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Volume 19,Issue 3,2012 Table of Contents
2012, 19(3):229-238.
DOI: 10.3872/j.issn.1007-385X.2012.3.001
Abstract:
The immune system can eliminate malignant cells or prevent tumor growth by recognizing and destroying malignant cells. Under the strong selective pressure from the host immune system, malignant cells can rapidly acquire new phenotypes through high somatic mutations to evade immune surveillance. The selected tumor cell variants make use of all kinds of immunosuppressive mechanisms to establish an immunosuppressive microenvironment to resist and suppress anti-tumor immune response, allowing the tumors expend and become clinically detectable. The current tumor therapies mainly focus on the direct inhibition of tumor cell proliferation, as well as the killing and elimination of tumor cells. However, the growing evidence suggests that the conventional tumor therapies can elicit specific cellular responses that render tumor-cell death immunogenic, which may activate the innate immune signaling pathways and then induce the intrinsic anti-tumor immune responses. This potential mechanism plays a key role in the therapeutic effects of the conventional anticancer treatments, especially in preventing the recurrence of residual tumor cells. Here, we intend to demonstrate the molecular and cellular mechanisms of the anti-tumor immune responses and the immunosuppressive tumor microenvironment in the process of tumor development and the conventional anticancer treatments, especially focusing on their effects on the therapeutic effects of the conventional anticancer treatments. We also attempt to explore the rational therapeutic strategies targeting the components of tumor immune microenvironment, which may shed light on the improvement of the current conventional anticancer therapies and the development of new anticancer treatments.
The immune system can eliminate malignant cells or prevent tumor growth by recognizing and destroying malignant cells. Under the strong selective pressure from the host immune system, malignant cells can rapidly acquire new phenotypes through high somatic mutations to evade immune surveillance. The selected tumor cell variants make use of all kinds of immunosuppressive mechanisms to establish an immunosuppressive microenvironment to resist and suppress anti-tumor immune response, allowing the tumors expend and become clinically detectable. The current tumor therapies mainly focus on the direct inhibition of tumor cell proliferation, as well as the killing and elimination of tumor cells. However, the growing evidence suggests that the conventional tumor therapies can elicit specific cellular responses that render tumor-cell death immunogenic, which may activate the innate immune signaling pathways and then induce the intrinsic anti-tumor immune responses. This potential mechanism plays a key role in the therapeutic effects of the conventional anticancer treatments, especially in preventing the recurrence of residual tumor cells. Here, we intend to demonstrate the molecular and cellular mechanisms of the anti-tumor immune responses and the immunosuppressive tumor microenvironment in the process of tumor development and the conventional anticancer treatments, especially focusing on their effects on the therapeutic effects of the conventional anticancer treatments. We also attempt to explore the rational therapeutic strategies targeting the components of tumor immune microenvironment, which may shed light on the improvement of the current conventional anticancer therapies and the development of new anticancer treatments.
2012, 19(3):239-246.
DOI: 10.3872/j.issn.1007-385X.2012.3.002
Abstract:
Objective: To construct the Fc fusion protein EVP1 with high affinity to bind epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), and human epidermal growth factor receptor 2 (HER2), and to analyze its multi-target binding affinity for further study of antitumor therapy. Methods: The coding sequences of Herin and FLT-1D2 (the second Ig-like domain of FLT-1gene) were amplified by PCR individually, and joined to synthesize human IgG1 Fc fragment with point mutations via overlap PCR. Then, the fused open reading frame of Herin-Flt-1D2-Fc (named as EVP1) was inserted into pDC659 plasmid, an adenoviral shuttle vector. The plasmid of pDC659-EVP1 and pPE3-F35 were co-transfected into 293 cells to package Ad5/35-EVP1, a replication-incompetent adenovirus carrying the EVP1 coding gene. After identification, amplification, purification and titration determination, the recombinant virus Ad5/35-EVP1 was used to transfect 293 cells at a MOI=11. Four days post transfection, culture supernatant was collected to enrich and purify the fusion protein EVP1 through ammonium sulfate salting out method and protein G affinity chromatography. Subsequently, Western blotting and ELISA were performed to detect the expression of EVP1. Finally, the binding affinities of EVP1 to EGFR, HER2 and VEGF were both qualitatively identified via indirect immunofluorescent assay and quantitatively determined on the Octet Red 96 Biomolecular Interaction Analysis System. Results: The recombinant adenovirus Ad5/35-EVP1 was constructed successfully, and its titer was 5.4×109 PFU/ml. With the expression system of Ad5/35-EVP1 and 293 cells, the fusion protein EVP1 could be effectively produced with the expression level being (1 613.94?24.65) ng/ml when MOI=11. The obtained EVP1 could bind EGFR on A431 cells and HER2 on SK-OV-3 cells with high affinity, and the affinity constants Kd of EGFR, HER2 and VEGF were 4.55,18.70, and 0.63 nmol/L, respectively. Conclusion: The fusion protein EVP1 was constructed successfully, and EVP1 has high binding-affinity to VEGF and EGFR family members, showing a good clinical potent in targeted cancer therapy.
Objective: To construct the Fc fusion protein EVP1 with high affinity to bind epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), and human epidermal growth factor receptor 2 (HER2), and to analyze its multi-target binding affinity for further study of antitumor therapy. Methods: The coding sequences of Herin and FLT-1D2 (the second Ig-like domain of FLT-1gene) were amplified by PCR individually, and joined to synthesize human IgG1 Fc fragment with point mutations via overlap PCR. Then, the fused open reading frame of Herin-Flt-1D2-Fc (named as EVP1) was inserted into pDC659 plasmid, an adenoviral shuttle vector. The plasmid of pDC659-EVP1 and pPE3-F35 were co-transfected into 293 cells to package Ad5/35-EVP1, a replication-incompetent adenovirus carrying the EVP1 coding gene. After identification, amplification, purification and titration determination, the recombinant virus Ad5/35-EVP1 was used to transfect 293 cells at a MOI=11. Four days post transfection, culture supernatant was collected to enrich and purify the fusion protein EVP1 through ammonium sulfate salting out method and protein G affinity chromatography. Subsequently, Western blotting and ELISA were performed to detect the expression of EVP1. Finally, the binding affinities of EVP1 to EGFR, HER2 and VEGF were both qualitatively identified via indirect immunofluorescent assay and quantitatively determined on the Octet Red 96 Biomolecular Interaction Analysis System. Results: The recombinant adenovirus Ad5/35-EVP1 was constructed successfully, and its titer was 5.4×109 PFU/ml. With the expression system of Ad5/35-EVP1 and 293 cells, the fusion protein EVP1 could be effectively produced with the expression level being (1 613.94?24.65) ng/ml when MOI=11. The obtained EVP1 could bind EGFR on A431 cells and HER2 on SK-OV-3 cells with high affinity, and the affinity constants Kd of EGFR, HER2 and VEGF were 4.55,18.70, and 0.63 nmol/L, respectively. Conclusion: The fusion protein EVP1 was constructed successfully, and EVP1 has high binding-affinity to VEGF and EGFR family members, showing a good clinical potent in targeted cancer therapy.
XIE Ning
,
ZHOU Jian
,
SONG Yong-ping
,
GAO Quan-li
,
ZHANG Yan-li
,
FU Yue-wen
,
FANG Bai-jun
,
WEI Xu-dong
2012, 19(3):247-251.
DOI: 10.3872/j.issn.1007-385X.2012.3.003
Abstract:
Objective: To observe the change of cytotoxicity of NK cells on dexamethasone-resistant B-cell lymphoma cells, and study its mechanism. Methods: 20 μg/ml dexamethasone (DXM) was added to induce DXM-resistance B lymphoma SU-DHL-4 cells (SU cells). The multidrug resistance of SU cells was named SU-DXM cells. NK cells, sorted by flow cytometry was chosen as an effector, and the cytotoxicity of NK cells on SU and SU/DXM cells was analyzed through flow cytometry at E∶T was 20∶1. Real-time PCR was used to detect the gene expression of MHC class Ⅰ chain-related molecules A/B (MICA/B), UL16 binding protein (ULBP)1, ULBP2 and ULBP3 in both SU and SU/DXM cells. Results: After DXM treatment, the SU/DXM cells were established as a multi-drug-resistant cell line. Compared to SU cells, the cytotoyxicity sensitivity of NK cells on SU/DXM cells was obviously down-regulated (\[3.57±4.22\]% vs \[1138±3.51\]%, P<0.05), and the expressions of NKG2D ligand genes MICA, MICB and ULBP2 were statistically lower in SU/DXM cells (MICA: 0.017±0.006, MICB: 0.682±0.063, and ULBP: 20.773±0.066) than those in SU cells (MICA: 1.014±0.121, MICB: 1.009±0.092, and ULBP2: 0.993±0.108, P<0.05, P<0.01). Conclusion: Devamethasone has the ability to induce SU cells to multi-drug-resistant cells, SU/DXM cells, which resist the cytotoxicity mediated by NK cells, and the low gene expression of NKG2D ligands could be one important cause.
