Affinity of candidate epitope was examined by HLAA2 binding assay combined with flow cytometry using T2 cells (shown as fluorescence index, FI). Stability of candidate epitope was evaluated by HLAA2 dissociation assay combined with flow cytometry (shown as 50% complex dissociation time, DC50). Results: Nine candidate epotides were obtained: 20STFKNWPFL28 (SV2028), 23KNWPFLEGC31 (SV2331), 96LTLGEFLKL104 (SV96104), 6LPPAWQPFL14 (SV614), 33CTPERMAEA41 (SV3341), 46CPTENEPDL54 (SV4654), 130KVRRAIEQL138(SV130138), 37RMAEAGFIH45(SV3745), and 88SVKKQFEEL96 (SV8896). HLAA2 binding assay showed that FI values of SV2028, SV96104, SV130138 and SV2331 epotides were 8.61, 688, 5.89 and 3.81, respectively; those of SV3341, SV614, SV4654, SV3745 and SV8896 epotides were 0.31, -0.29,-04,-0.16 and -0.03, respectively. HLAA2 dissociation assay showed that DC50 values of SV2028, SV96104 and SV130138 epotides were longer than 8 h; that of SV2331 epotide was 46 h; those of SV614, SV3341and SV8896 epotides were all 24 h; those of SV4654, SV3745 epotides were both 02 h. The above results demonstrated that SV2028, SV96104 and SV130138 were high affinity epotides; SV2331 was intermediate affinity epotide; and SV3341, SV614, SV4654, SV3745 and SV8896 were low affinity epotides. Conclusion: Antigen epitope can be quickly and efficiently predicted by supermotif algorithm combined with quantitativemotif algorithm. SV2028, SV96104 and SV130138 epitopes are survivin specific HLAA2+ restricted CTL epotides, which can be used for later research.
2009, 16(3):232-237.
DOI: 10.3872/j.issn.1007-385X.2009.3.005
Abstract:
Objective:To compare the effects of antisense (AS) bcl2, interleukin6（IL6） and interleukin6 receptor phosphorothioate oligodeoxynucleotides (PSODN) on the proliferation and apoptosis of smallcell lung cancer NCIH446 cells, and to explore strategies of target genes selection in antisense oligodeoxynucleotides therapy. Methods:Bcl2 ASPSODN, IL6 ASPSODN and IL6R ASPSODN were synthesized and were used to treat NCIH446 cells. Proliferation and viability of NCIH446 cells were measured by cel1 clone formation assay after treatment; apoptosis of NCIH446 cells was detected by DNA content analysis and MitoCapture detection kit; and the expression of bcl2, IL6 and IL6R mRNA was examined by RTPCR. Results: （1）All the three antisense phosphorothioate oligodeoxynucleotides IL6, bcl2, IL6R ASPSODN inhibited proliferation and viability of lung cancer NCIH446 cells, and induced NCIH446 cells apoptosis, with the inhibitory effect of IL6 ASPSODN more effective than those of bcl2 ASPSODN and IL6R ASPSODN. （2）All the three kinds of ASPSODN decreased the corresponding gene expression, with gene expression decreased by (84.1±5.01)%, (62.6±3.42)% and (60.3±4.45)% in IL6, bcl2 and IL6R ASPSODN group, respectively. （3）ASPSODN not only inhibited the expression of its target gene, but also regulated expression of other genes. IL6 ASPSODN decreased bcl2 expression by (32.2±0.20)% and bcl2 ASPSODN increased IL6 expression by (74.3±413)%. Conclusion: IL6 ASPSODN is more effective than bcl2 and IL6R ASPSODN in inducing apoptosis of smallcell lung cancer NCIH446 cells, so IL6 may be used as a target gene in antisense oligodeoxynucleotide therapy of smallcell lung cancer.
Objective:To compare the effects of antisense (AS) bcl2, interleukin6（IL6） and interleukin6 receptor phosphorothioate oligodeoxynucleotides (PSODN) on the proliferation and apoptosis of smallcell lung cancer NCIH446 cells, and to explore strategies of target genes selection in antisense oligodeoxynucleotides therapy. Methods:Bcl2 ASPSODN, IL6 ASPSODN and IL6R ASPSODN were synthesized and were used to treat NCIH446 cells. Proliferation and viability of NCIH446 cells were measured by cel1 clone formation assay after treatment; apoptosis of NCIH446 cells was detected by DNA content analysis and MitoCapture detection kit; and the expression of bcl2, IL6 and IL6R mRNA was examined by RTPCR. Results: （1）All the three antisense phosphorothioate oligodeoxynucleotides IL6, bcl2, IL6R ASPSODN inhibited proliferation and viability of lung cancer NCIH446 cells, and induced NCIH446 cells apoptosis, with the inhibitory effect of IL6 ASPSODN more effective than those of bcl2 ASPSODN and IL6R ASPSODN. （2）All the three kinds of ASPSODN decreased the corresponding gene expression, with gene expression decreased by (84.1±5.01)%, (62.6±3.42)% and (60.3±4.45)% in IL6, bcl2 and IL6R ASPSODN group, respectively. （3）ASPSODN not only inhibited the expression of its target gene, but also regulated expression of other genes. IL6 ASPSODN decreased bcl2 expression by (32.2±0.20)% and bcl2 ASPSODN increased IL6 expression by (74.3±413)%. Conclusion: IL6 ASPSODN is more effective than bcl2 and IL6R ASPSODN in inducing apoptosis of smallcell lung cancer NCIH446 cells, so IL6 may be used as a target gene in antisense oligodeoxynucleotide therapy of smallcell lung cancer.
2009, 16(3):238-242.
DOI: 10.3872/j.issn.1007-385X.2009.3.006
Abstract:
Objective:To study the inhibitory effect of a new antiATPase F1α antibody, antihuman angiostatin interacting and tumor metastasis involving protein (HAITMIP) antibody (MAb3D5AB1,GX), against lung adenocarcinoma cell line A549 in mouse tumor model. Methods: The tumorbearing mouse model was established by injecting A549 cells into the right infraaxillary dermis of C57BL/6 mice. Thirtytwo mice were evenly randomized into 4 groups. The control group was untreated; group A, B, and C were treated with different concentrations of GX (125, 250, 500 ng/ml, respectively). The tumor volume, tumor growth delay (TGD) and survival time of mice was observed in all groups. Microvascular density (MVD) of the tumors was determined by immunocytochemistry and apoptosis of tumor cells was examined by TUNEL assay. Results:Human lung adenocarcinoma A549 implanted animal model was successfully established in C57BL/6 mice. Tumor volumes in all GXtreated mice were smaller than that in control mice (P<0.05). TGD in all GXtreated mice was prolonged as the concentration of GX increasing. After treatment with GX, the MVD in tumors was significantly decreased (P<0.05) and survival time was increased (P<0.05) compared with those in control group. TUNEL results revealed that apoptosis rate of tumor cells of GXtreated mice was higher than that in control mice (P<001). Conclusion: GX can inhibit angiogenesis in implanted A549 tumors and promote apoptosis of A549 cells, thus inhibiting tumor growth and prolonging survival of tumorbearing mice.
