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Volume 19,Issue 1,2012 Table of Contents
2012, 19(1):1-10.
DOI: 10.3872/j.issn.1007-385X.2012.1.001
Abstract:
Dendritic cell (DC)-based cancer vaccines have been studied for several decades and a line of clinical trials has been completed or in process since Ralph M. Steinman, a Nobel Prize winner in Physiology or Medicine 2011, found DCs and their crucial role in adaptive immunity. Although three DC-based cancer vaccines (Sipuleucel-T, CreaVax RCC and Hybricell) have been so far approved for marketing, the DC-based immunotherapy has not yet been approved as a standard treatment for cancer. In order to make domestic peers comprehensively understood the current status of global clinical trials of DC vaccines in cancer, the general situation (regional and country distribution, involved cancer type, start year and clinical trial phase) of these clinical trials is introduced and 43 completed and published clinical trials (selecting subject, DC culturing method, vaccination schedule, effectiveness assessment method and trial result) are summarized on the basis of clinical trial registry websites accepted by International Committee of Medical Journal Editors ( ICMJE ) and PubMed web databases. Then the current development trend and existing problems of clinical trials using DC-based cancer vaccines are emphatically analyzed. Finally, several proposals to improve the clinical studies with DC vaccines against cancer are put forward as follows: perfection and refinement of our regulatory policy on clinical trials of DC-based cancer vaccines; close concern of international “in vivo DC targeting” strategy; grasping to establish DC culturing method, vaccination schedule and effect assessment standard; strengthening quality monitoring of DC-based cancer vaccines; attention to basic research and clinical trial registry of DC-based vaccines for cancer; improving the clinical trial protocols of DC-based cancer vaccines; and careful selection of subjects, effect assessment time points and synchronous application of immune monitoring during cancer treatment. The authors wish to arouse domestic peers’ attention and stimulate discussion between them, and these problems will be studied and addressed as possible as we can in the future.
Dendritic cell (DC)-based cancer vaccines have been studied for several decades and a line of clinical trials has been completed or in process since Ralph M. Steinman, a Nobel Prize winner in Physiology or Medicine 2011, found DCs and their crucial role in adaptive immunity. Although three DC-based cancer vaccines (Sipuleucel-T, CreaVax RCC and Hybricell) have been so far approved for marketing, the DC-based immunotherapy has not yet been approved as a standard treatment for cancer. In order to make domestic peers comprehensively understood the current status of global clinical trials of DC vaccines in cancer, the general situation (regional and country distribution, involved cancer type, start year and clinical trial phase) of these clinical trials is introduced and 43 completed and published clinical trials (selecting subject, DC culturing method, vaccination schedule, effectiveness assessment method and trial result) are summarized on the basis of clinical trial registry websites accepted by International Committee of Medical Journal Editors ( ICMJE ) and PubMed web databases. Then the current development trend and existing problems of clinical trials using DC-based cancer vaccines are emphatically analyzed. Finally, several proposals to improve the clinical studies with DC vaccines against cancer are put forward as follows: perfection and refinement of our regulatory policy on clinical trials of DC-based cancer vaccines; close concern of international “in vivo DC targeting” strategy; grasping to establish DC culturing method, vaccination schedule and effect assessment standard; strengthening quality monitoring of DC-based cancer vaccines; attention to basic research and clinical trial registry of DC-based vaccines for cancer; improving the clinical trial protocols of DC-based cancer vaccines; and careful selection of subjects, effect assessment time points and synchronous application of immune monitoring during cancer treatment. The authors wish to arouse domestic peers’ attention and stimulate discussion between them, and these problems will be studied and addressed as possible as we can in the future.
2012, 19(1):11-16.
DOI: 10.3872/j.issn.1007-385X.2012.1.002
Abstract:
Objective : To observe the effect and mechanism of IL-27 on the morphology and function of human peripheral blood mononuclear cell (PBMC)-derived dendritic cells (DCs). Methods: PBMCs were purified from peripheral blood of healthy adults and incubated with granulocyte/macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 7 days. The DCs were divided into four groups by different stimulatory factors at the fifth day: a negative group, a positive group (TNF-α: 20 ng/ml), IL-27 group (20 ng/ml) and TNF-α+IL-27 group (co-cytokines group, 10 ng/ml TNF-α+ IL-27 10 ng/ml). The morphology of DCs was observed under an inverted microscope at the 7th day. The expressions of CD1a and CD83, CD80 and CD86 on DCs were analyzed by flow cytometry. The mRNA levels of chemokine receptors of CCR5 and CCR7 on DCs were analyzed by RT-PCR.The stimulatory ability of DCs on proliferation of allogeneic T cells was detected by MLR. The proteins of P-STAT1 and P-STAT3 were detected by Western blotting. Results : DCs from the IL-27 group and TNF-α + IL-27 group had a typical morphological characteristic of mature DCs after 7 days. The expressions of co-stimulatory molecules of CD1a and CD83 (\[35.75±410\]%, \[ 52.49± 2.65\]% vs \[2329±449\]%, P<0.05), CD80 and CD86 (\[39.06±1.61\]%, \[54.10±0.46\]% vs \[22.66±3.20\]%, P<0.05), chemokine receptor of CCR7 (3.98±0.09, 4.75±0.11 vs 3.09±0.18, P<0.05) and the proteins of P-STAT1/STAT3 on DCs were up-regulated both in the IL-27 group and co-cytokines group compared with the negative group, while chemokine receptor of CCR5 (0.99±0.03, 0.61±0.02 vs 1.23±0.26, P<0.05) was down-regulated. The proliferation of T cells was improved by DCs from the IL-27 group and TNF-α + IL-27 group. The tendency was especially obvious in the co-cytokines group. Conclusion : IL-27 can directly or synergically enhance the differentiation, maturation and antigen presentation ability of DCs with TNF-α. The mechanism might relate to the activation of P-STAT1/STAT3 signal transduction pathway.
Objective : To observe the effect and mechanism of IL-27 on the morphology and function of human peripheral blood mononuclear cell (PBMC)-derived dendritic cells (DCs). Methods: PBMCs were purified from peripheral blood of healthy adults and incubated with granulocyte/macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 7 days. The DCs were divided into four groups by different stimulatory factors at the fifth day: a negative group, a positive group (TNF-α: 20 ng/ml), IL-27 group (20 ng/ml) and TNF-α+IL-27 group (co-cytokines group, 10 ng/ml TNF-α+ IL-27 10 ng/ml). The morphology of DCs was observed under an inverted microscope at the 7th day. The expressions of CD1a and CD83, CD80 and CD86 on DCs were analyzed by flow cytometry. The mRNA levels of chemokine receptors of CCR5 and CCR7 on DCs were analyzed by RT-PCR.The stimulatory ability of DCs on proliferation of allogeneic T cells was detected by MLR. The proteins of P-STAT1 and P-STAT3 were detected by Western blotting. Results : DCs from the IL-27 group and TNF-α + IL-27 group had a typical morphological characteristic of mature DCs after 7 days. The expressions of co-stimulatory molecules of CD1a and CD83 (\[35.75±410\]%, \[ 52.49± 2.65\]% vs \[2329±449\]%, P<0.05), CD80 and CD86 (\[39.06±1.61\]%, \[54.10±0.46\]% vs \[22.66±3.20\]%, P<0.05), chemokine receptor of CCR7 (3.98±0.09, 4.75±0.11 vs 3.09±0.18, P<0.05) and the proteins of P-STAT1/STAT3 on DCs were up-regulated both in the IL-27 group and co-cytokines group compared with the negative group, while chemokine receptor of CCR5 (0.99±0.03, 0.61±0.02 vs 1.23±0.26, P<0.05) was down-regulated. The proliferation of T cells was improved by DCs from the IL-27 group and TNF-α + IL-27 group. The tendency was especially obvious in the co-cytokines group. Conclusion : IL-27 can directly or synergically enhance the differentiation, maturation and antigen presentation ability of DCs with TNF-α. The mechanism might relate to the activation of P-STAT1/STAT3 signal transduction pathway.