Objective: To observe the change of cytotoxicity of NK cells on dexamethasone-resistant B-cell lymphoma cells, and study its mechanism. Methods: 20 μg/ml dexamethasone (DXM) was added to induce DXM-resistance B lymphoma SU-DHL-4 cells (SU cells). The multidrug resistance of SU cells was named SU-DXM cells. NK cells, sorted by flow cytometry was chosen as an effector, and the cytotoxicity of NK cells on SU and SU/DXM cells was analyzed through flow cytometry at E∶T was 20∶1. Real-time PCR was used to detect the gene expression of MHC class Ⅰ chain-related molecules A/B (MICA/B), UL16 binding protein (ULBP)1, ULBP2 and ULBP3 in both SU and SU/DXM cells. Results: After DXM treatment, the SU/DXM cells were established as a multi-drug-resistant cell line. Compared to SU cells, the cytotoyxicity sensitivity of NK cells on SU/DXM cells was obviously down-regulated (\[3.57±4.22\]% vs \[1138±3.51\]%, P<0.05), and the expressions of NKG2D ligand genes MICA, MICB and ULBP2 were statistically lower in SU/DXM cells (MICA: 0.017±0.006, MICB: 0.682±0.063, and ULBP: 20.773±0.066) than those in SU cells (MICA: 1.014±0.121, MICB: 1.009±0.092, and ULBP2: 0.993±0.108, P<0.05, P<0.01). Conclusion: Devamethasone has the ability to induce SU cells to multi-drug-resistant cells, SU/DXM cells, which resist the cytotoxicity mediated by NK cells, and the low gene expression of NKG2D ligands could be one important cause.
2012, 19(3):252-256.
DOI: 10.3872/j.issn.1007-385X.2012.3.004
Abstract:
Objective: To investigate the inhibitiory effect of siRNA targeting vascular endothelial growth factor (VEGF) gene on proliferation of breast cancer MCF-7 cells. Methods: Small interfering RNA (siRNA) targeting VEGF, including 2 siRNAs (siRNA21/21, siRNA23/23) and 2 asymmetric siRNAs (aiRNA) (aiRNA19/21, aiRNA21/23) were designed and synthesized. The siRNA was transfected into MCF-7 cells, and cell proliferation was detected by MTT method, and the apoptosis was analyzed by flow cytometry. VEGF protein and mRNA expression levels of MCF-7 cells were detected by ELISA and RT-PCR. Results: Compared with the control group, the 4 siRNAs effectively inhibited VEGF mRNA expression (\[714±5.01\]%, \[40.0±3.11\]%, \[37.2±2.79\]%, \[11.1±0.99\]% vs \[2.4±0.11\]%, P<005), and the proliferation of MCF-7 cells (\[44.7±5.38\]%, \[38.5±5.67\]%, \[33.6±2.18\]%, \[33.1±318\]%, \[2.2±0.28\]%, P<0.05), with aiRNA21/23 of the best RNA interference effect. Compared with the control group, the apoptotic rate was significantly increased (\[49.9±4.02\]% vs \[4.7±0.91\]%, P<0.05). Conclusion:siRNA targeting VEGF can inhibit proliferation and induce apoptosis of MCF-7 cells, with asymmetric siRNA showing best effects.
Objective: To investigate the inhibitiory effect of siRNA targeting vascular endothelial growth factor (VEGF) gene on proliferation of breast cancer MCF-7 cells. Methods: Small interfering RNA (siRNA) targeting VEGF, including 2 siRNAs (siRNA21/21, siRNA23/23) and 2 asymmetric siRNAs (aiRNA) (aiRNA19/21, aiRNA21/23) were designed and synthesized. The siRNA was transfected into MCF-7 cells, and cell proliferation was detected by MTT method, and the apoptosis was analyzed by flow cytometry. VEGF protein and mRNA expression levels of MCF-7 cells were detected by ELISA and RT-PCR. Results: Compared with the control group, the 4 siRNAs effectively inhibited VEGF mRNA expression (\[714±5.01\]%, \[40.0±3.11\]%, \[37.2±2.79\]%, \[11.1±0.99\]% vs \[2.4±0.11\]%, P<005), and the proliferation of MCF-7 cells (\[44.7±5.38\]%, \[38.5±5.67\]%, \[33.6±2.18\]%, \[33.1±318\]%, \[2.2±0.28\]%, P<0.05), with aiRNA21/23 of the best RNA interference effect. Compared with the control group, the apoptotic rate was significantly increased (\[49.9±4.02\]% vs \[4.7±0.91\]%, P<0.05). Conclusion:siRNA targeting VEGF can inhibit proliferation and induce apoptosis of MCF-7 cells, with asymmetric siRNA showing best effects.
DUAN Li-fang
,
ZHANG Lian-sheng
,
YUE Ling-ling
,
CHAI Ye
,
ZENG Peng-yun
,
WU Chong-yang
,
LI Li-juan
2012, 19(3):257-260.
DOI: 10.3872/j.issn.1007-385X.2012.3.005
Abstract:
Objective: To investigate the expression of Toll-like receptor 7 (TLR7) mRNA and TLR9 mRNA in the peripheral blood plasmacytoid dendritic cells (pDCs) of patients with chronic myeloid leukemia (CML), and the interferon-α (IFN-α) secretion by pDCs. Methods: pDCs were sorted by immune magnetic beads of the patients with CML (n=30, including 15 CML untreated and 15 CML remission) from the Department of Hematology of Second Affiliated Hospital of Lanzhou University from November 2010 to July 2011 and healthy controls (n=15) from Physical Examination Center. The mRNA expressions of TLR7 and TLR9 in pDCs were measured with real time-PCR; pDCs were stimulated by CpG ODN 2216 for 24 h, and IFN-α in the supernatant was measured using ELISA. Results: The mRNA expression of TLR7 was significantly reduced in the untreated CML group (0.34±0.11) than that in the remission CML group (0.93±021, P<005) and the healthy control group (P<0.05); The mRNA expression of TLR9 was significantly reduced in the untreated CML group (0.44±0.15) than that in the remission group (0.94±0.18, P<0.05) and the healthy control group (P<0.05); IFN-α production of pDCs was significantly reduced in the untreated CML group than that in the remission group and the healthy control group (\[408.61±77.11\] vs \[611.39±84.86\], \[651.67±93.39\] ng/L, P<0.05). Conclusion:The mRNA expressions of TLR7 and TLR9are significantly reduced in CML patients, which may be the main reason of function defects of pDCs, indicating that TLR7 and TLR9 may be involved in the pathogenesis of CML.
Objective: To investigate the expression of Toll-like receptor 7 (TLR7) mRNA and TLR9 mRNA in the peripheral blood plasmacytoid dendritic cells (pDCs) of patients with chronic myeloid leukemia (CML), and the interferon-α (IFN-α) secretion by pDCs. Methods: pDCs were sorted by immune magnetic beads of the patients with CML (n=30, including 15 CML untreated and 15 CML remission) from the Department of Hematology of Second Affiliated Hospital of Lanzhou University from November 2010 to July 2011 and healthy controls (n=15) from Physical Examination Center. The mRNA expressions of TLR7 and TLR9 in pDCs were measured with real time-PCR; pDCs were stimulated by CpG ODN 2216 for 24 h, and IFN-α in the supernatant was measured using ELISA. Results: The mRNA expression of TLR7 was significantly reduced in the untreated CML group (0.34±0.11) than that in the remission CML group (0.93±021, P<005) and the healthy control group (P<0.05); The mRNA expression of TLR9 was significantly reduced in the untreated CML group (0.44±0.15) than that in the remission group (0.94±0.18, P<0.05) and the healthy control group (P<0.05); IFN-α production of pDCs was significantly reduced in the untreated CML group than that in the remission group and the healthy control group (\[408.61±77.11\] vs \[611.39±84.86\], \[651.67±93.39\] ng/L, P<0.05). Conclusion:The mRNA expressions of TLR7 and TLR9are significantly reduced in CML patients, which may be the main reason of function defects of pDCs, indicating that TLR7 and TLR9 may be involved in the pathogenesis of CML.