Objective:To study the inhibitory effect of a new antiATPase F1α antibody, antihuman angiostatin interacting and tumor metastasis involving protein (HAITMIP) antibody (MAb3D5AB1,GX), against lung adenocarcinoma cell line A549 in mouse tumor model. Methods: The tumorbearing mouse model was established by injecting A549 cells into the right infraaxillary dermis of C57BL/6 mice. Thirtytwo mice were evenly randomized into 4 groups. The control group was untreated; group A, B, and C were treated with different concentrations of GX (125, 250, 500 ng/ml, respectively). The tumor volume, tumor growth delay (TGD) and survival time of mice was observed in all groups. Microvascular density (MVD) of the tumors was determined by immunocytochemistry and apoptosis of tumor cells was examined by TUNEL assay. Results:Human lung adenocarcinoma A549 implanted animal model was successfully established in C57BL/6 mice. Tumor volumes in all GXtreated mice were smaller than that in control mice (P<0.05). TGD in all GXtreated mice was prolonged as the concentration of GX increasing. After treatment with GX, the MVD in tumors was significantly decreased (P<0.05) and survival time was increased (P<0.05) compared with those in control group. TUNEL results revealed that apoptosis rate of tumor cells of GXtreated mice was higher than that in control mice (P<001). Conclusion: GX can inhibit angiogenesis in implanted A549 tumors and promote apoptosis of A549 cells, thus inhibiting tumor growth and prolonging survival of tumorbearing mice.
2009, 16(3):243-247.
DOI: 10.3872/j.issn.1007-385X.2009.3.007
Abstract:
Objective:To observe the effect of calcitonin generelated peptide (CGRP) on the expression of osteoprotegerin (OPG) and receptor activator of nuclear factorkappaB ligand (RANKL) in osteoblast cells through an in vitro breast cancer cell and osteoblast cell coculture system.Methods:The metastatic breast cancer MDAMB231 or MDAMB435 cells were cocultured with osteoblast MG63 cells to establish an in vitro microenvironment of bone metastasis of breast cancer. After treated with CGRP(1×108 mol/L), OPG and RANKL mRNA and protein expressions in osteoblast MG63 cells were examined by RTPCR and Western blotting. Results: Expression of RANKL in osteoblast MG63 cells was upregulated at both mRNA and protein levels when osteoblast MG63 cells were cocultured with breast cancer MDAMB231 or MDAMB435 cells, while those of OPG in osteoblast MG63 cells were both downregulated (P<0.05). After treatment with CGRP, expressions of RANKL in osteoblast MG63 cells were downregulated at both mRNA and protein levels, and the expressions of OPG mRNA and protein were both upregulated (P<0.05). Conclusion: Breast cancer MDAMB231 and MDAMB435 cells can promote osteolysis of osteoclast cells via regulating the expression of OPG/RANKL axis in osteoblast cells. CGRP can reverse the osteolysis of osteoblast cells induced by breast cancer cells and may serve as a potential therapeutic agent for treatment of bone metastasis of breast cancer.
Objective:To observe the effect of calcitonin generelated peptide (CGRP) on the expression of osteoprotegerin (OPG) and receptor activator of nuclear factorkappaB ligand (RANKL) in osteoblast cells through an in vitro breast cancer cell and osteoblast cell coculture system.Methods:The metastatic breast cancer MDAMB231 or MDAMB435 cells were cocultured with osteoblast MG63 cells to establish an in vitro microenvironment of bone metastasis of breast cancer. After treated with CGRP(1×108 mol/L), OPG and RANKL mRNA and protein expressions in osteoblast MG63 cells were examined by RTPCR and Western blotting. Results: Expression of RANKL in osteoblast MG63 cells was upregulated at both mRNA and protein levels when osteoblast MG63 cells were cocultured with breast cancer MDAMB231 or MDAMB435 cells, while those of OPG in osteoblast MG63 cells were both downregulated (P<0.05). After treatment with CGRP, expressions of RANKL in osteoblast MG63 cells were downregulated at both mRNA and protein levels, and the expressions of OPG mRNA and protein were both upregulated (P<0.05). Conclusion: Breast cancer MDAMB231 and MDAMB435 cells can promote osteolysis of osteoclast cells via regulating the expression of OPG/RANKL axis in osteoblast cells. CGRP can reverse the osteolysis of osteoblast cells induced by breast cancer cells and may serve as a potential therapeutic agent for treatment of bone metastasis of breast cancer.
2009, 16(3):248-252.
DOI: 10.3872/j.issn.1007-385X.2009.3.008
Abstract:
Objective:To construct pEGFPC1Smad4 expression vector and to observe the influence of Smad4 overexpression on the proliferation of human gastric cancer SGC7901 cells and its relationship with nuclear factor kappa B (NFκB). Methods:Recombinant expression vector pEGFPC1Smad4 was constructed and was used to transfect human gastric cancer SGC7901 cells. EGFP expression in transfected cells was detected by fluoroscopy. Smad4 and NFκB expression in transfectant was examined by Western blotting. Effect of Smad4 overexpression on activation of NFκB and proliferation of transfected SGC7901 cells were examined by electrophoretic mobility shift assay (EMSA) and MTT assay, respectively. Results: Expression of EGFP in transfected SGC7901 cells was observed under fluorescence microscope. Smad4 was overexpressed in transfected SGC7901 cells, accompanied by downregulation of NFκB p65 expression in the tranfectants. EMSA and MTT demonstrated that Smad4 overexpression significantly inhibited the activation of NFκB and the proliferation of SGC7901 cells (P<0.05, P<0.01).Conclusion: Smad4 overexpression can greatly inhibit the proliferation of human gastric cancer cells, probably through downregulation of NFκB pathway.
Objective:To construct pEGFPC1Smad4 expression vector and to observe the influence of Smad4 overexpression on the proliferation of human gastric cancer SGC7901 cells and its relationship with nuclear factor kappa B (NFκB). Methods:Recombinant expression vector pEGFPC1Smad4 was constructed and was used to transfect human gastric cancer SGC7901 cells. EGFP expression in transfected cells was detected by fluoroscopy. Smad4 and NFκB expression in transfectant was examined by Western blotting. Effect of Smad4 overexpression on activation of NFκB and proliferation of transfected SGC7901 cells were examined by electrophoretic mobility shift assay (EMSA) and MTT assay, respectively. Results: Expression of EGFP in transfected SGC7901 cells was observed under fluorescence microscope. Smad4 was overexpressed in transfected SGC7901 cells, accompanied by downregulation of NFκB p65 expression in the tranfectants. EMSA and MTT demonstrated that Smad4 overexpression significantly inhibited the activation of NFκB and the proliferation of SGC7901 cells (P<0.05, P<0.01).Conclusion: Smad4 overexpression can greatly inhibit the proliferation of human gastric cancer cells, probably through downregulation of NFκB pathway.