2012, 19(1):17-21.
DOI: 10.3872/j.issn.1007-385X.2012.1.003
Abstract:
Objective : To observe the effect of stimulation with combined IL-12 and IL-18 on exosomes secreted by dendritic cells （DCs）, and so as to facilitate exploring high-efficiency DC derived exosome(Dex) cancer vaccines. Methods: DCs induced from human peripheral blood mononuclear cells (PBMCs) were stimulated with IL-12+IL-18，IL-12 or IL-18 for the combined group，IL-12 group and IL-18 group respectively; and no stimulant for the control group. Dex was extracted from DCs. The expressions of HLA-DR and CD83 in Dex were detected by Western blotting. CD54, CD80 and CD86 expressions were determined by flow cytometry. The T cell proliferation stimulated by Dex was detected with MTT method. IFN-γ in the T cell supernatant was detected by ELISA. Results: HLA-DR and CD83 proteins were expressed in Dex of all the four groups. The molecule expressions for combined group were CD54( 32367± 44.06 vs 246.17±31.91, 236.33±33.87, 167.67±28.73, P<0.05), CD80（406.37±39.18 vs 331.67±3615, 335.67±41.38, 260.00±35.58, P<0.05), and CD86（390.50±38.06 vs 314.33±36.64, 319.00± 33.10, 246.83± 30.55, P<0.05), which were higher than those in the IL-12 group, IL-18 group and control group. The T cell proliferation in the combined group （1.98±0.31 vs 1.55±0.23, 1.57±0.21, 1.10±0.18, P<005) was higher than that in the IL-12 group, IL-18 group and control group. IFN-γ secreted by T cells in the combined group （436.67±61.8 vs 295.04±40.25, 358.18±55.77, 225.00±36.44, P<0.05) was higher than that in the IL-12 group, IL-18 group and control group. Conclusion: Stimulation of DCs with combined IL-12 and IL-18 can enhance Dex express CD54, CD80 and CD86, promote Dex-induced T cell proliferation and increase IFN-γ secretion by T cells.
Objective : To observe the effect of stimulation with combined IL-12 and IL-18 on exosomes secreted by dendritic cells （DCs）, and so as to facilitate exploring high-efficiency DC derived exosome(Dex) cancer vaccines. Methods: DCs induced from human peripheral blood mononuclear cells (PBMCs) were stimulated with IL-12+IL-18，IL-12 or IL-18 for the combined group，IL-12 group and IL-18 group respectively; and no stimulant for the control group. Dex was extracted from DCs. The expressions of HLA-DR and CD83 in Dex were detected by Western blotting. CD54, CD80 and CD86 expressions were determined by flow cytometry. The T cell proliferation stimulated by Dex was detected with MTT method. IFN-γ in the T cell supernatant was detected by ELISA. Results: HLA-DR and CD83 proteins were expressed in Dex of all the four groups. The molecule expressions for combined group were CD54( 32367± 44.06 vs 246.17±31.91, 236.33±33.87, 167.67±28.73, P<0.05), CD80（406.37±39.18 vs 331.67±3615, 335.67±41.38, 260.00±35.58, P<0.05), and CD86（390.50±38.06 vs 314.33±36.64, 319.00± 33.10, 246.83± 30.55, P<0.05), which were higher than those in the IL-12 group, IL-18 group and control group. The T cell proliferation in the combined group （1.98±0.31 vs 1.55±0.23, 1.57±0.21, 1.10±0.18, P<005) was higher than that in the IL-12 group, IL-18 group and control group. IFN-γ secreted by T cells in the combined group （436.67±61.8 vs 295.04±40.25, 358.18±55.77, 225.00±36.44, P<0.05) was higher than that in the IL-12 group, IL-18 group and control group. Conclusion: Stimulation of DCs with combined IL-12 and IL-18 can enhance Dex express CD54, CD80 and CD86, promote Dex-induced T cell proliferation and increase IFN-γ secretion by T cells.
2012, 19(1):22-28.
DOI: 10.3872/j.issn.1007-385X.2012.1.004
Abstract:
Objective : To observe changes in immune function of dendritic cells(DCs) infected by recombinant adenovirus containing human prostate specific membrane antigen (PSMA) and 4-1BB ligand [STBX〗 (4-1BBL)〖STBZ〗 genes. Methods: The empty Ad-GFP infected immature DCs at different multiplicities of infection (MOI), and the optimal MOI was determined. DCs from peripheral blood of healthy volunteers were infected with recombinant adenoviruses at the optimal MOI. The DCs were allocated into five groups: co-infected group (Ad-PSMA/4-1BBL-DC), PSMA-infected group (Ad-PSMA-DC), 4-1BBL-infected group (Ad-4-1BBL-DC), negative control group (Ad-GFP-DC) and non-infected group. Morphological changes of DCs were observed under a fluorescence microscope; and the phenotype changes of CD80, CD83, CD86 of tranfected DCs were detected by FACS, and the expressions of PSMA and 4-1BBL proteins was detected by Western blotting. IL-12 concentrations of DC supernatant in different groups were measured by ELISA. Allogenic T cell proliferations stimulated with DCs in different groups and the cytotoxic effect on prostate cancer cell lines LNCap, Du145 and 22RV were analyzed by CCK-8 assay. Results: The optimal MOI was 200. The immunophenotype expressed in the co-infected DC group, such as CD80, CD83, CD86, HLA-DR, was significantly higher than that expressed in the other DC groups (\[38.72±0.99\]%，\[44.65±0.77\]%，\[63.60±0.75\]%，\[62.25±0.58\]% vs \[28.27±104\]%，\[ 2808± 116%\]，\[41.05±1.33\]%，\[46.87±1.12\]%； P<0.05). The secretion of IL-12 in supernatant was greatly higher in the co-infected group than that in single-infected group or non-infected DC group (\[134.29±222\] vs \[ 79.51± 160\], \[70.33±1.13\], \[69.67±1.43\], \[28.88±2.97\] pg；P<0.05). At the same rate of DCs to T, the co-infected group had a more significant increase in allogenic T cell proliferation than other single-infected group or non-infected group (P<0.05). The cytotoxic rate of PSMA/4-1BBL-DC-CTL against PSMA positive target LNCap cells was significantly stronger than that against PSMA negative target cells such as DU145 and 22RV cells (P<0.05), and also higher than that of PSMA-DC-CTL and 4-1BBL-DC-CTL (P<0.05). Conclusion: Ad-PSMA/4-1BBL-infected DCs not only own high secretion of IL-12, but also stimulate and enhance the cytotoxic effect of tumor specific CTLs against PSMA positive prostate cancer cells which highly express PSMA protein. In addition, the DCs infected with Ad-PSMA and Ad-4-1BBL induce more effective tumor specific CTLs than single-infected DCs.