2012, 19(3):261-266.
DOI: 10.3872/j.issn.1007-385X.2012.3.006
Abstract:
Objective: To observe the effects of antiMRP3(scFv)-sTRAIL on apoptosis in glioblastoma multiforme (GBM) U251 cells and to study its mechanism. Methods: Different concentration of antiMRP3(scFv)-sTRAIL (3.9063 nmol/L-250 nmol/L) was added into U251 cells, and then the viability was examined by MTT assay. IC50 dose of antiMRP3(scFv)-sTRAIL was added into U251 cells, in the presence or absence of parental antiMRP3(scFv) and the TRAIL-neutralizing mAb 2E5. The apoptotic percentage of U251 cells was examined by flow cytometry, and Western blotting was used to detect caspase-3 activation and DNA fragmentation factor (DFF) degradation in U251 cells. Results: When the concentration of antiMRP3(scFv)-sTRAIL increased (3.9063 nmol/L-250 nmol/L), the viability of U251 cells became lower synchronously (\[84.84±0.2\]%-\[19.93±1.8\]%); The apoptotic rates of U251 cells induced by IC50 dose (62.5 nmol/L) of antiMRP3(scFv)-sTRAIL or antiMRP3(scFv)-sTRAIL with parental antiMRP3(scFv) and mAb 2E5 were (58.0±1.3)%, (14.9±1.7)% and (17.4±3.0)%, respectively; antiMRP3(scFv) and the mAb 2E5 significantly inhibited the activation of caspase-3 and the degradation of DFF in U251 cells induced by antiMRP3(scFv)-sTRAIL. Conclusion: Targeted binding of antiMRP3(scFv)-sTRAIL to GBM increases the apoptosis induction activity of sTRAIL, which lays a foundation for further study on tumor-targeting drugs.
Objective: To observe the effects of antiMRP3(scFv)-sTRAIL on apoptosis in glioblastoma multiforme (GBM) U251 cells and to study its mechanism. Methods: Different concentration of antiMRP3(scFv)-sTRAIL (3.9063 nmol/L-250 nmol/L) was added into U251 cells, and then the viability was examined by MTT assay. IC50 dose of antiMRP3(scFv)-sTRAIL was added into U251 cells, in the presence or absence of parental antiMRP3(scFv) and the TRAIL-neutralizing mAb 2E5. The apoptotic percentage of U251 cells was examined by flow cytometry, and Western blotting was used to detect caspase-3 activation and DNA fragmentation factor (DFF) degradation in U251 cells. Results: When the concentration of antiMRP3(scFv)-sTRAIL increased (3.9063 nmol/L-250 nmol/L), the viability of U251 cells became lower synchronously (\[84.84±0.2\]%-\[19.93±1.8\]%); The apoptotic rates of U251 cells induced by IC50 dose (62.5 nmol/L) of antiMRP3(scFv)-sTRAIL or antiMRP3(scFv)-sTRAIL with parental antiMRP3(scFv) and mAb 2E5 were (58.0±1.3)%, (14.9±1.7)% and (17.4±3.0)%, respectively; antiMRP3(scFv) and the mAb 2E5 significantly inhibited the activation of caspase-3 and the degradation of DFF in U251 cells induced by antiMRP3(scFv)-sTRAIL. Conclusion: Targeted binding of antiMRP3(scFv)-sTRAIL to GBM increases the apoptosis induction activity of sTRAIL, which lays a foundation for further study on tumor-targeting drugs.
7 Adenovirus mediated overexpression of miR-99a inhibits the growth of hepato-cellular carcinoma cells
ZHANG Jing-lei
,
JIN Hua-jun
,
LIU Hui
,
L Sai-qun
,
YANG Yuan
,
FU Si-yuan
,
QIAN Qi-jun
,
ZHOU Wei-ping
2012, 19(3):267-271.
DOI: 10.3872/j.issn.1007-385X.2012.3.007
Abstract:
Objective: To study the inhibitory effect of adenovirus-mediated-miR-99a overexpression on proliferation of hepatocellular carcinoma cells (HCCs) and find a new kind of adenovirus mediated miRNA treatment for cancer.Methods: The expression of miR-99a was detected by quantitative real-time polymerase chain reaction (qRT-PCR) in human liver cell line L-02 and HCC cell lines HepG2, SMMC-7721, Hep3B and Huh7. The type 5 adenovirus vector containing miR-99 (Ad5-miR-99a) was constructed. Huh7 cells were infected with empty adenovirus vector (Ad-blank) or Ad5-miR-99a, and MTT and colony formation assay were used to examine the inhibitory effect of miR-99a on the growth of Huh7 cells. Results: miRNA-99a was markedly decreased in the Huh7 cell line compared with that in L02 cell line (P<0.01). Ad5-miR-99a adenovirus vector was successfully constructed, and miR-99a was stably high expressed in Huh7 cells after infection with Ad5-miR-99a. MTT and colony formation assay showed that Ad5-miR-99a could inhibit the growth and colony formation of Huh7 cells compared with Ad-blank (P<0.01). Conclusion: Adenovirus Ad5-miR-99a is successfully constructed, which can effectively suppress the growth of HCC. Ad5-miR-99a might be a prospective therapeutic approach for HCC in the near future.
Objective: To study the inhibitory effect of adenovirus-mediated-miR-99a overexpression on proliferation of hepatocellular carcinoma cells (HCCs) and find a new kind of adenovirus mediated miRNA treatment for cancer.Methods: The expression of miR-99a was detected by quantitative real-time polymerase chain reaction (qRT-PCR) in human liver cell line L-02 and HCC cell lines HepG2, SMMC-7721, Hep3B and Huh7. The type 5 adenovirus vector containing miR-99 (Ad5-miR-99a) was constructed. Huh7 cells were infected with empty adenovirus vector (Ad-blank) or Ad5-miR-99a, and MTT and colony formation assay were used to examine the inhibitory effect of miR-99a on the growth of Huh7 cells. Results: miRNA-99a was markedly decreased in the Huh7 cell line compared with that in L02 cell line (P<0.01). Ad5-miR-99a adenovirus vector was successfully constructed, and miR-99a was stably high expressed in Huh7 cells after infection with Ad5-miR-99a. MTT and colony formation assay showed that Ad5-miR-99a could inhibit the growth and colony formation of Huh7 cells compared with Ad-blank (P<0.01). Conclusion: Adenovirus Ad5-miR-99a is successfully constructed, which can effectively suppress the growth of HCC. Ad5-miR-99a might be a prospective therapeutic approach for HCC in the near future.
YANG Xin-wei
,
ZHANG Lian-sheng
,
CHAI Ye
,
ZENG Peng-yun
,
WU Chong-yang
,
YUE Ling-ling
,
LI Li-juan
,
HAO Zheng-dong
,
LI Liang-Liang
2012, 19(3):272-275.
DOI: 10.3872/j.issn.1007-385X.2012.3.008
Abstract:
Objective: To study the effect of recombinant soluble MHC class Ⅰ chain-related gene A (sMICA) protein on the expression of NKG2D (natural killer group 2 member D), cytotoxicity and secretion of IFN-γ of NK cells. Methods:NK cells were separated from normal adults by magnetic cell sorting (MACS) and then divided into 4 groups: recombinant sMICA groups (200, 500, 800 μg/L groups) and a control group. After 24 h of the interaction, the NKG2D was detected by the flow cytometry; the cytotoxicity of NK cells on K562 leukemia cells was detected by LDH release assay; and the concentration of IFN-γ in supernatant was determined by ELISA. Results:Compared with the control group, NK cell surface expressions of NKG2D in 200, 500, 800 μg/L sMICA groups were down-regulated (\[68.79±6.87\]%, \[55.75±9.31\]%, \[45.14±5.70\]% vs \[92.75±6.91\]%, P<0.05 or P<0.01); sMICA inhibited thecytotoxicity of NK cells on K562 leukemia cells (\[18.67±2.3\]%, \[15.01±2.25\]%, \[7.33±2.52\]% vs \[36.33±2.51\]%, P<0.05 or P<0.01), and decreased the secretion of IFN-γ of NK cells (\[164.48±22.48\], \[112.71±10.89\], \[70.23±9.64\] pg/ml vs \[313.72±16.06\] pg/ml, P<0.05 or P<001). Conclusion: The sMICA sheddingfrom the acute leukemia cell urface can reduce the NKG2D expression and IFN-γ secretion of NK cells, thereby inhibit the ytotoxic activity of NK cells against leukemia cells.