2009, 16(3):253-257.
DOI: 10.3872/j.issn.1007-385X.2009.3.009
Abstract:
Objective :To construct hepatocarcinoma specific IL1β antisense RNA expression vector and to explore its effect on the growth of implanted hepatocarcinoma H22 cells in mice and the possible mechanism. Methods: Murine IL1β antisense RNA expression vectors pafpIRES2antiIL1β1 and pafpIRES2antiIL1β2 under the regulation of minimal alphafeto protein (AFP) promoter and CMV enhancer were constructed, and further verified by PCR, restriction endonuclease analysis and DNA sequencing. H22 cells transfected with pafpIRES2antiIL1β 1 or pafpIRES2antiIL1β 2 were divided into 3 groups: H22/mock, H22/antiIL1β1 and H22/antiIL1β2 group. Expression of IL1β was detected by RTPCR. Transfected H22 cells were subcutaneously injected into mice to establish tumor implanted mouse model. Tumor volume was measured; the cytotocixity of spleen NK against H22 cells was detected by MTT. Results: Hepatocarcinoma specific IL1β antisense RNA expression vectors pafpIRES2antiIL1β1 and pafpIRES2antiIL1β2 were successfully constructed and were verified by PCR, restriction endonuclease analysis and DNA sequencing. IL1β expression in H22 cells was downregulated after transfected with IL1β antisense RNA expression vectors, especially with the pafpIRES2antiIL1β2 vector. Hepatocarcinoma cells implanted mouse model was successfully established. Tumor volume and growth of tumor in H22/antiIL1β2 mice was obviously smaller than that in H22/mock mice, and the cytotocixity of spleen NK against H22 cells in H22/antiIL1β1 and H22/antiIL1β2 mice was also greatly enhanced. Conclusion: Hepatocarcinoma specific IL1β antisense RNA expression vector pafpIRES2antiIL1β was successfully constructed. It effectively inhibits the growth of implanted hepatocarcinoma in mice probably through specifically blocking expression of IL1β and increasing cytotocixity of spleen NK.
Objective :To construct hepatocarcinoma specific IL1β antisense RNA expression vector and to explore its effect on the growth of implanted hepatocarcinoma H22 cells in mice and the possible mechanism. Methods: Murine IL1β antisense RNA expression vectors pafpIRES2antiIL1β1 and pafpIRES2antiIL1β2 under the regulation of minimal alphafeto protein (AFP) promoter and CMV enhancer were constructed, and further verified by PCR, restriction endonuclease analysis and DNA sequencing. H22 cells transfected with pafpIRES2antiIL1β 1 or pafpIRES2antiIL1β 2 were divided into 3 groups: H22/mock, H22/antiIL1β1 and H22/antiIL1β2 group. Expression of IL1β was detected by RTPCR. Transfected H22 cells were subcutaneously injected into mice to establish tumor implanted mouse model. Tumor volume was measured; the cytotocixity of spleen NK against H22 cells was detected by MTT. Results: Hepatocarcinoma specific IL1β antisense RNA expression vectors pafpIRES2antiIL1β1 and pafpIRES2antiIL1β2 were successfully constructed and were verified by PCR, restriction endonuclease analysis and DNA sequencing. IL1β expression in H22 cells was downregulated after transfected with IL1β antisense RNA expression vectors, especially with the pafpIRES2antiIL1β2 vector. Hepatocarcinoma cells implanted mouse model was successfully established. Tumor volume and growth of tumor in H22/antiIL1β2 mice was obviously smaller than that in H22/mock mice, and the cytotocixity of spleen NK against H22 cells in H22/antiIL1β1 and H22/antiIL1β2 mice was also greatly enhanced. Conclusion: Hepatocarcinoma specific IL1β antisense RNA expression vector pafpIRES2antiIL1β was successfully constructed. It effectively inhibits the growth of implanted hepatocarcinoma in mice probably through specifically blocking expression of IL1β and increasing cytotocixity of spleen NK.
2009, 16(3):258-262.
DOI: 10.3872/j.issn.1007-385X.2009.3.010
Abstract:
Objective:To construct recombinant replication defective adenoviral vectors encoding 1/100 and 1/1000 attenuated Shigalike toxinⅠ(Stx1) gene, and to study their therapeutic effects against breast cancer T47D cells in vivo.Methods: Genes encoding 1/100 or 1/1000 attenuated Stx1 were amplified by overlapping PCR and were cloned into T vectors. The inserted gene was verified by nucleotide sequencing. Replication defective adenoviral vector AdvStx1R170L containing 1/1000 attenuated Stx1 mutant gene was generated by AdMAX Adenoviral Vector System. Nude mouse models bearing human breast cancer T47D cells were established, and the tumor inhibitory effect of AdvStx1R170L was studied by intratumor injection of the recombinant adenovirus. Results: Vectors carrying 1/100 or 1/1000 attenuated Stx1 gene were successfully constructed and were verified by nucleotide sequencing. Recombinant replication defective adenoviral vector AdvStx1R170L containing 1/1000 attenuated Shigalike toxinⅠgene was constructed. In vivo study showed that AdvStx1R170L significantly inhibited the growth of implanted T47D tumor in the nude mice compared with AdvGFP and PBS group（P<0.05）. Conclusion: Recombinant replication defective adenoviral vector AdvStx1R170L encoding 1/1000 attenuated Shigalike toxin Ⅰgene has been successfully constructed, and it can effectively inhibit the growth of implanted T47D tumor in nude mice, without obvious toxicity.
Objective:To construct recombinant replication defective adenoviral vectors encoding 1/100 and 1/1000 attenuated Shigalike toxinⅠ(Stx1) gene, and to study their therapeutic effects against breast cancer T47D cells in vivo.Methods: Genes encoding 1/100 or 1/1000 attenuated Stx1 were amplified by overlapping PCR and were cloned into T vectors. The inserted gene was verified by nucleotide sequencing. Replication defective adenoviral vector AdvStx1R170L containing 1/1000 attenuated Stx1 mutant gene was generated by AdMAX Adenoviral Vector System. Nude mouse models bearing human breast cancer T47D cells were established, and the tumor inhibitory effect of AdvStx1R170L was studied by intratumor injection of the recombinant adenovirus. Results: Vectors carrying 1/100 or 1/1000 attenuated Stx1 gene were successfully constructed and were verified by nucleotide sequencing. Recombinant replication defective adenoviral vector AdvStx1R170L containing 1/1000 attenuated Shigalike toxinⅠgene was constructed. In vivo study showed that AdvStx1R170L significantly inhibited the growth of implanted T47D tumor in the nude mice compared with AdvGFP and PBS group（P<0.05）. Conclusion: Recombinant replication defective adenoviral vector AdvStx1R170L encoding 1/1000 attenuated Shigalike toxin Ⅰgene has been successfully constructed, and it can effectively inhibit the growth of implanted T47D tumor in nude mice, without obvious toxicity.