Objective : To observe changes in immune function of dendritic cells(DCs) infected by recombinant adenovirus containing human prostate specific membrane antigen (PSMA) and 4-1BB ligand [STBX〗 (4-1BBL)〖STBZ〗 genes. Methods: The empty Ad-GFP infected immature DCs at different multiplicities of infection (MOI), and the optimal MOI was determined. DCs from peripheral blood of healthy volunteers were infected with recombinant adenoviruses at the optimal MOI. The DCs were allocated into five groups: co-infected group (Ad-PSMA/4-1BBL-DC), PSMA-infected group (Ad-PSMA-DC), 4-1BBL-infected group (Ad-4-1BBL-DC), negative control group (Ad-GFP-DC) and non-infected group. Morphological changes of DCs were observed under a fluorescence microscope; and the phenotype changes of CD80, CD83, CD86 of tranfected DCs were detected by FACS, and the expressions of PSMA and 4-1BBL proteins was detected by Western blotting. IL-12 concentrations of DC supernatant in different groups were measured by ELISA. Allogenic T cell proliferations stimulated with DCs in different groups and the cytotoxic effect on prostate cancer cell lines LNCap, Du145 and 22RV were analyzed by CCK-8 assay. Results: The optimal MOI was 200. The immunophenotype expressed in the co-infected DC group, such as CD80, CD83, CD86, HLA-DR, was significantly higher than that expressed in the other DC groups (\[38.72±0.99\]%，\[44.65±0.77\]%，\[63.60±0.75\]%，\[62.25±0.58\]% vs \[28.27±104\]%，\[ 2808± 116%\]，\[41.05±1.33\]%，\[46.87±1.12\]%； P<0.05). The secretion of IL-12 in supernatant was greatly higher in the co-infected group than that in single-infected group or non-infected DC group (\[134.29±222\] vs \[ 79.51± 160\], \[70.33±1.13\], \[69.67±1.43\], \[28.88±2.97\] pg；P<0.05). At the same rate of DCs to T, the co-infected group had a more significant increase in allogenic T cell proliferation than other single-infected group or non-infected group (P<0.05). The cytotoxic rate of PSMA/4-1BBL-DC-CTL against PSMA positive target LNCap cells was significantly stronger than that against PSMA negative target cells such as DU145 and 22RV cells (P<0.05), and also higher than that of PSMA-DC-CTL and 4-1BBL-DC-CTL (P<0.05). Conclusion: Ad-PSMA/4-1BBL-infected DCs not only own high secretion of IL-12, but also stimulate and enhance the cytotoxic effect of tumor specific CTLs against PSMA positive prostate cancer cells which highly express PSMA protein. In addition, the DCs infected with Ad-PSMA and Ad-4-1BBL induce more effective tumor specific CTLs than single-infected DCs.
2012, 19(1):29-34.
DOI: 10.3872/j.issn.1007-385X.2012.1.005
Abstract:
Objective : To explore the potential of autologous dendritic cells (DCs) pulsed with HER-2/neu peptide in inducing specific cytotoxic T lymphocyte (CTL) response and feasibility of breast cancer vaccines. Methods: Seventeen breast cancer patients with positive HLA-A201 and HER-2/neu were enrolled and their peripheral blood mononuclear cells and lymphocytes were isolated and induced into DCs and pulsed with HER-2/neu peptide. The killing effect of CTLs against T2 cell line pulsed with HLA-A201-binding peptide HER-2/neu was determined. The patients were inoculated subcutaneously near the inguinal region with auto-DCs pulsed with HER-2/neu peptide for 4 times every week. The immunological responses and clinical responses were examined in 1 week after the final vaccination. Results: The average percentage of special CTLs primed by DCs pulsed with HER-2/neu peptide was significantly higher than that in the control group (CTLs primed by DCs unloaded with HER-2/neu peptide) （\[5.41±1.44\]% vs \[0.41±0.12\]%, P<0.05). CTLs induced by DCs exerted a stronger killing effect on T2 cell line pulsed with HER-2/neu peptide than that in control group （\[35.5±4.7\]% vs \[11.2±1.4\]% at the ratio of E \[effect\] to T \[target\] as 30 ∶1，P<0.05). Vaccination of DCs was well tolerated and no toxicity was observed. The cytokine levels in sera such as IL-2, IL-12 and IFN-γ were increased after vaccinations （\[148.79±28.32\] ng/ml vs \[409.09±89.39\] ng/ml,\[24.49±56.23\] ng/ml vs \[56.23±14.08\] ng/ml, \[67.77±39.35\] ng/ml vs \[146.57±25.97\] ng/ml, respectively, all P<0.05). The cytokine levels in sera such as TNF-α and IL-10 had no significant changes before and after vaccination. The results of DTH test were positive in 8 patients (8/17), and the percentages of antigen-specific IFN-γ + CD8 +T increased in 8 patients (8/17). Conclusion: Auto-DC vaccines pulsed with HER-2/neu peptide can elicit specific immune responses ex vivo and in vivo, and induce secretion of Th1 type cytokines from DCs and have no adverse reaction.
Objective : To explore the potential of autologous dendritic cells (DCs) pulsed with HER-2/neu peptide in inducing specific cytotoxic T lymphocyte (CTL) response and feasibility of breast cancer vaccines. Methods: Seventeen breast cancer patients with positive HLA-A201 and HER-2/neu were enrolled and their peripheral blood mononuclear cells and lymphocytes were isolated and induced into DCs and pulsed with HER-2/neu peptide. The killing effect of CTLs against T2 cell line pulsed with HLA-A201-binding peptide HER-2/neu was determined. The patients were inoculated subcutaneously near the inguinal region with auto-DCs pulsed with HER-2/neu peptide for 4 times every week. The immunological responses and clinical responses were examined in 1 week after the final vaccination. Results: The average percentage of special CTLs primed by DCs pulsed with HER-2/neu peptide was significantly higher than that in the control group (CTLs primed by DCs unloaded with HER-2/neu peptide) （\[5.41±1.44\]% vs \[0.41±0.12\]%, P<0.05). CTLs induced by DCs exerted a stronger killing effect on T2 cell line pulsed with HER-2/neu peptide than that in control group （\[35.5±4.7\]% vs \[11.2±1.4\]% at the ratio of E \[effect\] to T \[target\] as 30 ∶1，P<0.05). Vaccination of DCs was well tolerated and no toxicity was observed. The cytokine levels in sera such as IL-2, IL-12 and IFN-γ were increased after vaccinations （\[148.79±28.32\] ng/ml vs \[409.09±89.39\] ng/ml,\[24.49±56.23\] ng/ml vs \[56.23±14.08\] ng/ml, \[67.77±39.35\] ng/ml vs \[146.57±25.97\] ng/ml, respectively, all P<0.05). The cytokine levels in sera such as TNF-α and IL-10 had no significant changes before and after vaccination. The results of DTH test were positive in 8 patients (8/17), and the percentages of antigen-specific IFN-γ + CD8 +T increased in 8 patients (8/17). Conclusion: Auto-DC vaccines pulsed with HER-2/neu peptide can elicit specific immune responses ex vivo and in vivo, and induce secretion of Th1 type cytokines from DCs and have no adverse reaction.
2012, 19(1):35-39.