Objective: To study the effect of recombinant soluble MHC class Ⅰ chain-related gene A (sMICA) protein on the expression of NKG2D (natural killer group 2 member D), cytotoxicity and secretion of IFN-γ of NK cells. Methods:NK cells were separated from normal adults by magnetic cell sorting (MACS) and then divided into 4 groups: recombinant sMICA groups (200, 500, 800 μg/L groups) and a control group. After 24 h of the interaction, the NKG2D was detected by the flow cytometry; the cytotoxicity of NK cells on K562 leukemia cells was detected by LDH release assay; and the concentration of IFN-γ in supernatant was determined by ELISA. Results:Compared with the control group, NK cell surface expressions of NKG2D in 200, 500, 800 μg/L sMICA groups were down-regulated (\[68.79±6.87\]%, \[55.75±9.31\]%, \[45.14±5.70\]% vs \[92.75±6.91\]%, P<0.05 or P<0.01); sMICA inhibited thecytotoxicity of NK cells on K562 leukemia cells (\[18.67±2.3\]%, \[15.01±2.25\]%, \[7.33±2.52\]% vs \[36.33±2.51\]%, P<0.05 or P<0.01), and decreased the secretion of IFN-γ of NK cells (\[164.48±22.48\], \[112.71±10.89\], \[70.23±9.64\] pg/ml vs \[313.72±16.06\] pg/ml, P<0.05 or P<001). Conclusion: The sMICA sheddingfrom the acute leukemia cell urface can reduce the NKG2D expression and IFN-γ secretion of NK cells, thereby inhibit the ytotoxic activity of NK cells against leukemia cells.
2012, 19(3):276-279.
DOI: 10.3872/j.issn.1007-385X.2012.3.009
Abstract:
Objective: To investigate the inhibitory effect of chimeric monoclonal antibody CH12 on the growth of head and neck squamous cell carcinoma (HNSCC) xenografts in vivo, and to provide basis for further study on the effect CH12 antibody on tumor therapy. Methods: The protein expression levels of the epidermal growth factor receptor (EGFR) in A253, CAL27, Detroit 562, FaDu and RPMI 2650 HNSCC cell lines were analyzed by Western blotting. The binding capacity of CH12 antibody with these five cell lines was detected by flow cytometry. CAL27 and A253 cells were subcutaneously inoculated into nude mice to establish HNSCC xenograft models. The mice with xenografted tumor were randomized into two groups: a ehicle control group (PBS) and a treatment group (CH12). Tumor volume was recorded at a regular time interval, and then the tumor growth curve was drawn. Results: EGFR was expressed in CAL27, A253,FaDu and Detroit 562 cell lines, with the highest expression in CAL27 cells and the second highest expression in A253 cells.The binding affinity of CH12 with five cell lines was followed by CAL27, FaDu, A253, Detroit 562 and RPMI 265. CH12 significantly inhibited the tumor growth in both CAL27 and A253 xenografts compared with that in the control group, and the tumor growth inhibition ratios were 56.8% (P=0.022) and 59.7% (P=0.015) respectively. Conclusion: CH12 antibody can effectively inhibit the tumor growth of EGFR high-expressed HNSCC xenografts in vivo.
Objective: To investigate the inhibitory effect of chimeric monoclonal antibody CH12 on the growth of head and neck squamous cell carcinoma (HNSCC) xenografts in vivo, and to provide basis for further study on the effect CH12 antibody on tumor therapy. Methods: The protein expression levels of the epidermal growth factor receptor (EGFR) in A253, CAL27, Detroit 562, FaDu and RPMI 2650 HNSCC cell lines were analyzed by Western blotting. The binding capacity of CH12 antibody with these five cell lines was detected by flow cytometry. CAL27 and A253 cells were subcutaneously inoculated into nude mice to establish HNSCC xenograft models. The mice with xenografted tumor were randomized into two groups: a ehicle control group (PBS) and a treatment group (CH12). Tumor volume was recorded at a regular time interval, and then the tumor growth curve was drawn. Results: EGFR was expressed in CAL27, A253,FaDu and Detroit 562 cell lines, with the highest expression in CAL27 cells and the second highest expression in A253 cells.The binding affinity of CH12 with five cell lines was followed by CAL27, FaDu, A253, Detroit 562 and RPMI 265. CH12 significantly inhibited the tumor growth in both CAL27 and A253 xenografts compared with that in the control group, and the tumor growth inhibition ratios were 56.8% (P=0.022) and 59.7% (P=0.015) respectively. Conclusion: CH12 antibody can effectively inhibit the tumor growth of EGFR high-expressed HNSCC xenografts in vivo.
2012, 19(3):280-283.
DOI: 10.3872/j.issn.1007-385X.2012.3.010
Abstract:
Objective: To study the effect of curcumin on proliferation and expression ofsurvivin, caspase-3 and p-Akt in colon cancer SW480 cells.Methods:SW480 cells were treated with different concentrations of curcumin (5-40 μmol/L) for 24, 48 and 72 h. The inhibitory effect of curcumin on SW480 cells was detected by MTT. The expressions of survivin, caspase-3 and p-Akt protein were analyzed by Western blotting. Results: Curcumin decreased the proliferation of SW480 cells in a dose- and time-dependent manner (P<0.01), with an inhibitory rate of (75.86±3.93)% after 40 μmol/L curcumin treatment for 72 h. Curcumin decreased the expressions of p-Akt and survivin, and increased the expression of caspase-3 protein in a dose- and time-dependent manner (P<0.01). Treated with curcumin (40 μmol/L) for 48 h, the expressions of p-Akt and survivin protein were decreased significantly (\[0.204±0.025\] vs \[0.367±0.035\], P<0.01; \[0.208±0.014\] vs \[0.385±0.034\], P<0.01), and the expression of caspase-3 protein was increased significantly (\[0.371±0.028\] vs \[0.127±0.023\], P<0.01). Conclusion: Cureumin inhibits the proliferation of SW480 cells, which may be related to a decrease in phosphorylation of Akt, down- regulation of survivin protein expression and up-regulation of caspase-3 protein expression.
Objective: To study the effect of curcumin on proliferation and expression ofsurvivin, caspase-3 and p-Akt in colon cancer SW480 cells.Methods:SW480 cells were treated with different concentrations of curcumin (5-40 μmol/L) for 24, 48 and 72 h. The inhibitory effect of curcumin on SW480 cells was detected by MTT. The expressions of survivin, caspase-3 and p-Akt protein were analyzed by Western blotting. Results: Curcumin decreased the proliferation of SW480 cells in a dose- and time-dependent manner (P<0.01), with an inhibitory rate of (75.86±3.93)% after 40 μmol/L curcumin treatment for 72 h. Curcumin decreased the expressions of p-Akt and survivin, and increased the expression of caspase-3 protein in a dose- and time-dependent manner (P<0.01). Treated with curcumin (40 μmol/L) for 48 h, the expressions of p-Akt and survivin protein were decreased significantly (\[0.204±0.025\] vs \[0.367±0.035\], P<0.01; \[0.208±0.014\] vs \[0.385±0.034\], P<0.01), and the expression of caspase-3 protein was increased significantly (\[0.371±0.028\] vs \[0.127±0.023\], P<0.01). Conclusion: Cureumin inhibits the proliferation of SW480 cells, which may be related to a decrease in phosphorylation of Akt, down- regulation of survivin protein expression and up-regulation of caspase-3 protein expression.
2012, 19(3):284-288.