2009, 16(3):263-266.
DOI: 10.3872/j.issn.1007-385X.2009.3.011
Abstract:
objective: To study the in vivo antitumor effect of exosomes (Exo) combined with bacillus CalmetteGuérin vaccine（BCG）. Methods:Exo was isolated and purified from culture supernatant of E.G7OVA tumor cells by density gradient centrifugation. Protein components of Exo were detected by Western blotting. Exo, BCG, Exo combined with BCG (Exo+BCG) or PBS were preinjected into mice before injection of E.G7OVA cells, and the antitumor effects were observed in each group. Mouse model bearing E.G7OVA cells was established to examine the immunotherapy effects of Exo with or without BCG. Cytotoxity of spleen CTL was measured by LDH in different groups. Results: Exo derived from E.G7OVA cells contained HSP60, OVA, HSC70 and CD63 as detected by Western blotting. Tumorfree rate at 90 d was significantly higher in Exo+BCG vaccinated mice than those in Exo or BCG vaccinated mice as measured by immunoprotective assay (60% vs 20% or 0%，P<0.01). Immunotherapy assay showed that tumor inhibitory effect in Exo+BCG group was significantly higher than those in Exo or BCG groups (P<0.01). CTL results showed that CTL of Exo+BCG vaccinated mice had significantly enhanced ability to specifically kill target E.G7OVA cells compared with those of Exo and BCG groups (P<0.01). Conclusion: BCG as an immunoadjuvant can significantly enhance the antitumor effect of exosomes in vivo.
objective: To study the in vivo antitumor effect of exosomes (Exo) combined with bacillus CalmetteGuérin vaccine（BCG）. Methods:Exo was isolated and purified from culture supernatant of E.G7OVA tumor cells by density gradient centrifugation. Protein components of Exo were detected by Western blotting. Exo, BCG, Exo combined with BCG (Exo+BCG) or PBS were preinjected into mice before injection of E.G7OVA cells, and the antitumor effects were observed in each group. Mouse model bearing E.G7OVA cells was established to examine the immunotherapy effects of Exo with or without BCG. Cytotoxity of spleen CTL was measured by LDH in different groups. Results: Exo derived from E.G7OVA cells contained HSP60, OVA, HSC70 and CD63 as detected by Western blotting. Tumorfree rate at 90 d was significantly higher in Exo+BCG vaccinated mice than those in Exo or BCG vaccinated mice as measured by immunoprotective assay (60% vs 20% or 0%，P<0.01). Immunotherapy assay showed that tumor inhibitory effect in Exo+BCG group was significantly higher than those in Exo or BCG groups (P<0.01). CTL results showed that CTL of Exo+BCG vaccinated mice had significantly enhanced ability to specifically kill target E.G7OVA cells compared with those of Exo and BCG groups (P<0.01). Conclusion: BCG as an immunoadjuvant can significantly enhance the antitumor effect of exosomes in vivo.
2009, 16(3):267-271.
DOI: 10.3872/j.issn.1007-385X.2009.3.012
Abstract:
Objective:To investigate the inhibitory effect of cytokineinduced killer cells (CIK) against implanted gastric cancer cells. Methods: Gastric cancer SGC7901 cells were subcutaneously injected into the inguina of nude mice to establish gastric cancer model. The tumor bearing mice were randomly divided into CIK group and fibroblasts group, in which mice were subcutaneously injected with fluorescence dye SPDiI labeled CIK and fibroblasts HFLI cells, respectively. Distribution of CIK and HFLI cells in different tissues of gastric cancer bearing mice were observed. Meanwhile, tumor volume was measured after different treatments and tumor inhibitory rate was calculated. Tumor necrosis areas in different groups were observed. Results:SPDiI labeled CIK was mainly located in the gastric cancer tissues 10 d after injection, and was hardly detected at the injection sites, liver, spleen and lung tissues (P<0.01); SPDiI labeled fibroblasts were not found in tumors, liver, spleen and lung tissues, and were mainly located in the injection sites. Volume of implanted tumor in CIK treated mice was significantly smaller than that in the control group (P<0.05), and tumor inhibitory rate of CIK group was 29.82%. The necrosis area score of implanted tumors was significantly higher in CIK group compared with that in the control group (P<0.01). Conclusion: CIK exhibits satisfactory ability to specifically kill implanted gastric cancer.
Objective:To investigate the inhibitory effect of cytokineinduced killer cells (CIK) against implanted gastric cancer cells. Methods: Gastric cancer SGC7901 cells were subcutaneously injected into the inguina of nude mice to establish gastric cancer model. The tumor bearing mice were randomly divided into CIK group and fibroblasts group, in which mice were subcutaneously injected with fluorescence dye SPDiI labeled CIK and fibroblasts HFLI cells, respectively. Distribution of CIK and HFLI cells in different tissues of gastric cancer bearing mice were observed. Meanwhile, tumor volume was measured after different treatments and tumor inhibitory rate was calculated. Tumor necrosis areas in different groups were observed. Results:SPDiI labeled CIK was mainly located in the gastric cancer tissues 10 d after injection, and was hardly detected at the injection sites, liver, spleen and lung tissues (P<0.01); SPDiI labeled fibroblasts were not found in tumors, liver, spleen and lung tissues, and were mainly located in the injection sites. Volume of implanted tumor in CIK treated mice was significantly smaller than that in the control group (P<0.05), and tumor inhibitory rate of CIK group was 29.82%. The necrosis area score of implanted tumors was significantly higher in CIK group compared with that in the control group (P<0.01). Conclusion: CIK exhibits satisfactory ability to specifically kill implanted gastric cancer.
2009, 16(3):272-276.