DOI: 10.3872/j.issn.1007-385X.2012.1.006
Abstract:
Objective : To explore the cytotoxic effect of cyctotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) modified by EPHA2 gene on U251 glioma cells, and to provide new ways for glioma immune therapy. Methods: DCs originated from the HLA-A2 +PBMCs were infected with recombinant adenovirus containing EPHA2 full length cDNA, and the DC vaccine modified by EPHA2 gene was prepared. The expression of EPHA on DCs was detected by Western blotting and FACS. HLA-A2 +PBMCs were stimulated by the DC vaccine in vitro. The specificity CTL activity induced by rAd- EPHA2 -DCs and the cytotoxicity on HLA-A2 +U251 glima cells were detected by enzyme-linked immunospot assay (ELISPOT) and standard 51 Cr release experiment. Results: The DC vaccine modified by EPHA2 gene was successfully prepared, and EPHA2 protein was effectively expressed. Compared to DCs infected with rAd-LacZ and PBS groups, DCs infected with rAd-EPHA2 stimulated CTL activity effectively (\[187±21\] vs \[12±4\], \[18±5\]; P<0.01) and the CTLs induced by rAd-EPHA2-DCs produced cytotoxicity effect on U251 cells obviously (\[45.7±6.8\]% vs \[7.1±45\]%, P<0.01), and did not cause the cytotoxicity of its own lymphocytes. Conclusion: The DC vaccine modified by EPHA2 gene can stimulate the specificity CTL response effectively, and can cause cytotoxicity on glioma U251 cells obviously.
Objective : To explore the cytotoxic effect of cyctotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) modified by EPHA2 gene on U251 glioma cells, and to provide new ways for glioma immune therapy. Methods: DCs originated from the HLA-A2 +PBMCs were infected with recombinant adenovirus containing EPHA2 full length cDNA, and the DC vaccine modified by EPHA2 gene was prepared. The expression of EPHA on DCs was detected by Western blotting and FACS. HLA-A2 +PBMCs were stimulated by the DC vaccine in vitro. The specificity CTL activity induced by rAd- EPHA2 -DCs and the cytotoxicity on HLA-A2 +U251 glima cells were detected by enzyme-linked immunospot assay (ELISPOT) and standard 51 Cr release experiment. Results: The DC vaccine modified by EPHA2 gene was successfully prepared, and EPHA2 protein was effectively expressed. Compared to DCs infected with rAd-LacZ and PBS groups, DCs infected with rAd-EPHA2 stimulated CTL activity effectively (\[187±21\] vs \[12±4\], \[18±5\]; P<0.01) and the CTLs induced by rAd-EPHA2-DCs produced cytotoxicity effect on U251 cells obviously (\[45.7±6.8\]% vs \[7.1±45\]%, P<0.01), and did not cause the cytotoxicity of its own lymphocytes. Conclusion: The DC vaccine modified by EPHA2 gene can stimulate the specificity CTL response effectively, and can cause cytotoxicity on glioma U251 cells obviously.
2012, 19(1):40-44.
DOI: 10.3872/j.issn.1007-385X.2012.1.007
Abstract:
Objective : To use G250 monoclonal antibody (G250 mAb) IgG-complexed renal carcinoma cells to acti-vate dendritic cells (DCs), develop the DC vaccine of renal carcinoma and determine its activation for clinical tumor biological therapy. Methods: Prepare apoptotic renal carcinoma cells and induce the G250 mAb-complexed apoptotic renal carcinoma cells (IC-ATC). Immature dendritic cells (iDCs) induced from peripheral blood mononuclear cells of healthy volunteers were cultured and propagated in vitro using rhGM-CSF and rhIL-4. iDCs were loaded with apoptotic renal carcinoma cells (ATC), IC-ATC and G250 mAb. Then mature dendritic cells (mDCs) were induced by TNF-α, while the DCs pulsed with no antigen served as a control group. The immune phenotype of mDCs in different groups was detected by flow cytometry, secretion of IL-12 by DCs was measured by ELISA and the ability of DCs to stimulate lymphocyte proliferation was examined by CCK-8 assay. Results: It was found that when compared with ATC, G250 mAb and the control group, the mDCs pulsed with IC-ATC obriously up-regulated the expressions of CD83, CD80,CD86, and HLA-DR (\[4204±342\]% vs \[28.34±1.16\]%, \[33.77±1.61\]%, \[2652±2.14\]%, P<0.05；\[38.17±2.55\]% vs \[23.79±241\]%, \[31.94±3.29\]%, \[24.32±3.23\]%, P<0.05；\[79.39±1.44\]% vs \[69.06±2.01\]%, \[7449±135\]%, \[66.71±3.83\]%， P<0.05；\[35.52±2.72\]% vs \[26.90±2.82\]%, \[29.45±1.58\]%, \[27.42±211\]%, P<0.05), and secreted higher quantity of IL-12(25.04 vs 5.27, 13.32, 7.53, P＜0.05). Moreover, mDCs loaded by IC-ATC induced multiplication of lymphocytes was more effective (4.02 vs 1.73, 1.22 vs 1.41, P<0.05）. Conclusion: IC-ATC can promote DCs mature effectively, and DCs pulsed with IC-ATC can induce the proliferation and activation significantly.
Objective : To use G250 monoclonal antibody (G250 mAb) IgG-complexed renal carcinoma cells to acti-vate dendritic cells (DCs), develop the DC vaccine of renal carcinoma and determine its activation for clinical tumor biological therapy. Methods: Prepare apoptotic renal carcinoma cells and induce the G250 mAb-complexed apoptotic renal carcinoma cells (IC-ATC). Immature dendritic cells (iDCs) induced from peripheral blood mononuclear cells of healthy volunteers were cultured and propagated in vitro using rhGM-CSF and rhIL-4. iDCs were loaded with apoptotic renal carcinoma cells (ATC), IC-ATC and G250 mAb. Then mature dendritic cells (mDCs) were induced by TNF-α, while the DCs pulsed with no antigen served as a control group. The immune phenotype of mDCs in different groups was detected by flow cytometry, secretion of IL-12 by DCs was measured by ELISA and the ability of DCs to stimulate lymphocyte proliferation was examined by CCK-8 assay. Results: It was found that when compared with ATC, G250 mAb and the control group, the mDCs pulsed with IC-ATC obriously up-regulated the expressions of CD83, CD80,CD86, and HLA-DR (\[4204±342\]% vs \[28.34±1.16\]%, \[33.77±1.61\]%, \[2652±2.14\]%, P<0.05；\[38.17±2.55\]% vs \[23.79±241\]%, \[31.94±3.29\]%, \[24.32±3.23\]%, P<0.05；\[79.39±1.44\]% vs \[69.06±2.01\]%, \[7449±135\]%, \[66.71±3.83\]%， P<0.05；\[35.52±2.72\]% vs \[26.90±2.82\]%, \[29.45±1.58\]%, \[27.42±211\]%, P<0.05), and secreted higher quantity of IL-12(25.04 vs 5.27, 13.32, 7.53, P＜0.05). Moreover, mDCs loaded by IC-ATC induced multiplication of lymphocytes was more effective (4.02 vs 1.73, 1.22 vs 1.41, P<0.05）. Conclusion: IC-ATC can promote DCs mature effectively, and DCs pulsed with IC-ATC can induce the proliferation and activation significantly.
TANG Zhi-wei
,
LI Jie-yu
,
PENG Feng
,
ZHAO Shen
,
ZHOU Zhi-feng
,
CHEN Ming-shui
,
YE Yun-bin
,
CHEN Qiang
2012, 19(1):45-51.