DOI: 10.3872/j.issn.1007-385X.2012.3.011
Abstract:
Objective: To investigate the anti-lung cancer effect after infusion of 60Co inactivated-MHC haploidentical cytotoxicity T lymphocytes (CTLs) induced by dendritic cells (DCs). Methods: DCs were cultured from CB6F1 mice and their phenotype was detected by flow cytometry. Mature DC-induced CTLs against metastatic lung cancer cells (Lewis lung cancer, LLC) derived from C57BL/6 mice were cultured in vitro. The LLC tumor bearing C57BL/6 mice were treated with 60Co-inactivated CTL. The anti-lung cancer effect of CTL was identified through cell-mediated cytotoxicity assay. Pathological examination of the liver, spleen, intestinum tenue and skin was performed to observe the graft versus host disease (GVHD) and lung cancer metastasis. The survival time of tumor bearing mice was observed. Results: FCM analysis showed that mature DCs were expanded. The lung weight of LLC metastatic mice treated with inactivated-CTL was decreased obviously (\[0.27±0.06\] vs \[0.52±0.07\] g, P<0.05); and the mean survival time of treated mice prolonged (\[78.10±16.50\] vs \[49.30±6.45\] d, P<0.05). The cell-mediated cytotoxicity of the spleen leukocytes against LLC increased along with the ratio of effector to target (the highest cytotoxicity was \[32.7±1.64\]% when effector〖DK〗∶target =100〖DK〗∶1). There was no obvious GVHD in pathological findings. Conclusion: Inactivated haploidentical CTL can induce antitumor immunological responses and reduce lung cancer metastasis without obvious GVHD.
Objective: To investigate the anti-lung cancer effect after infusion of 60Co inactivated-MHC haploidentical cytotoxicity T lymphocytes (CTLs) induced by dendritic cells (DCs). Methods: DCs were cultured from CB6F1 mice and their phenotype was detected by flow cytometry. Mature DC-induced CTLs against metastatic lung cancer cells (Lewis lung cancer, LLC) derived from C57BL/6 mice were cultured in vitro. The LLC tumor bearing C57BL/6 mice were treated with 60Co-inactivated CTL. The anti-lung cancer effect of CTL was identified through cell-mediated cytotoxicity assay. Pathological examination of the liver, spleen, intestinum tenue and skin was performed to observe the graft versus host disease (GVHD) and lung cancer metastasis. The survival time of tumor bearing mice was observed. Results: FCM analysis showed that mature DCs were expanded. The lung weight of LLC metastatic mice treated with inactivated-CTL was decreased obviously (\[0.27±0.06\] vs \[0.52±0.07\] g, P<0.05); and the mean survival time of treated mice prolonged (\[78.10±16.50\] vs \[49.30±6.45\] d, P<0.05). The cell-mediated cytotoxicity of the spleen leukocytes against LLC increased along with the ratio of effector to target (the highest cytotoxicity was \[32.7±1.64\]% when effector〖DK〗∶target =100〖DK〗∶1). There was no obvious GVHD in pathological findings. Conclusion: Inactivated haploidentical CTL can induce antitumor immunological responses and reduce lung cancer metastasis without obvious GVHD.
2012, 19(3):289-293.
DOI: 10.3872/j.issn.1007-385X.2012.3.012
Abstract:
Objective:To explore the relationship between the expression of G protein-coupled receptor (GPR)54 gene and the biologic behavior or prognosis of ostosarcoma patients.Methods: The expression levels of GPR54 mRNA in 3 types of osteosarcoma cell lines (MG-63, Saos-2 and U-2 OS cells) and one normal osteoblast cell line hF-OB 1.19 were examined using RT-PCR. The expression levels of GPR54 protein in 44 osteosarcoma and 12 benign osteochondroma control paraffin specimen sections were detected by immunohistochemistry. Transwell assay was used to detect the migration and invasion abilities of the 3 types of osteosarcoma cell lines. Results: GPR54mRNA was strongly expressed in all the 3 types of osteosarcoma cell lines (MG-63, Saos-2 and U-2 OS) and the normal osteoblast cell line (\[0.87±0.05\] vs \[088±0.07\], \[0.88±0.08\], \[0.86±0.05\], P＞0.05), but no significant difference existed between each other. Transwell assay displayed a gradually decreased aggressive phenomenon in osteosarcoma cell lines MG-63, Saos-2, U-2 OS (\[18.10±3.21\] vs \[32.40±5.36\], \[61.00±7.15\], P<0.05). The positive expression rate of GPR54 was significantly higher in osteosarcoma than in benign osteochondroma control (81.82% vs 41.67%, P<0.05). Significantly shorter average survival time was displayed in the patients with GPR54 protein positive expression than the in those with GPR54 protein negative expression (\[27.39±6.05\] months vs \[40.33±4.88\] months, P<0.05). Conclusion: The positive expression rate of GPR54 gene is higher in osteosarcoma patients than in benign osteochondroma patients, which might be associated with short survival time in osteosarcoma patients.
Objective:To explore the relationship between the expression of G protein-coupled receptor (GPR)54 gene and the biologic behavior or prognosis of ostosarcoma patients.Methods: The expression levels of GPR54 mRNA in 3 types of osteosarcoma cell lines (MG-63, Saos-2 and U-2 OS cells) and one normal osteoblast cell line hF-OB 1.19 were examined using RT-PCR. The expression levels of GPR54 protein in 44 osteosarcoma and 12 benign osteochondroma control paraffin specimen sections were detected by immunohistochemistry. Transwell assay was used to detect the migration and invasion abilities of the 3 types of osteosarcoma cell lines. Results: GPR54mRNA was strongly expressed in all the 3 types of osteosarcoma cell lines (MG-63, Saos-2 and U-2 OS) and the normal osteoblast cell line (\[0.87±0.05\] vs \[088±0.07\], \[0.88±0.08\], \[0.86±0.05\], P＞0.05), but no significant difference existed between each other. Transwell assay displayed a gradually decreased aggressive phenomenon in osteosarcoma cell lines MG-63, Saos-2, U-2 OS (\[18.10±3.21\] vs \[32.40±5.36\], \[61.00±7.15\], P<0.05). The positive expression rate of GPR54 was significantly higher in osteosarcoma than in benign osteochondroma control (81.82% vs 41.67%, P<0.05). Significantly shorter average survival time was displayed in the patients with GPR54 protein positive expression than the in those with GPR54 protein negative expression (\[27.39±6.05\] months vs \[40.33±4.88\] months, P<0.05). Conclusion: The positive expression rate of GPR54 gene is higher in osteosarcoma patients than in benign osteochondroma patients, which might be associated with short survival time in osteosarcoma patients.
2012, 19(3):294-297.
DOI: 10.3872/j.issn.1007-385X.2012.3.013
Abstract:
Objective: To assess the expressions of CXC chemokine receptor 4 (CXCR4) and CD40 in human cervical carcinoma tissues and evaluate the correlation between their combined expression and the depth of the myometrial invasion. Methods: Fifty-three patients with cervical carcinoma aged of 29-53 years (median 41) from the First Affiliated Hospital of Soochow University were included in this study (2009-2011). The expressions of CXCR4 and CD40 in 53 cases of cervical carcinoma tissues were detected by immunohistochemistry. Then the association between the expressions of these two proteins and the depth of myometrial invasion was analyzed. Results: CXCR4 and CD40 were found to be expressed in cervical carcinoma tissues at different levels, and there was a positive correlation between CD40 and CXCR4 expressions (P=0.035). Of 22 cases which highly expressed CXCR4, 11 were found with a high expression of CD40. Furthermore, combined analysis of CXCR4 and CD40 expressions made results more relevant to the myometrial invasion. The depth of myometrial invasion was significantly higher in patients (90%) with a high expression of both CXCR4 and CD40 than in those (72%) with a high expression of one of the molecules, or those (20%) without high expressions of these two molecules (P<0.05). Conclusion: The expressions of CXCR4 and CD40 in cervical carcinoma tissues are correlated with the depth of the myometrial invasion.