DOI: 10.3872/j.issn.1007-385X.2009.3.013
Abstract:
Objective:To examine the serum proteomic spectra of human esophagial carcinoma by matrixassisted laser desorption/ionization timeofflight mass spectrometry (MALDITOF MS), so as to set up a diagnostic model of esophagial carcinoma and to investigate its clinical value. Methods: Thirtytwo esophagial carcinoma patients and 28 healthy controls were obtained from Fourth Affiliated Hospital of Hebei Medical University during May to September of 2008. Serum protein was extracted by weak cation exchange (WCX) protein chip system, and proteomic spectra was examined by MALDITOF MS. The obtained data were analyzed by ZUCIprotein chip data analyze system (ZUCIPCDAS) and an esophagial carcinoma diagnostic model was established by genetic arithmetic (GA) combined support vector machine (SVM). The above 60 samples were randomly divided into training set and blinding test set, with training set including 21 esophagial carcinoma patients and 19 healthy controls and blinding test set including 11 esophagial carcinoma patients and 9 healthy controls, so as to examine the specificity and sensitivity of this diagnostic model. Results: Serum proteomic spectra of esophagial carcinoma patients and healthy controls were obtained by MALDITOF MS, and m/z (mass to charge) peaks of 44 differential proteins were obtained after analyzed by ZUCIPCDAS software package (P<0.05). From which, 6 differential proteins (whose m/z peaks being 2 210, 2 864, 6 634, 4 068, 2 083 and 8 131, respectively) were selected to establish a diagnostic model of esophagial carcinoma. Specificity and sensitivity of this model in diagnosing esophagial carcinoma was 88.9% and 100%, respectively. Conclusion: An esophagial carcinoma diagnostic model has been established from serum proteomic spectra of esophagial carcinoma by MALDITOF MS and has high sensitivity and specificity to diagnose esophagial carcinoma.
Objective:To examine the serum proteomic spectra of human esophagial carcinoma by matrixassisted laser desorption/ionization timeofflight mass spectrometry (MALDITOF MS), so as to set up a diagnostic model of esophagial carcinoma and to investigate its clinical value. Methods: Thirtytwo esophagial carcinoma patients and 28 healthy controls were obtained from Fourth Affiliated Hospital of Hebei Medical University during May to September of 2008. Serum protein was extracted by weak cation exchange (WCX) protein chip system, and proteomic spectra was examined by MALDITOF MS. The obtained data were analyzed by ZUCIprotein chip data analyze system (ZUCIPCDAS) and an esophagial carcinoma diagnostic model was established by genetic arithmetic (GA) combined support vector machine (SVM). The above 60 samples were randomly divided into training set and blinding test set, with training set including 21 esophagial carcinoma patients and 19 healthy controls and blinding test set including 11 esophagial carcinoma patients and 9 healthy controls, so as to examine the specificity and sensitivity of this diagnostic model. Results: Serum proteomic spectra of esophagial carcinoma patients and healthy controls were obtained by MALDITOF MS, and m/z (mass to charge) peaks of 44 differential proteins were obtained after analyzed by ZUCIPCDAS software package (P<0.05). From which, 6 differential proteins (whose m/z peaks being 2 210, 2 864, 6 634, 4 068, 2 083 and 8 131, respectively) were selected to establish a diagnostic model of esophagial carcinoma. Specificity and sensitivity of this model in diagnosing esophagial carcinoma was 88.9% and 100%, respectively. Conclusion: An esophagial carcinoma diagnostic model has been established from serum proteomic spectra of esophagial carcinoma by MALDITOF MS and has high sensitivity and specificity to diagnose esophagial carcinoma.
2009, 16(3):277-281.
DOI: 10.3872/j.issn.1007-385X.2009.3.014
Abstract:
Objective:To investigate the expression of negative costimulatory molecules B7H1 and B7H3 in breast cancer and its relationship with patient's clinical parameters, prognosis and infiltration of CD3+ T lymphocytes. Methods: Fortynine breast cancer patients, who were diagnosed as having infiltrating ductal breast cancer histopathological were selected from Third Affiliated Hospital of Suzhou University from March 2003 to January 2004. B7H1 and B7H3 expression and CD3+ T lymphocytes infiltration in breast cancer tissues were detected by immunohistochemistry. Results: (1) B7H1 positive expression rate was 53.06%(26/49) in breast cancer tissues; the expression was positively correlated with the tumor size (P<0.05) and Her2/neu expression (P<0.05), and negatively with the intensity of CD3+ T infiltration (P<0.05). (2) B7H3 positive expression rate was 59.18%（29/49）in breast cancer tissues; the expression was positively correlated with patient′s pathological stage (P<0.05) and negatively with postoperative prognosis (P<0.05). (3) B7H1 expression in breast cancers was positively correlated with B7H3 expression (r=0.3316, P<0.05). Conclusion:Expression of negative costimulatory molecules B7H1 and B7H3 in breast caner is significantly correlated with the clinicopathological parameters and postoperative prognosis of patients. B7H1 and B7H3 might have a potential role in clinical diagnosis and prognostic evaluation of breast cancers.
Objective:To investigate the expression of negative costimulatory molecules B7H1 and B7H3 in breast cancer and its relationship with patient's clinical parameters, prognosis and infiltration of CD3+ T lymphocytes. Methods: Fortynine breast cancer patients, who were diagnosed as having infiltrating ductal breast cancer histopathological were selected from Third Affiliated Hospital of Suzhou University from March 2003 to January 2004. B7H1 and B7H3 expression and CD3+ T lymphocytes infiltration in breast cancer tissues were detected by immunohistochemistry. Results: (1) B7H1 positive expression rate was 53.06%(26/49) in breast cancer tissues; the expression was positively correlated with the tumor size (P<0.05) and Her2/neu expression (P<0.05), and negatively with the intensity of CD3+ T infiltration (P<0.05). (2) B7H3 positive expression rate was 59.18%（29/49）in breast cancer tissues; the expression was positively correlated with patient′s pathological stage (P<0.05) and negatively with postoperative prognosis (P<0.05). (3) B7H1 expression in breast cancers was positively correlated with B7H3 expression (r=0.3316, P<0.05). Conclusion:Expression of negative costimulatory molecules B7H1 and B7H3 in breast caner is significantly correlated with the clinicopathological parameters and postoperative prognosis of patients. B7H1 and B7H3 might have a potential role in clinical diagnosis and prognostic evaluation of breast cancers.
2009, 16(3):282-286.
DOI: 10.3872/j.issn.1007-385X.2009.3.015
Abstract:
Objective: To examine the expression of receptor tyrosine kinase（RTK）Axl and its ligand Gas6 (growth arrestspecific gene 6) protein in prostate cancer (PCa) tissues and their relation with pathological characteristics of PCa. Methods: Fortyfive PCa tissues and 32 benign prostatic hyperplasia tissues (2007 to 2008, Changhai Hospital) were obtained during surgery or kidney puncture. Expression of Axl and Gas6 protein in PCa cell lines LNCaP, PC3 and DU145 was analyzed by Western blotting. Expression of Axl and Gas6 protein in PCa tissues or benign prostatic hyperplasia tissues was examined by immunohistochemisty. Results: Axl expression was higher in highly invasive DU145 and PC3 cells than that in lowly invasive LNCaP cells. Expression of Gas6 in DU145, PC3 and LNCaP cells was similar. Expression of Axl and Gas6 protein in PCa tissues was significantly higher than that in benign prostatic hyperplasia tissues as detected by immunohistochemisty（68.9％ vs 21.9％; 53.3% vs 15.6％, all P<0.01）. Expression of Axl and Gas6 protein in PCa tissues was closely correlated with Gleason scores and metastasis of PCa (P<0.05). Conclusion: Expression of Axl and Gas6 in PCa is closely correlated with the malignancy degree and metastasis of PCa.