DOI: 10.3872/j.issn.1007-385X.2012.1.008
Abstract:
Objective : To analyze the effect of irradiated haploidentical lymphocyte infusion (HLI) following different doses of cyclophosphamide （CTX） on hepatocellular carcinoma Hepal-6 cells. Methods: Female (BABL/c×C57BL)→F1(CB6F1 H-2 b/d ) mice were subcutaneously inoculated with Hepa1-6 cells to develop a solid tumor model that was taken as recipients. The irradiated haploidentical lymphocytes were from the female (BALB/c×C3H)→F1(CC3HF1 H-2 d/k ).All the mice were divided to five groups: group PBS, group CTX 80 mg/kg+SP/irr(spleen lymphocyte/5Gy irradiated), group CTX 200 mg/kg+SP/irr, group CTX 300 mg/kg+SP/irr and group SP/irr. The tumor size was measured every 2-3 days. The chimerism was measured after HLI, and the graft-versus host disease (GVHD) was assessed. Results: The tumor was significantly suppressed in group CTX 80 mg/kg+SP/irr and group CTX 200 mg/kg+SP/irr compared with group PBS (\[1.25±0.24\] cm 3, \[1.38±0.31\] cm 3 vs \[2.03±0.24\] cm 3, P<0.01), so was the survival time （48 d \[39 d, 55 d\], 40 d \[35 d,48 d\] vs 35 d \[18 d, 39 d\], P<0.05), of which, group CTX 80 mg/kg+SP/irr got significantly different antitumor effect from the group SP/irr (\[1.25±0.24\] cm 3 vs \[1.76±0.40\] cm 3，P<0.05). GVHD was not found in any groups. The chimerism of group 80 mg and 200 mg CTX combining with HLI was lower than group CTX 300 mg combining with HLI and the disappearence time was shorter. Conclusion: Transplantation of irradiated haploidentical lymphocytes following low does of CTX can induce antitumor effect on hepatocellular carcinoma of mice. Higher doses of CTX may not increase antitumor effect.
Objective : To analyze the effect of irradiated haploidentical lymphocyte infusion (HLI) following different doses of cyclophosphamide （CTX） on hepatocellular carcinoma Hepal-6 cells. Methods: Female (BABL/c×C57BL)→F1(CB6F1 H-2 b/d ) mice were subcutaneously inoculated with Hepa1-6 cells to develop a solid tumor model that was taken as recipients. The irradiated haploidentical lymphocytes were from the female (BALB/c×C3H)→F1(CC3HF1 H-2 d/k ).All the mice were divided to five groups: group PBS, group CTX 80 mg/kg+SP/irr(spleen lymphocyte/5Gy irradiated), group CTX 200 mg/kg+SP/irr, group CTX 300 mg/kg+SP/irr and group SP/irr. The tumor size was measured every 2-3 days. The chimerism was measured after HLI, and the graft-versus host disease (GVHD) was assessed. Results: The tumor was significantly suppressed in group CTX 80 mg/kg+SP/irr and group CTX 200 mg/kg+SP/irr compared with group PBS (\[1.25±0.24\] cm 3, \[1.38±0.31\] cm 3 vs \[2.03±0.24\] cm 3, P<0.01), so was the survival time （48 d \[39 d, 55 d\], 40 d \[35 d,48 d\] vs 35 d \[18 d, 39 d\], P<0.05), of which, group CTX 80 mg/kg+SP/irr got significantly different antitumor effect from the group SP/irr (\[1.25±0.24\] cm 3 vs \[1.76±0.40\] cm 3，P<0.05). GVHD was not found in any groups. The chimerism of group 80 mg and 200 mg CTX combining with HLI was lower than group CTX 300 mg combining with HLI and the disappearence time was shorter. Conclusion: Transplantation of irradiated haploidentical lymphocytes following low does of CTX can induce antitumor effect on hepatocellular carcinoma of mice. Higher doses of CTX may not increase antitumor effect.
ZHAO Zhi-ning
,
ZHOU Qiang
,
BAI Jiu-xu
,
YAN Bo
,
QIN Wei-wei
,
WANG Tao
,
JIA Lin-tao
,
YANG An-gang
2012, 19(1):52-55.
DOI: 10.3872/j.issn.1007-385X.2012.1.009
Abstract:
Objective : To observe the effect of Trichostatin A (TSA) on the apoptosis of endometrial cancer Ishikawa cells and to study its relationship with Krupell-like-factor 4 (KLF4) in this course. Methods: Ishikawa cells were cultured with different concentrations of TSA 0, 50, 100, 200, 300, 500 ng/ml for 24 h or 100 ng/ml TSA for 0, 4, 8, 12, 24 and 48 h. FACS and qRT-PCR were used to detect apoptosis and KLF4 mRNA level, respectively. Results: The apoptosis rate was increased compared to the control in the Ishikawa cells treated with 100 ng/ml TSA for 24 h （\[30.6±45\]%vs \[7.53±0.93\]%, P<0.05). The mRNA levels of KLF4 were up-regulated after Ishikawa cells were stimulated with different concentrations of TSA for 24 h or with 100 ng/ml TSA for 4, 8, 12, 24, 48 h (P<0.05). Those effects were in a dose-dependent or time-dependent manner. The apoptosis rate was increased compared to the control in the Ishikawa cells over-expressed KLF4 （\[27.3±27\]% vs \[4.53±1.75\]%, P<0.05). Conclusion: TSA induces apoptosis of Ishikawa cells by up-regulating the expression of KLF4.
Objective : To observe the effect of Trichostatin A (TSA) on the apoptosis of endometrial cancer Ishikawa cells and to study its relationship with Krupell-like-factor 4 (KLF4) in this course. Methods: Ishikawa cells were cultured with different concentrations of TSA 0, 50, 100, 200, 300, 500 ng/ml for 24 h or 100 ng/ml TSA for 0, 4, 8, 12, 24 and 48 h. FACS and qRT-PCR were used to detect apoptosis and KLF4 mRNA level, respectively. Results: The apoptosis rate was increased compared to the control in the Ishikawa cells treated with 100 ng/ml TSA for 24 h （\[30.6±45\]%vs \[7.53±0.93\]%, P<0.05). The mRNA levels of KLF4 were up-regulated after Ishikawa cells were stimulated with different concentrations of TSA for 24 h or with 100 ng/ml TSA for 4, 8, 12, 24, 48 h (P<0.05). Those effects were in a dose-dependent or time-dependent manner. The apoptosis rate was increased compared to the control in the Ishikawa cells over-expressed KLF4 （\[27.3±27\]% vs \[4.53±1.75\]%, P<0.05). Conclusion: TSA induces apoptosis of Ishikawa cells by up-regulating the expression of KLF4.
10 Construction of adenovector expressing small hairpin RNA targeting Bcl-XL and its anti-tumor effect
2012, 19(1):61-65.