Objective: To assess the expressions of CXC chemokine receptor 4 (CXCR4) and CD40 in human cervical carcinoma tissues and evaluate the correlation between their combined expression and the depth of the myometrial invasion. Methods: Fifty-three patients with cervical carcinoma aged of 29-53 years (median 41) from the First Affiliated Hospital of Soochow University were included in this study (2009-2011). The expressions of CXCR4 and CD40 in 53 cases of cervical carcinoma tissues were detected by immunohistochemistry. Then the association between the expressions of these two proteins and the depth of myometrial invasion was analyzed. Results: CXCR4 and CD40 were found to be expressed in cervical carcinoma tissues at different levels, and there was a positive correlation between CD40 and CXCR4 expressions (P=0.035). Of 22 cases which highly expressed CXCR4, 11 were found with a high expression of CD40. Furthermore, combined analysis of CXCR4 and CD40 expressions made results more relevant to the myometrial invasion. The depth of myometrial invasion was significantly higher in patients (90%) with a high expression of both CXCR4 and CD40 than in those (72%) with a high expression of one of the molecules, or those (20%) without high expressions of these two molecules (P<0.05). Conclusion: The expressions of CXCR4 and CD40 in cervical carcinoma tissues are correlated with the depth of the myometrial invasion.
2012, 19(3):298-302.
DOI: 10.3872/j.issn.1007-385X.2012.3.014
Abstract:
Objective: To investigate the expressions of reversion-inducing cysteine-rich protein with Kazal motifs (RECK) mRNA and matrix metalloproteinase-9 (MMP-9) mRNA in endometrial carcinoma and explore their clinical significance. Methods: Forty-two endometrial carcinoma samples were obtained from gynecological surgery in the Affiliated Hospital of Binzhou Medical University (Mar. 2009 to Dec. 2010). Real-time quantitative PCR was used to assess the expressions of RECK and MMP-9 mRNA in endometrial carcinoma, and their relationship with clinic pathological features of endometrial carcinoma was analyzed. Results: The expression level of RECK mRNA was progressively decreased from the normal to invasive carcinoma (\[6.30±0.34\], \[4.29±0.36\], \[0.24±0.18\]，F=427.35, P<0.05）, while MMP-9 mRNA progressively increased (\[0.08±0.82\], \[5.04±0.30\], \[6.22±0.32\]，F=1117. 52, P<0.05). Both expressions of RECK and MMP-9 mRNA were correlated with TNM stage, histological grade and lymph node metastasis of endometrial carcinoma (P<0.05). There existed a significantly negative correlation between the expressions of RECK mRNA and MMP-9 mRNA in endometrial carcinoma (r=-0.832, P<0.01). Conclusion: Combined detection of RECK mRNA and MMP-9 mRNA by quantitative PCR has a certain value for predicting risk of early diagnosis of endometrial carcinoma and recancerous lesions.
Objective: To investigate the expressions of reversion-inducing cysteine-rich protein with Kazal motifs (RECK) mRNA and matrix metalloproteinase-9 (MMP-9) mRNA in endometrial carcinoma and explore their clinical significance. Methods: Forty-two endometrial carcinoma samples were obtained from gynecological surgery in the Affiliated Hospital of Binzhou Medical University (Mar. 2009 to Dec. 2010). Real-time quantitative PCR was used to assess the expressions of RECK and MMP-9 mRNA in endometrial carcinoma, and their relationship with clinic pathological features of endometrial carcinoma was analyzed. Results: The expression level of RECK mRNA was progressively decreased from the normal to invasive carcinoma (\[6.30±0.34\], \[4.29±0.36\], \[0.24±0.18\]，F=427.35, P<0.05）, while MMP-9 mRNA progressively increased (\[0.08±0.82\], \[5.04±0.30\], \[6.22±0.32\]，F=1117. 52, P<0.05). Both expressions of RECK and MMP-9 mRNA were correlated with TNM stage, histological grade and lymph node metastasis of endometrial carcinoma (P<0.05). There existed a significantly negative correlation between the expressions of RECK mRNA and MMP-9 mRNA in endometrial carcinoma (r=-0.832, P<0.01). Conclusion: Combined detection of RECK mRNA and MMP-9 mRNA by quantitative PCR has a certain value for predicting risk of early diagnosis of endometrial carcinoma and recancerous lesions.
2012, 19(3):303-308.
DOI: 10.3872/j.issn.1007-385X.2012.3.015
Abstract:
Objective:To evaluate the clinical outcome and adverse reaction induced by cetuximab combined with chemotherapy in the treatment of advanced colorectal cancer. Methods: In PubMed, EMBASE, Cochrane Central Register of Controlled Trials (CENTRAL), Chinese Biomedical Abstract Database (CBM) and China National Knowledge Infrastructure Database (CNKI), we searched randomized controlled trials of cetuximab combined with chemotherapy in the treatment of advanced colorectal cancer up to December 2011 and then decided whether the trials met the selection standard and evaluate the quality of these trails. A meta-analysis was performed on the recruited trails. Results: Five reports, including 3 479 advanced colorectal cancer patients, were selected based on our standard. The pooled relative risks (RRs) for overall response rate in all studies were 28.85% in the cetuximab combined with the chemotherapy treatment group and 18.63% in the control chemotherapy group, and the pooled RR \[95% confidence interval (CI)\] was 2.18 (1.24-3.85, P<0.001). The incidence rate of grade 3-4 fatigue was 10.91% in the combined treatment group and 737% in the control group, and the pooled RR (95%CI) was 1.46 (1.19-1.79, P<0.001). The incidence rate of grade 3-4 diarrhea was 20.41% in the combined treatment group and 12.53% in the control group, and the pooled RR (95%CI) was 1.62 (1.37-1.92, P<0.001). Conclusion:Cetuximab combined with chemotherapy obviously elevates the response rate of advanced colorectal cancer patients. However, it also increases the incidence rate of grade 3-4 fatigue and diarrhea.
Objective:To evaluate the clinical outcome and adverse reaction induced by cetuximab combined with chemotherapy in the treatment of advanced colorectal cancer. Methods: In PubMed, EMBASE, Cochrane Central Register of Controlled Trials (CENTRAL), Chinese Biomedical Abstract Database (CBM) and China National Knowledge Infrastructure Database (CNKI), we searched randomized controlled trials of cetuximab combined with chemotherapy in the treatment of advanced colorectal cancer up to December 2011 and then decided whether the trials met the selection standard and evaluate the quality of these trails. A meta-analysis was performed on the recruited trails. Results: Five reports, including 3 479 advanced colorectal cancer patients, were selected based on our standard. The pooled relative risks (RRs) for overall response rate in all studies were 28.85% in the cetuximab combined with the chemotherapy treatment group and 18.63% in the control chemotherapy group, and the pooled RR \[95% confidence interval (CI)\] was 2.18 (1.24-3.85, P<0.001). The incidence rate of grade 3-4 fatigue was 10.91% in the combined treatment group and 737% in the control group, and the pooled RR (95%CI) was 1.46 (1.19-1.79, P<0.001). The incidence rate of grade 3-4 diarrhea was 20.41% in the combined treatment group and 12.53% in the control group, and the pooled RR (95%CI) was 1.62 (1.37-1.92, P<0.001). Conclusion:Cetuximab combined with chemotherapy obviously elevates the response rate of advanced colorectal cancer patients. However, it also increases the incidence rate of grade 3-4 fatigue and diarrhea.
2012, 19(3):309-312.