Objective: To examine the expression of receptor tyrosine kinase（RTK）Axl and its ligand Gas6 (growth arrestspecific gene 6) protein in prostate cancer (PCa) tissues and their relation with pathological characteristics of PCa. Methods: Fortyfive PCa tissues and 32 benign prostatic hyperplasia tissues (2007 to 2008, Changhai Hospital) were obtained during surgery or kidney puncture. Expression of Axl and Gas6 protein in PCa cell lines LNCaP, PC3 and DU145 was analyzed by Western blotting. Expression of Axl and Gas6 protein in PCa tissues or benign prostatic hyperplasia tissues was examined by immunohistochemisty. Results: Axl expression was higher in highly invasive DU145 and PC3 cells than that in lowly invasive LNCaP cells. Expression of Gas6 in DU145, PC3 and LNCaP cells was similar. Expression of Axl and Gas6 protein in PCa tissues was significantly higher than that in benign prostatic hyperplasia tissues as detected by immunohistochemisty（68.9％ vs 21.9％; 53.3% vs 15.6％, all P<0.01）. Expression of Axl and Gas6 protein in PCa tissues was closely correlated with Gleason scores and metastasis of PCa (P<0.05). Conclusion: Expression of Axl and Gas6 in PCa is closely correlated with the malignancy degree and metastasis of PCa.
2009, 16(3):287-291.
DOI: 10.3872/j.issn.1007-385X.2009.3.016
Abstract:
Objective:To study the expression of Ets1 and VEGF in breast invasive ductal carcinoma (IDC) tissues, and to analyze its correlation with clinicopathologic characteristics of IDC. Methods: Forty breast IDC tissues and their adjacent normal tissue samples were obtained from clinical diagnosed breast IDC patients after surgery. Expression of Ets1 and VEGF protein and mRNA was examined by immunohistochemistry and RTPCR. Results:（1）Expression of Ets1 and VEGF protein in breast IDC tissues was significantly higher than that in adjacent normal tissues (P<0.01). Expression of Ets1 protein in breast IDC tissues was significantly higher than that of VEGF and their expressions were positively correlated (r=0.8827, P<0.05). Expression of Ets1 and VEGF protein was correlated with the clinical stages of breast IDC and lymph node metastasis (P<0.05).（2）Expression of Ets1 and VEGF mRNA in breast IDC tissues was significantly higher than that in adjacent normal tissues (P<0.01). Expression of Ets1 mRNA in breast IDC tissues was significantly higher than the expression of VEGF and they were positively correlated (r=0.984, P<0.01). Expression of Ets1 and VEGF mRNA was correlated with the clinical stage and lymph node metastasis of breast IDC (P<005), but not with patient′s ages and tumor volumes. Conclusion: Ets1 and VEGF are highly expressed in breast IDC tissues at both mRNA and protein, and they are positively correlated with each other. Their expression is associated with the clinical stages and lymph node metastasis of breast IDC.
Objective:To study the expression of Ets1 and VEGF in breast invasive ductal carcinoma (IDC) tissues, and to analyze its correlation with clinicopathologic characteristics of IDC. Methods: Forty breast IDC tissues and their adjacent normal tissue samples were obtained from clinical diagnosed breast IDC patients after surgery. Expression of Ets1 and VEGF protein and mRNA was examined by immunohistochemistry and RTPCR. Results:（1）Expression of Ets1 and VEGF protein in breast IDC tissues was significantly higher than that in adjacent normal tissues (P<0.01). Expression of Ets1 protein in breast IDC tissues was significantly higher than that of VEGF and their expressions were positively correlated (r=0.8827, P<0.05). Expression of Ets1 and VEGF protein was correlated with the clinical stages of breast IDC and lymph node metastasis (P<0.05).（2）Expression of Ets1 and VEGF mRNA in breast IDC tissues was significantly higher than that in adjacent normal tissues (P<0.01). Expression of Ets1 mRNA in breast IDC tissues was significantly higher than the expression of VEGF and they were positively correlated (r=0.984, P<0.01). Expression of Ets1 and VEGF mRNA was correlated with the clinical stage and lymph node metastasis of breast IDC (P<005), but not with patient′s ages and tumor volumes. Conclusion: Ets1 and VEGF are highly expressed in breast IDC tissues at both mRNA and protein, and they are positively correlated with each other. Their expression is associated with the clinical stages and lymph node metastasis of breast IDC.
2009, 16(3):292-295.
DOI: 10.3872/j.issn.1007-385X.2009.3.017
Abstract:
Objective ：To examine the expression of ERCC1 (excision repair crosscomplementing 1, ERCC1) in nasopharyngeal carcinoma (NPC) tissues and its relationship with chemosensitivity to cisplatinbased chemotherapy. Methods: Eightytwo patients with advanced NPC, who were diagnosed at Nanxisan hospital of Guangxizhuang autonomous region from June 2006 to June 2008, were treated with cisplatin+5FU regimen. The expression of ERCC1 in 82 nasopharyngeal carcinoma (NPC) tissues and in 34 adjacent normal tissues was detected by immunohistochemistry assay. Results: The positive rate of ERCC1 expression in NPC adjacent normal tissues was significantly higher than that in NPC tissues (88.2% vs 63.4%, P<0.05). Expression of ERCC1 in NPC was positively correlated with patients′ age, but not with gender, T stage, N stage and M stage (P>0.05). The efficacy rate of cisplatinbased chemotherapy in ERCC1 positive NPC patients was significantly higher than that in ERCC1 negative patients (63.3% vs 36.5%, P<0.05). Conclusion: ERCC1 is lowly expressed in advanced NPC tissues; its expression is negatively correlated with the chemosensitivity to cisplatin+5FU regimen. ERCC1 can be used to predict the sensitivity of NPC patients to cisplatinbased chemotherapy.