DOI: 10.3872/j.issn.1007-385X.2012.1.011
Abstract:
Objective : To construct the adenovector expressing small hairpin RNA targeting Bcl-XL (Ad/Bcl-XL shRNA ), and evaluate its anti-tumor effect. Methods: Firstly, Ad/Bcl-XL shRNA was constructed and purified. Then the protein level of Bcl-XL and survival of colon cancer cells after the treatment of Ad/Bcl-XL shRNA were determined by Western blotting and MTT assay, respectively. Furthermore, the activation of apoptotic signaling was also detected by Western blotting assay. Finally, the anticancer effect of Ad/Bcl-XL shRNA in vivo was confirmed in the subcutaneous tumor model derived from DLD1 cells in nude mice. Results: Ad/Bcl-XL shRNA was constructed and purified successfully. It obviously down-regulated the Bcl-XL protein and significantly inhibited the growth of DLD1 cells (1 000 MOI and 2 000 MOI Ad/Bcl-XL shRNA group was (MOI=1 000: \[60.6±4.8\]% vs \[37.3±6.9\]%， 100%; MOI=2 000: \[99.0±26\]% vs \[ 99.0± 2.1\]%, 100% P<0.01), but had no obvious toxicity on normal human fibroblasts. Western blotting results demonstrated that the apoptotic signal molecules including caspase-9， caspase-3, and PARP were obviously activated after the treatment with Ad/Bcl-XL shRNA. In vivo, it also dramatically suppressed the growth of subcutaneous tumors derived from DLD1 cells in nude mice (eg.29th day Ad/Bcl-XL shRNA group was \[250.1±185.7\] vs Ad/GFP \[880.0±286.1\]，PBS \[ 9110± 389.1\] mm 3，P<0.01). Conclusion: Ad/Bcl-XL shRNA can down-regulate the expression of Bcl-XL and inhibit the growth of colon cancer cells in vivo and in vitro, suggesting that it may be a new strategy to treat the colon carcinoma.
Objective : To construct the adenovector expressing small hairpin RNA targeting Bcl-XL (Ad/Bcl-XL shRNA ), and evaluate its anti-tumor effect. Methods: Firstly, Ad/Bcl-XL shRNA was constructed and purified. Then the protein level of Bcl-XL and survival of colon cancer cells after the treatment of Ad/Bcl-XL shRNA were determined by Western blotting and MTT assay, respectively. Furthermore, the activation of apoptotic signaling was also detected by Western blotting assay. Finally, the anticancer effect of Ad/Bcl-XL shRNA in vivo was confirmed in the subcutaneous tumor model derived from DLD1 cells in nude mice. Results: Ad/Bcl-XL shRNA was constructed and purified successfully. It obviously down-regulated the Bcl-XL protein and significantly inhibited the growth of DLD1 cells (1 000 MOI and 2 000 MOI Ad/Bcl-XL shRNA group was (MOI=1 000: \[60.6±4.8\]% vs \[37.3±6.9\]%， 100%; MOI=2 000: \[99.0±26\]% vs \[ 99.0± 2.1\]%, 100% P<0.01), but had no obvious toxicity on normal human fibroblasts. Western blotting results demonstrated that the apoptotic signal molecules including caspase-9， caspase-3, and PARP were obviously activated after the treatment with Ad/Bcl-XL shRNA. In vivo, it also dramatically suppressed the growth of subcutaneous tumors derived from DLD1 cells in nude mice (eg.29th day Ad/Bcl-XL shRNA group was \[250.1±185.7\] vs Ad/GFP \[880.0±286.1\]，PBS \[ 9110± 389.1\] mm 3，P<0.01). Conclusion: Ad/Bcl-XL shRNA can down-regulate the expression of Bcl-XL and inhibit the growth of colon cancer cells in vivo and in vitro, suggesting that it may be a new strategy to treat the colon carcinoma.
2012, 19(1):66-71.
DOI: 10.3872/j.issn.1007-385X.2012.1.012
Abstract:
Objective : To construct lentiviral expressed vector of siRNA targeting CIAPIN1 and establish breast cancer cells with a stable expression of siRNA- CIAPIN1 , and to investigate the effect of CIAPIN1 on breast cancer cells multi-drug resistance. Methods: The expressed vectors of recombined plasmid of siRNA targeting CIAPIN1 were designed and synthesized. Select the most efficient interfering sequence, spackage, and produce a lentiviral vector with it. Stably transfect CIAPIN1 -RNAi into MCF-7/ADM cells. Detect IC 50 value of different drugs in MCF-7/ADM cells before and after CIAPIN1 interference by MTT. Results: The expressed vectors of recombined plasmid of siRNA targeting CIAPIN1 were successfully synthesized and the most efficient interfering sequence was CIAPIN1 -siRNA1. Stable transfection of CIAPIN1 -RNAi into MCF-7/ADM cells by a lentiviral vector suppressed the expression of CIAPIN1 in MCF-7/ADM cells more than 88%. After RNA interference, IC 50 value of MCF-7/ADM cells to anticancer drugs (paclitaxel, doxorubicin and gemcitabine) significantly decreased from (7.12±0.31), (11.21±1.79), (49.72±4.52) to (1.13±0.06), (4.51±020), (1830±127) μg/ml respectively, suggesting a significant decrease in the drug resisstance of the cells. Conclusion: Lentiviral expressed vector of CIAPIN1 -siRNA can efficiently interfere the expression of CIAPIN1 in MCF-7/ADM cells. The study also confirmed the regulation effect of CIAPIN1 on breast cancer cell multi-drug resistance (MDR).
Objective : To construct lentiviral expressed vector of siRNA targeting CIAPIN1 and establish breast cancer cells with a stable expression of siRNA- CIAPIN1 , and to investigate the effect of CIAPIN1 on breast cancer cells multi-drug resistance. Methods: The expressed vectors of recombined plasmid of siRNA targeting CIAPIN1 were designed and synthesized. Select the most efficient interfering sequence, spackage, and produce a lentiviral vector with it. Stably transfect CIAPIN1 -RNAi into MCF-7/ADM cells. Detect IC 50 value of different drugs in MCF-7/ADM cells before and after CIAPIN1 interference by MTT. Results: The expressed vectors of recombined plasmid of siRNA targeting CIAPIN1 were successfully synthesized and the most efficient interfering sequence was CIAPIN1 -siRNA1. Stable transfection of CIAPIN1 -RNAi into MCF-7/ADM cells by a lentiviral vector suppressed the expression of CIAPIN1 in MCF-7/ADM cells more than 88%. After RNA interference, IC 50 value of MCF-7/ADM cells to anticancer drugs (paclitaxel, doxorubicin and gemcitabine) significantly decreased from (7.12±0.31), (11.21±1.79), (49.72±4.52) to (1.13±0.06), (4.51±020), (1830±127) μg/ml respectively, suggesting a significant decrease in the drug resisstance of the cells. Conclusion: Lentiviral expressed vector of CIAPIN1 -siRNA can efficiently interfere the expression of CIAPIN1 in MCF-7/ADM cells. The study also confirmed the regulation effect of CIAPIN1 on breast cancer cell multi-drug resistance (MDR).
2012, 19(1):72-76.