DOI: 10.3872/j.issn.1007-385X.2012.3.016
Abstract:
目的：观察自体树突状细胞（dendritic cell, DC）联合细胞因子诱导的杀伤细胞（cytokine-induced kill cell, CIK）（DC-CIK）免疫疗法治疗肾癌的近期临床疗效和不良反应，及其对患者外周血淋巴细胞亚群的影响。方法：采集10例肾癌患者外周血单个核细胞，经体外诱导产生DC和CIK细胞，分次回输给患者，随访3~12个月，观察患者临床疗效和不良反应。分别于第1次治疗前1周和每次治疗后1个月取患者外周血，流式细胞仪检测其淋巴细胞亚群变化。结果：本研究中6例晚期肾癌患者接受1~3疗程DC-CIK细胞免疫治疗后，完全缓解1例，部分缓解3例，病情稳定1例，疾病进展1例；近期有效率为60%，临床反应率为90%。治疗前及1~3疗程治疗后患者外周血的CD3+CD4+CD8-、CD3+CD4-CD8+、CD3+CD19-、CD3-CD19+、CD3-CD16+CD56+、CD3+CD16+CD56+、CD3+HLA-DR-、CD3+HLA-DR+、CD3+CD28+CD8+细胞无明显变化（P>0.05），CD4+CD25+T细胞（Treg细胞）治疗后有降低趋势（P＜0.05）。除1例患者出现一过性发热外，余患者无不良反应。结论：DC-CIK免疫治疗肾癌能改善患者免疫抑制状态，提高机体的抗肿瘤免疫效应，有较好的近期疗效，无明显不良反应。
目的：观察自体树突状细胞（dendritic cell, DC）联合细胞因子诱导的杀伤细胞（cytokine-induced kill cell, CIK）（DC-CIK）免疫疗法治疗肾癌的近期临床疗效和不良反应，及其对患者外周血淋巴细胞亚群的影响。方法：采集10例肾癌患者外周血单个核细胞，经体外诱导产生DC和CIK细胞，分次回输给患者，随访3~12个月，观察患者临床疗效和不良反应。分别于第1次治疗前1周和每次治疗后1个月取患者外周血，流式细胞仪检测其淋巴细胞亚群变化。结果：本研究中6例晚期肾癌患者接受1~3疗程DC-CIK细胞免疫治疗后，完全缓解1例，部分缓解3例，病情稳定1例，疾病进展1例；近期有效率为60%，临床反应率为90%。治疗前及1~3疗程治疗后患者外周血的CD3+CD4+CD8-、CD3+CD4-CD8+、CD3+CD19-、CD3-CD19+、CD3-CD16+CD56+、CD3+CD16+CD56+、CD3+HLA-DR-、CD3+HLA-DR+、CD3+CD28+CD8+细胞无明显变化（P>0.05），CD4+CD25+T细胞（Treg细胞）治疗后有降低趋势（P＜0.05）。除1例患者出现一过性发热外，余患者无不良反应。结论：DC-CIK免疫治疗肾癌能改善患者免疫抑制状态，提高机体的抗肿瘤免疫效应，有较好的近期疗效，无明显不良反应。
2012, 19(3):313-316.
DOI: 10.3872/j.issn.1007-385X.2012.3.017
Abstract:
目的：探讨携带癌胚抗原（carcinoembryonic antigen，CEA）基因重组腺相关病毒rAAV/CEA感染直肠癌患者的DC诱导CTL对肿瘤的杀伤作用。方法:抽取直肠癌患者的外周血培养DC细胞，用rAAV/CEA感染DC，DC成熟后与淋巴细胞混合诱导产生CTL，用流式细胞仪分别检测感染和未感染病毒的DC的表型及其诱导的CTL杀伤活性。结果：实验组DC的CD40、CD80、CD86表达显著增高\[(81.95±1.45)% vs (72.27±1.59)% ，( 80.10±1.03)% vs ( 70.59±1.04)%，(58.93±1.84)% vs ( 48.66±1.97)%；均P<0.05\]。实验组DC中CEA的表达率显著增高\[(59.04±1.68)% vs (46.03±1.23)%；均P<0.05\]。在效靶比为12.5∶1、25∶1、50∶1、100∶1时，实验组CTL对直肠癌LoVo细胞的杀伤率明显提高\[(12.65±030)% vs (6.59±0.21)%，(25.77±2.53)% vs (8.35±112)%，(43.36±0.83)% vs (12.79±0.23)% ，(61.03±183)% vs (23.35±0.69)%；均P<0.05\]，并且随着效应细胞比例的增加，杀伤率也逐渐增加。结论：携带CEA重组腺相关病毒感染DC激活CTL对肿瘤细胞能产生高效的杀伤作用。
目的：探讨携带癌胚抗原（carcinoembryonic antigen，CEA）基因重组腺相关病毒rAAV/CEA感染直肠癌患者的DC诱导CTL对肿瘤的杀伤作用。方法:抽取直肠癌患者的外周血培养DC细胞，用rAAV/CEA感染DC，DC成熟后与淋巴细胞混合诱导产生CTL，用流式细胞仪分别检测感染和未感染病毒的DC的表型及其诱导的CTL杀伤活性。结果：实验组DC的CD40、CD80、CD86表达显著增高\[(81.95±1.45)% vs (72.27±1.59)% ，( 80.10±1.03)% vs ( 70.59±1.04)%，(58.93±1.84)% vs ( 48.66±1.97)%；均P<0.05\]。实验组DC中CEA的表达率显著增高\[(59.04±1.68)% vs (46.03±1.23)%；均P<0.05\]。在效靶比为12.5∶1、25∶1、50∶1、100∶1时，实验组CTL对直肠癌LoVo细胞的杀伤率明显提高\[(12.65±030)% vs (6.59±0.21)%，(25.77±2.53)% vs (8.35±112)%，(43.36±0.83)% vs (12.79±0.23)% ，(61.03±183)% vs (23.35±0.69)%；均P<0.05\]，并且随着效应细胞比例的增加，杀伤率也逐渐增加。结论：携带CEA重组腺相关病毒感染DC激活CTL对肿瘤细胞能产生高效的杀伤作用。
2012, 19(3):317-319.
DOI: 10.3872/j.issn.1007-385X.2012.3.018
Abstract:
目的：采用细胞因子诱导的杀伤细胞（cytokine induced killer cell，CIK cell）培养基（含低浓度IL-2、IFN-γ、IL-12和anti-CD3）和NK培养基（含高浓度IL-2和anti-CD3）体外刺激诱导CIK，分析CIK细胞的扩增、表型和细胞因子分泌情况。方法：健康自愿者来源的外周血单个核细胞（peripheral blood mononuclear cell，PBMC），分别加入CIK培养基和NK培养基进行诱导培养，24 h后，换用含IL-2的1%自体血浆培养基继续诱导培养2周。在培养的第6天和第10天检测培养上清中IFN-γ的分泌情况，在培养的第10天以流式细胞术检测两组培养细胞表面CD3、CD4、CD8、CD16、CD56的表达情况。结果：CIK培养基和NK培养基均可使培养细胞在第7天时开始明显扩增，第10天时可扩增至200倍。其中，CIK培养基获得的细胞扩增倍数明显高于NK培养基，其IFN-γ分泌明显高于NK培养基组\[（724.7±434.8） vs（108.9±60.6）pg/ml，P<0.01\]；而以NK培养基培养的细胞，CD3-CD16+CD56+细胞比例高于CIK培养基\[（6.27±3.33)% vs (2.88±1.88)%，P<0.05\]。结论：不同成分的CIK培养基和NK培养基均可促进CIK细胞的扩增和成熟，但呈现不同的生物学特征。
目的：采用细胞因子诱导的杀伤细胞（cytokine induced killer cell，CIK cell）培养基（含低浓度IL-2、IFN-γ、IL-12和anti-CD3）和NK培养基（含高浓度IL-2和anti-CD3）体外刺激诱导CIK，分析CIK细胞的扩增、表型和细胞因子分泌情况。方法：健康自愿者来源的外周血单个核细胞（peripheral blood mononuclear cell，PBMC），分别加入CIK培养基和NK培养基进行诱导培养，24 h后，换用含IL-2的1%自体血浆培养基继续诱导培养2周。在培养的第6天和第10天检测培养上清中IFN-γ的分泌情况，在培养的第10天以流式细胞术检测两组培养细胞表面CD3、CD4、CD8、CD16、CD56的表达情况。结果：CIK培养基和NK培养基均可使培养细胞在第7天时开始明显扩增，第10天时可扩增至200倍。其中，CIK培养基获得的细胞扩增倍数明显高于NK培养基，其IFN-γ分泌明显高于NK培养基组\[（724.7±434.8） vs（108.9±60.6）pg/ml，P<0.01\]；而以NK培养基培养的细胞，CD3-CD16+CD56+细胞比例高于CIK培养基\[（6.27±3.33)% vs (2.88±1.88)%，P<0.05\]。结论：不同成分的CIK培养基和NK培养基均可促进CIK细胞的扩增和成熟，但呈现不同的生物学特征。
2012, 19(3):320-325.