Objective ：To examine the expression of ERCC1 (excision repair crosscomplementing 1, ERCC1) in nasopharyngeal carcinoma (NPC) tissues and its relationship with chemosensitivity to cisplatinbased chemotherapy. Methods: Eightytwo patients with advanced NPC, who were diagnosed at Nanxisan hospital of Guangxizhuang autonomous region from June 2006 to June 2008, were treated with cisplatin+5FU regimen. The expression of ERCC1 in 82 nasopharyngeal carcinoma (NPC) tissues and in 34 adjacent normal tissues was detected by immunohistochemistry assay. Results: The positive rate of ERCC1 expression in NPC adjacent normal tissues was significantly higher than that in NPC tissues (88.2% vs 63.4%, P<0.05). Expression of ERCC1 in NPC was positively correlated with patients′ age, but not with gender, T stage, N stage and M stage (P>0.05). The efficacy rate of cisplatinbased chemotherapy in ERCC1 positive NPC patients was significantly higher than that in ERCC1 negative patients (63.3% vs 36.5%, P<0.05). Conclusion: ERCC1 is lowly expressed in advanced NPC tissues; its expression is negatively correlated with the chemosensitivity to cisplatin+5FU regimen. ERCC1 can be used to predict the sensitivity of NPC patients to cisplatinbased chemotherapy.
2009, 16(3):296-299.
DOI: 10.3872/j.issn.1007-385X.2009.3.018
Abstract:
Objective:To observe the survival rate and activity of dendritic cells (DCs) to induce the activation of cytokine induced killer cell (CIK) after DCs being preserved in modified cell freezing medium (CFM). Methods:PBMCderived DCs were preserved in three different CFMs at -80 ℃ and -196 ℃, respectively. CFM I was RPMI 1640 containing 10% DMSO, 20% FCS; CFM Ⅱ was Cellbanker (ZENOAQ company, Japan); CFM Ⅲ was modified CFM containing DMSO, hydroxyethylamyle and cell stabilizer. Survival rate of DCs and antitumor activity of DCs activatied CIK (DCCIK) were measured by Trypan blue dye exclusion method and MTT after DCs being preserved in three different CFMs for 30, 60 and 90 d at -80 ℃ or -196 ℃, respectively. Results: Survival rate and activity of DCs preserved in three different CFMs were only slightly reduced as storage period was increased, and there were no significant differences among three CFMs. Conclusion:Modified CFM can substitute traditional and imported Cellbanker CFM in DCs freezing with bright future.
Objective:To observe the survival rate and activity of dendritic cells (DCs) to induce the activation of cytokine induced killer cell (CIK) after DCs being preserved in modified cell freezing medium (CFM). Methods:PBMCderived DCs were preserved in three different CFMs at -80 ℃ and -196 ℃, respectively. CFM I was RPMI 1640 containing 10% DMSO, 20% FCS; CFM Ⅱ was Cellbanker (ZENOAQ company, Japan); CFM Ⅲ was modified CFM containing DMSO, hydroxyethylamyle and cell stabilizer. Survival rate of DCs and antitumor activity of DCs activatied CIK (DCCIK) were measured by Trypan blue dye exclusion method and MTT after DCs being preserved in three different CFMs for 30, 60 and 90 d at -80 ℃ or -196 ℃, respectively. Results: Survival rate and activity of DCs preserved in three different CFMs were only slightly reduced as storage period was increased, and there were no significant differences among three CFMs. Conclusion:Modified CFM can substitute traditional and imported Cellbanker CFM in DCs freezing with bright future.
2009, 16(3):300-300.
DOI: 10.3872/j.issn.1007-385X.2009.3.019
Abstract:
Toll样受体（toll like receptors，TLRs）识别各自的配体，是机体抵抗感染性疾病和肿瘤的第一道防线。TLRs主要在固有性免疫系统细胞表面表达，一些非免疫细胞也表达TLRs;不同TLRs定位于不同的细胞。TLRs信号转导通路分为MyD88依赖性和MyD88非依赖性两种。已证实几种TLRs的激动剂具有抗肿瘤作用，肿瘤细胞表面TLRs的活化也可能对肿瘤生长产生影响。近年来TLRs在血液系统恶性肿瘤如多发性骨髓瘤、白血病和淋巴瘤方面的研究已取得一定进展，多发性骨髓瘤、白血病细胞表达多种TLRs，TLRs激动剂能诱导多发性骨髓瘤细胞发生免疫逃逸，最近白血病的免疫治疗方案中已包含TRLs激动剂，TRLs激动剂用于淋巴瘤的治疗目前正在进行临床试验。
Toll样受体（toll like receptors，TLRs）识别各自的配体，是机体抵抗感染性疾病和肿瘤的第一道防线。TLRs主要在固有性免疫系统细胞表面表达，一些非免疫细胞也表达TLRs;不同TLRs定位于不同的细胞。TLRs信号转导通路分为MyD88依赖性和MyD88非依赖性两种。已证实几种TLRs的激动剂具有抗肿瘤作用，肿瘤细胞表面TLRs的活化也可能对肿瘤生长产生影响。近年来TLRs在血液系统恶性肿瘤如多发性骨髓瘤、白血病和淋巴瘤方面的研究已取得一定进展，多发性骨髓瘤、白血病细胞表达多种TLRs，TLRs激动剂能诱导多发性骨髓瘤细胞发生免疫逃逸，最近白血病的免疫治疗方案中已包含TRLs激动剂，TRLs激动剂用于淋巴瘤的治疗目前正在进行临床试验。
2009, 16(3):301-304.
DOI: 10.3872/j.issn.1007-385X.2009.3.020
Abstract:
Toll样受体（toll like receptors，TLRs）识别各自的配体，是机体抵抗感染性疾病和肿瘤的第一道防线。TLRs主要在固有性免疫系统细胞表面表达，一些非免疫细胞也表达TLRs;不同TLRs定位于不同的细胞。TLRs信号转导通路分为MyD88依赖性和MyD88非依赖性两种。已证实几种TLRs的激动剂具有抗肿瘤作用，肿瘤细胞表面TLRs的活化也可能对肿瘤生长产生影响。近年来TLRs在血液系统恶性肿瘤如多发性骨髓瘤、白血病和淋巴瘤方面的研究已取得一定进展，多发性骨髓瘤、白血病细胞表达多种TLRs，TLRs激动剂能诱导多发性骨髓瘤细胞发生免疫逃逸，最近白血病的免疫治疗方案中已包含TRLs激动剂，TRLs激动剂用于淋巴瘤的治疗目前正在进行临床试验。
Toll样受体（toll like receptors，TLRs）识别各自的配体，是机体抵抗感染性疾病和肿瘤的第一道防线。TLRs主要在固有性免疫系统细胞表面表达，一些非免疫细胞也表达TLRs;不同TLRs定位于不同的细胞。TLRs信号转导通路分为MyD88依赖性和MyD88非依赖性两种。已证实几种TLRs的激动剂具有抗肿瘤作用，肿瘤细胞表面TLRs的活化也可能对肿瘤生长产生影响。近年来TLRs在血液系统恶性肿瘤如多发性骨髓瘤、白血病和淋巴瘤方面的研究已取得一定进展，多发性骨髓瘤、白血病细胞表达多种TLRs，TLRs激动剂能诱导多发性骨髓瘤细胞发生免疫逃逸，最近白血病的免疫治疗方案中已包含TRLs激动剂，TRLs激动剂用于淋巴瘤的治疗目前正在进行临床试验。
2009, 16(3):305-309.