DOI: 10.3872/j.issn.1007-385X.2012.1.013
Abstract:
Objective : To investigate the treatment effect of recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA targeting hTERT gene on human colorectal cancer SW480 cell xenograft in nude mice. Methods: Human colorectal cancer SW480 cells were subcutaneously implanted under the skin of the right armpit to establish nude mice model of colorectal cancer, after the tumors grew to a definite size. The mice were randomly divided into three groups: normal saline (NS group), pGPU6/GFP/Neo-NC-shRNA group（NC-shRNA group）and pGPU6/GFP/Neo-hTERT-shRNA group（hTERT-shRNA group）. After each group was treated for 6 consecutive times, the growth status of the tumor was observed, tumor volume was measured, tumor growth curve was drawn，tumor tissue morphology was observed with H-E staining, the expression of hTERT protein in the tumors was detected by immunohistochemistry, cell apoptosis was inspected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling(TUNEL), and the expression of hTERT mRNA was checked by RT-PCR. Results: The growth of tumor volume became slower in hTERT-shRNA group than did that in NS group and NC-shRNA group. Compared with NS group and NC-shRNA group, the tumor cell morphology changed obviously and the number of apoptotic cells increased significantly in the transplanted tumor tissues in hTERT-shRNA group（\[363±505\]% vs \[5.25± 1.06\]%, \[6.95±1.07\]%，P<0.01). Compared with NS group and NC-shRNA group, the expression of hTERT protein was significantly inhibited in hTERT-shRNA group（\[171.42±30.94\] vs \[14689±21.43\], \[13735±25.49\], P<0.01）. Compared with NS group and NC-shRNA group, the expression of hTERT mRNA was significantly inhibited in hTERT-shRNA group （0.39±0.09 vs 0.81±0.34, 0.75±0.21, P<0.05）. Conclusion: Recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA promotes apoptosis of implanted human colorectal cancer by down-regulating the expression of hTERT mRNA and hTERT protein in tumor tissues, thus inhibiting the growth.
Objective : To investigate the treatment effect of recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA targeting hTERT gene on human colorectal cancer SW480 cell xenograft in nude mice. Methods: Human colorectal cancer SW480 cells were subcutaneously implanted under the skin of the right armpit to establish nude mice model of colorectal cancer, after the tumors grew to a definite size. The mice were randomly divided into three groups: normal saline (NS group), pGPU6/GFP/Neo-NC-shRNA group（NC-shRNA group）and pGPU6/GFP/Neo-hTERT-shRNA group（hTERT-shRNA group）. After each group was treated for 6 consecutive times, the growth status of the tumor was observed, tumor volume was measured, tumor growth curve was drawn，tumor tissue morphology was observed with H-E staining, the expression of hTERT protein in the tumors was detected by immunohistochemistry, cell apoptosis was inspected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling(TUNEL), and the expression of hTERT mRNA was checked by RT-PCR. Results: The growth of tumor volume became slower in hTERT-shRNA group than did that in NS group and NC-shRNA group. Compared with NS group and NC-shRNA group, the tumor cell morphology changed obviously and the number of apoptotic cells increased significantly in the transplanted tumor tissues in hTERT-shRNA group（\[363±505\]% vs \[5.25± 1.06\]%, \[6.95±1.07\]%，P<0.01). Compared with NS group and NC-shRNA group, the expression of hTERT protein was significantly inhibited in hTERT-shRNA group（\[171.42±30.94\] vs \[14689±21.43\], \[13735±25.49\], P<0.01）. Compared with NS group and NC-shRNA group, the expression of hTERT mRNA was significantly inhibited in hTERT-shRNA group （0.39±0.09 vs 0.81±0.34, 0.75±0.21, P<0.05）. Conclusion: Recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA promotes apoptosis of implanted human colorectal cancer by down-regulating the expression of hTERT mRNA and hTERT protein in tumor tissues, thus inhibiting the growth.
13 Expressions of KISS1 and osteopontin in epithelial ovarian cancer and their clinical significance
2012, 19(1):77-80.
DOI: 10.3872/j.issn.1007-385X.2012.1.014
Abstract:
Objective : To investigate the expression and clinical significance of KiSS-1 metastasis-suppressor (KISS1) and osteopontin (OPN) in epithelial ovarian cancer (EOC). Methods: From March 2009 to October 2010, epithelial ovarian tumors of 67 patients operated in Gynecology Department of the Fourth Hospital of Hebei Medical University were selected. The expressions of KISS1 and OPN in EOC were detected by immunohistochemistry (IHC). Results: The frequency of KISS1 positive expression was 39.53% (17/43) in EOC tissues, which was significantly lower than that in ovarian benign tumor tissues (75.00%, 18/24), with a significant difference between two groups (χ 2=7.765, P=0005). The expression of KISS1 protein in the lymph node metastasis group was significantly lower than that in the without lymph node metastasis group (250% \[7/28\] vs 66.7% \[10/15\]; χ 2=7.094, P=0.008). In the clinical stage group, the expression of KISS1 protein was significantly higher in Ⅰ+Ⅱ stage than in Ⅲ+Ⅳ stage (61.1% \[11/18\] vs 24.0% \[6/25\]; χ 2=6.029, P=0.014). The frequency of OPN positive expression was 74.42% (32/43) in EOC tissues, which was significantly higher than that in the ovarian benign tumor tissue group (37.50%, 11/24) with a significant difference between two groups（74.4% \[32/43\] vs 37.5%\[11/24\];χ 2=5.475, P=0.019）. The expression of OPN in the lymph node metastasis group was significantly higher than that in the without lymph node metastasis group (893% \[25/28\] vs 46.7% \[7/15\]; χ2=7.251, P=0.007). In the clinical stage group, the expression of OPN was significantly lower in Ⅰ+Ⅱ stage than in Ⅲ+Ⅳ stage (50.0% \[9/18\] vs 92.0 \[23/25\]; χ 2=7.616, P=0.006). The protein expressions of KISS1 and OPN were not correlated with pathologic classification and patient’s age of EOC （ P＞005 ）, and there was a negative correlation between KISS1 and OPN protein expressions (r=-0.507, P=0001). Conclusion: The protein expressions of OPN and KISS1 may participate in the carcinogenesis, invasion and metastasis of EOC, which may contribute to prognosis marker of EOC.
Objective : To investigate the expression and clinical significance of KiSS-1 metastasis-suppressor (KISS1) and osteopontin (OPN) in epithelial ovarian cancer (EOC). Methods: From March 2009 to October 2010, epithelial ovarian tumors of 67 patients operated in Gynecology Department of the Fourth Hospital of Hebei Medical University were selected. The expressions of KISS1 and OPN in EOC were detected by immunohistochemistry (IHC). Results: The frequency of KISS1 positive expression was 39.53% (17/43) in EOC tissues, which was significantly lower than that in ovarian benign tumor tissues (75.00%, 18/24), with a significant difference between two groups (χ 2=7.765, P=0005). The expression of KISS1 protein in the lymph node metastasis group was significantly lower than that in the without lymph node metastasis group (250% \[7/28\] vs 66.7% \[10/15\]; χ 2=7.094, P=0.008). In the clinical stage group, the expression of KISS1 protein was significantly higher in Ⅰ+Ⅱ stage than in Ⅲ+Ⅳ stage (61.1% \[11/18\] vs 24.0% \[6/25\]; χ 2=6.029, P=0.014). The frequency of OPN positive expression was 74.42% (32/43) in EOC tissues, which was significantly higher than that in the ovarian benign tumor tissue group (37.50%, 11/24) with a significant difference between two groups（74.4% \[32/43\] vs 37.5%\[11/24\];χ 2=5.475, P=0.019）. The expression of OPN in the lymph node metastasis group was significantly higher than that in the without lymph node metastasis group (893% \[25/28\] vs 46.7% \[7/15\]; χ2=7.251, P=0.007). In the clinical stage group, the expression of OPN was significantly lower in Ⅰ+Ⅱ stage than in Ⅲ+Ⅳ stage (50.0% \[9/18\] vs 92.0 \[23/25\]; χ 2=7.616, P=0.006). The protein expressions of KISS1 and OPN were not correlated with pathologic classification and patient’s age of EOC （ P＞005 ）, and there was a negative correlation between KISS1 and OPN protein expressions (r=-0.507, P=0001). Conclusion: The protein expressions of OPN and KISS1 may participate in the carcinogenesis, invasion and metastasis of EOC, which may contribute to prognosis marker of EOC.