DOI: 10.3872/j.issn.1007-385X.2012.3.019
Abstract:
[摘 要] 髓源抑制性细胞（myeloid-derived suppressor cell，MDSC）是肿瘤和其他多种病理条件下免疫抑制网络的主要成员之一，在肿瘤的发生、发展及肿瘤逃逸过程中发挥重要作用。在自身免疫疾病、感染、肿瘤等病理情况下，髓系细胞分化发生异常，导致MDSC累积，此过程受到髓系细胞分化发育相关的多条信号通路的调控和多种细胞因子的影响。多种细胞因子或转录因子都通过JAK-STAT和NF-κB信号通路来影响髓系细胞的分化和成熟，进而影响MDSC的产生和活化。STAT蛋白在调控MDSC的产生、扩增和功能方面发挥着重要作用，NF-κB可能提供MDSC累积的第二条途径。配对免疫球蛋白样受体-B（paired immunoglobin-like receptor-B，PIR-B）参与调控了MDSC的功能和分化，PIR-A和PIR-B是调控MDSC分化为M1或M2型巨噬细胞的关键因子，同时对MDSC的免疫抑制功能和促进肿瘤发展有重要作用。Notch通路的异常也导致了MDSC的累积。C/EBP-β、IRF-8、HIF-1α等转录因子在调控MDSC产生、累积和功能方面也有重要的作用。
[摘 要] 髓源抑制性细胞（myeloid-derived suppressor cell，MDSC）是肿瘤和其他多种病理条件下免疫抑制网络的主要成员之一，在肿瘤的发生、发展及肿瘤逃逸过程中发挥重要作用。在自身免疫疾病、感染、肿瘤等病理情况下，髓系细胞分化发生异常，导致MDSC累积，此过程受到髓系细胞分化发育相关的多条信号通路的调控和多种细胞因子的影响。多种细胞因子或转录因子都通过JAK-STAT和NF-κB信号通路来影响髓系细胞的分化和成熟，进而影响MDSC的产生和活化。STAT蛋白在调控MDSC的产生、扩增和功能方面发挥着重要作用，NF-κB可能提供MDSC累积的第二条途径。配对免疫球蛋白样受体-B（paired immunoglobin-like receptor-B，PIR-B）参与调控了MDSC的功能和分化，PIR-A和PIR-B是调控MDSC分化为M1或M2型巨噬细胞的关键因子，同时对MDSC的免疫抑制功能和促进肿瘤发展有重要作用。Notch通路的异常也导致了MDSC的累积。C/EBP-β、IRF-8、HIF-1α等转录因子在调控MDSC产生、累积和功能方面也有重要的作用。
2012, 19(3):326-329.
DOI: 10.3872/j.issn.1007-385X.2012.3.020
Abstract:
[摘 要] 肺癌干细胞（lung cancer stem cell，LCSC）是肺癌组织中一小群具有自我更新及分化潜能，并且具有耐药性的特殊细胞。LCSC是目前肺癌研究中的热点。研究者通过对肺癌中CD133阳性细胞侧群（side population，SP）细胞及耐药存活细胞（drug surviving cell，DSC）的研究，证实了LCSC的存在；而乙醛脱氢酶1（aldehyde dehydrogenase 1，ALDH1）、八聚体结合转录因子-4（octamer-binding transcription factor 4，Oct-4）及CD44可能是鉴定LCSC的新标志物。Notch、Wnt及Hedgehog信号通路与LCSC的发生及维持关系密切，靶向阻断这些信号通路可以减少肺癌组织中LCSC的比例，可能成为肺癌靶向治疗的新策略
[摘 要] 肺癌干细胞（lung cancer stem cell，LCSC）是肺癌组织中一小群具有自我更新及分化潜能，并且具有耐药性的特殊细胞。LCSC是目前肺癌研究中的热点。研究者通过对肺癌中CD133阳性细胞侧群（side population，SP）细胞及耐药存活细胞（drug surviving cell，DSC）的研究，证实了LCSC的存在；而乙醛脱氢酶1（aldehyde dehydrogenase 1，ALDH1）、八聚体结合转录因子-4（octamer-binding transcription factor 4，Oct-4）及CD44可能是鉴定LCSC的新标志物。Notch、Wnt及Hedgehog信号通路与LCSC的发生及维持关系密切，靶向阻断这些信号通路可以减少肺癌组织中LCSC的比例，可能成为肺癌靶向治疗的新策略
2012, 19(3):330-335.
DOI: 10.3872/j.issn.1007-385X.2012.3.021
Abstract:
[摘 要] 树突状细胞（dendritic cell，DC）是目前已知功能最强的抗原提呈细胞。一种安全可行的肿瘤免疫基因治疗方法是将肿瘤相关抗原的mRNA或肿瘤细胞总RNA转染DC，使之将肿瘤抗原信息提呈给T细胞，产生特异性免疫应答。目前，临床研究较多的是髓系DC，体外培养DC的技术也一直在不断地改进。RNA转染DC的方法主要有脂质体介导的转染、裸RNA转染（被动转染）、电穿孔转染和核转染法。动物实验已经证实，RNA负载的DC能有效地诱导抗原特异性免疫应答。近年来研究者们对RNA修饰的DC瘤苗治疗临床常见各种肿瘤的免疫学效应进行了广泛的临床前和临床试验研究，已显示出该类疫苗具有广阔的应用前景。
[摘 要] 树突状细胞（dendritic cell，DC）是目前已知功能最强的抗原提呈细胞。一种安全可行的肿瘤免疫基因治疗方法是将肿瘤相关抗原的mRNA或肿瘤细胞总RNA转染DC，使之将肿瘤抗原信息提呈给T细胞，产生特异性免疫应答。目前，临床研究较多的是髓系DC，体外培养DC的技术也一直在不断地改进。RNA转染DC的方法主要有脂质体介导的转染、裸RNA转染（被动转染）、电穿孔转染和核转染法。动物实验已经证实，RNA负载的DC能有效地诱导抗原特异性免疫应答。近年来研究者们对RNA修饰的DC瘤苗治疗临床常见各种肿瘤的免疫学效应进行了广泛的临床前和临床试验研究，已显示出该类疫苗具有广阔的应用前景。
2012, 19(3):336-340.
DOI: 10.3872/j.issn.1007-385X.2012.3.022
Abstract:
[摘 要] 树突状细胞（dendritic cell，DC）作为体内功能最强的专职抗原提呈细胞，广泛分布于各种组织器官中，在激活肿瘤特异性免疫中发挥重要作用。黑素瘤相关抗原3（melanoma-associated antigen 3，MAGE-3）是一种肿瘤-睾丸抗原（cancer-testis antigen，CTA），属于黑素瘤相关抗原家族（melanoma-associated antigen family，MAGE family），在上皮细胞来源的多种肿瘤细胞表面都有不同程度的表达。MAGE家族类肿瘤表面标志物能被用于早期发现肿瘤细胞，并针对该类抗原进行特异性的免疫治疗，是肿瘤免疫治疗的理想靶抗原。由于DC能将肿瘤相关性抗原提呈给T淋巴细胞，产生抗原特异性免疫反应，因此DC肿瘤疫苗为肿瘤免疫治疗提供了一种有效手段。目前，针对MAGE-3抗原的DC肿瘤疫苗研究已经广泛开展，可以通过多种不同的方式将MAGE-3负载DC，并取得了一定的临床疗效，为DC肿瘤疫苗的临床应用带来了曙光。
[摘 要] 树突状细胞（dendritic cell，DC）作为体内功能最强的专职抗原提呈细胞，广泛分布于各种组织器官中，在激活肿瘤特异性免疫中发挥重要作用。黑素瘤相关抗原3（melanoma-associated antigen 3，MAGE-3）是一种肿瘤-睾丸抗原（cancer-testis antigen，CTA），属于黑素瘤相关抗原家族（melanoma-associated antigen family，MAGE family），在上皮细胞来源的多种肿瘤细胞表面都有不同程度的表达。MAGE家族类肿瘤表面标志物能被用于早期发现肿瘤细胞，并针对该类抗原进行特异性的免疫治疗，是肿瘤免疫治疗的理想靶抗原。由于DC能将肿瘤相关性抗原提呈给T淋巴细胞，产生抗原特异性免疫反应，因此DC肿瘤疫苗为肿瘤免疫治疗提供了一种有效手段。目前，针对MAGE-3抗原的DC肿瘤疫苗研究已经广泛开展，可以通过多种不同的方式将MAGE-3负载DC，并取得了一定的临床疗效，为DC肿瘤疫苗的临床应用带来了曙光。
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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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- 网址:http://www.biother.cn
- 刊号：ISSN 1007-385X
- CN 31-1725/R
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.