DOI: 10.3872/j.issn.1007-385X.2009.3.021
Abstract:
甲状腺癌（thyroid carcinoma，TC）是内分泌系统最常见的一类恶性肿瘤，约占人类全部恶性肿瘤的1%。常见的4种病理类型中，约80%的乳头状癌（thyroid papillary carcinoma，PTC）中有BRAF突变（45%）、RET/PTC重排（20%～80%）和RAS突变（10%～20%），但很少在同一个病灶中共存。约15%的滤泡状癌（follicular thyroid carcinoma，FTC）中，常见RAS突变(40%～50%)和PAX8PPARγ点突变（35%）, 以及PI3K/AKT突变（6%～13%）；胡尔特尔细胞（Hürthle cell，HC）瘤中存在RAS突变（5%～25%）。低分化和退行性甲状腺癌中（poorly and anaplastic differentiated thyroid carcinomas; pDTC,aDTC）中，TP53点突变分别为60%～80%、15%～30%，RAS点突变分别为18%～27%、50%～60%，BRAF突变分别为15%、20%。髓样癌（medulla thyroid carcinoma，MTC）进展期常见到RET点突变。大多数基因突变会导致促分裂原活化蛋白激酶（MAPK）信号系统的激活。应用分子诊断技术检测这些基因突变，可应用于TC的病理诊断、预测TC的预后，针对突变基因的靶点抑制剂有可能成为临床治疗TC的手段之一。
甲状腺癌（thyroid carcinoma，TC）是内分泌系统最常见的一类恶性肿瘤，约占人类全部恶性肿瘤的1%。常见的4种病理类型中，约80%的乳头状癌（thyroid papillary carcinoma，PTC）中有BRAF突变（45%）、RET/PTC重排（20%～80%）和RAS突变（10%～20%），但很少在同一个病灶中共存。约15%的滤泡状癌（follicular thyroid carcinoma，FTC）中，常见RAS突变(40%～50%)和PAX8PPARγ点突变（35%）, 以及PI3K/AKT突变（6%～13%）；胡尔特尔细胞（Hürthle cell，HC）瘤中存在RAS突变（5%～25%）。低分化和退行性甲状腺癌中（poorly and anaplastic differentiated thyroid carcinomas; pDTC,aDTC）中，TP53点突变分别为60%～80%、15%～30%，RAS点突变分别为18%～27%、50%～60%，BRAF突变分别为15%、20%。髓样癌（medulla thyroid carcinoma，MTC）进展期常见到RET点突变。大多数基因突变会导致促分裂原活化蛋白激酶（MAPK）信号系统的激活。应用分子诊断技术检测这些基因突变，可应用于TC的病理诊断、预测TC的预后，针对突变基因的靶点抑制剂有可能成为临床治疗TC的手段之一。
2009, 16(3):310-314.
DOI: 10.3872/j.issn.1007-385X.2009.3.022
Abstract:
化学治疗作为肿瘤常规治疗的中坚力量，其目的是杀伤肿瘤细胞，但由于化疗药物对机体免疫系统也有一定程度的杀伤或抑制效应，因而以往普遍认为化疗与免疫治疗在功能上是拮抗的。新近研究发现，一些化疗药物可以通过增强肿瘤细胞免疫原性、促进肿瘤细胞释放内源性危险信号、以及抑制调节性T细胞（regulatory T cell，Treg）等机制增强肿瘤的免疫原性，介导抗肿瘤免疫，对肿瘤的整体化疗效果有其独特的贡献。依据化疗后肿瘤细胞免疫原性的不同变化，设计相应的治疗方案，将化疗和免疫治疗有机结合，将最终提高肿瘤的治疗效果。
化学治疗作为肿瘤常规治疗的中坚力量，其目的是杀伤肿瘤细胞，但由于化疗药物对机体免疫系统也有一定程度的杀伤或抑制效应，因而以往普遍认为化疗与免疫治疗在功能上是拮抗的。新近研究发现，一些化疗药物可以通过增强肿瘤细胞免疫原性、促进肿瘤细胞释放内源性危险信号、以及抑制调节性T细胞（regulatory T cell，Treg）等机制增强肿瘤的免疫原性，介导抗肿瘤免疫，对肿瘤的整体化疗效果有其独特的贡献。依据化疗后肿瘤细胞免疫原性的不同变化，设计相应的治疗方案，将化疗和免疫治疗有机结合，将最终提高肿瘤的治疗效果。
2009, 16(3):315-318.
DOI: 10.3872/j.issn.1007-385X.2009.3.023
Abstract:
细胞周期蛋白依赖性激酶抑制剂相关基因的突变和表达失调可直接或间接影响细胞的周期、增殖以及凋亡等功能，与肿瘤的发生发展密切相关。p16能够抑制CDK4／CDK6介导的Rb蛋白产物的磷酸化，其CpG岛异常甲基化参与宫颈癌的发生发展；p21作为p53的下游调节因子，与肿瘤血管的生成、淋巴转移及预后相关；p27作用机制复杂，与乳腺癌预后关系密切，并可恢复耐药乳腺癌细胞对他莫昔芬的敏感性；p57在细胞周期G1到S期的转变中至关重要，其表达程度与部分肿瘤的恶性程度相关，它还参与了肿瘤细胞的凋亡过程。了解这些基因的结构和作用机制，探究其在肿瘤发生、发展中所产生的作用，进而寻找有效治疗靶点，已成为肿瘤分子生物学关注的热点。
细胞周期蛋白依赖性激酶抑制剂相关基因的突变和表达失调可直接或间接影响细胞的周期、增殖以及凋亡等功能，与肿瘤的发生发展密切相关。p16能够抑制CDK4／CDK6介导的Rb蛋白产物的磷酸化，其CpG岛异常甲基化参与宫颈癌的发生发展；p21作为p53的下游调节因子，与肿瘤血管的生成、淋巴转移及预后相关；p27作用机制复杂，与乳腺癌预后关系密切，并可恢复耐药乳腺癌细胞对他莫昔芬的敏感性；p57在细胞周期G1到S期的转变中至关重要，其表达程度与部分肿瘤的恶性程度相关，它还参与了肿瘤细胞的凋亡过程。了解这些基因的结构和作用机制，探究其在肿瘤发生、发展中所产生的作用，进而寻找有效治疗靶点，已成为肿瘤分子生物学关注的热点。
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.