MA Xin-fu
,
ZHANG Cheng-wu
,
WEN Ying
,
ZHAO Jiu-da
,
YAN Su
,
CAI Bao-jia
,
MIAO Wei
,
WANG Xiao-long
2012, 19(1):81-86.
DOI: 10.3872/j.issn.1007-385X.2012.1.015
Abstract:
Objective : To quantitatively assess the efficacy of interferon (IFN) in the adjuvant treatment for patients with hepatocellular carcinoma (HCC) using Meta analysis. Methods: Randomized controlled trials (RCTs) of adjuvant treatment with interferon for patients with HCC were searched in Embase, Medline, Cochrane Library, Chinese biomedicine literature database, Chinese Scientific Journals full-text database, and Chinese Journal full-text databases. Two reviewers independently assessed the quality of included studies and extracted data. Meta analysis was carried out using Review Manager (version 5.0) provided by Cochrane Collaboration. Results: Eight RCTs totaling 836 patients were included. Meta analysis showed that the adjuvant treatment with IFN significantly reduced the recurrence rate after curative treatment of HCC, with a pooled risk ratio of 0.86 (95 percent confidence interval 0.77 to 0.96); the effect on reduction in mortality rate was still significant with a pooled risk ratio of 0.64 (95 percent confidence interval 0.54 to 0.76). Conclusion: IFN has a beneficial effect on both mortality rate and tumour recurrence, and the results still need to be confirmed by RCTs of high quality and large sample size.
Objective : To quantitatively assess the efficacy of interferon (IFN) in the adjuvant treatment for patients with hepatocellular carcinoma (HCC) using Meta analysis. Methods: Randomized controlled trials (RCTs) of adjuvant treatment with interferon for patients with HCC were searched in Embase, Medline, Cochrane Library, Chinese biomedicine literature database, Chinese Scientific Journals full-text database, and Chinese Journal full-text databases. Two reviewers independently assessed the quality of included studies and extracted data. Meta analysis was carried out using Review Manager (version 5.0) provided by Cochrane Collaboration. Results: Eight RCTs totaling 836 patients were included. Meta analysis showed that the adjuvant treatment with IFN significantly reduced the recurrence rate after curative treatment of HCC, with a pooled risk ratio of 0.86 (95 percent confidence interval 0.77 to 0.96); the effect on reduction in mortality rate was still significant with a pooled risk ratio of 0.64 (95 percent confidence interval 0.54 to 0.76). Conclusion: IFN has a beneficial effect on both mortality rate and tumour recurrence, and the results still need to be confirmed by RCTs of high quality and large sample size.
2012, 19(1):87-92.
DOI: 10.3872/j.issn.1007-385X.2012.1.016
Abstract:
炎症反应在清除病原体、创伤愈合和抗肿瘤免疫中发挥重要作用，被肿瘤细胞劫持的炎性细胞和细胞因子也是肿瘤微环境重要的参与者；许多肿瘤起源于感染和慢性炎症，肿瘤微环境中炎症细胞和炎症介质的蓄积具有促进恶性细胞增殖和存活、促进血管生成和肿瘤转移，以及逆转获得性免疫反应的作用，也改变了肿瘤细胞对激素和化疗药物的敏感性。炎症反应在肿瘤的发生和消除中发挥的作用比较复杂。溶瘤病毒治疗肿瘤，是充分利用溶瘤病毒选择性感染和杀伤肿瘤细胞的特性。在肿瘤微环境中，溶瘤病毒所诱导的针对肿瘤细胞和病毒的天然免疫反应具有双重效应：既能引起肿瘤细胞的损伤，促进溶瘤病毒抗肿瘤的疗效，也能识别、清除隐藏于肿瘤组织内部的溶瘤病毒，降低溶瘤病毒的效应。
炎症反应在清除病原体、创伤愈合和抗肿瘤免疫中发挥重要作用，被肿瘤细胞劫持的炎性细胞和细胞因子也是肿瘤微环境重要的参与者；许多肿瘤起源于感染和慢性炎症，肿瘤微环境中炎症细胞和炎症介质的蓄积具有促进恶性细胞增殖和存活、促进血管生成和肿瘤转移，以及逆转获得性免疫反应的作用，也改变了肿瘤细胞对激素和化疗药物的敏感性。炎症反应在肿瘤的发生和消除中发挥的作用比较复杂。溶瘤病毒治疗肿瘤，是充分利用溶瘤病毒选择性感染和杀伤肿瘤细胞的特性。在肿瘤微环境中，溶瘤病毒所诱导的针对肿瘤细胞和病毒的天然免疫反应具有双重效应：既能引起肿瘤细胞的损伤，促进溶瘤病毒抗肿瘤的疗效，也能识别、清除隐藏于肿瘤组织内部的溶瘤病毒，降低溶瘤病毒的效应。
2012, 19(1):93-97.
DOI: 10.3872/j.issn.1007-385X.2012.1.017
Abstract:
钙网蛋白（calreticulin,CRT）为内质网滞留蛋白，属于热激蛋白超家族成员之一，在生物体中参与广泛的生物学作用，其中参与凋亡细胞清除、抗原提呈和诱导特异性免疫应答等被广泛地应用于肿瘤的免疫治疗中。钙网基因疫苗诱导针对肿瘤特异性抗原的免疫应答、细胞膜表面高表达钙网蛋白促进肿瘤细胞被吞噬、细胞内高表达钙网蛋白促进细胞凋亡等已成为抗肿瘤免疫治疗的研究热点。
钙网蛋白（calreticulin,CRT）为内质网滞留蛋白，属于热激蛋白超家族成员之一，在生物体中参与广泛的生物学作用，其中参与凋亡细胞清除、抗原提呈和诱导特异性免疫应答等被广泛地应用于肿瘤的免疫治疗中。钙网基因疫苗诱导针对肿瘤特异性抗原的免疫应答、细胞膜表面高表达钙网蛋白促进肿瘤细胞被吞噬、细胞内高表达钙网蛋白促进细胞凋亡等已成为抗肿瘤免疫治疗的研究热点。
2012, 19(1):98-102.
DOI: 10.3872/j.issn.1007-385X.2012.1.018
Abstract:
肿瘤的早发现、早治疗及个性化诊疗方案是肿瘤治疗的发展方向，新兴的代谢组学(metabolomics)开辟了辅助肿瘤早期诊断、疗效评价及预后判断的新思路。代谢组学是一种将图像识别方法和生物信息学结合起来的分析技术，用于检测体液或组织中的代谢产物并分析其变化规律。肿瘤的发生、发展伴随着机体代谢产物的变化，代谢组学能对肿瘤引起的机体代谢变化作出整体评价，筛选出有价值的生物标志物，用于肿瘤的早期诊治。本文就代谢组学的概念和方法学，代谢组学在肿瘤早期诊断、靶向治疗及其疗效评价、患者预后评估中的作用做一综述。
肿瘤的早发现、早治疗及个性化诊疗方案是肿瘤治疗的发展方向，新兴的代谢组学(metabolomics)开辟了辅助肿瘤早期诊断、疗效评价及预后判断的新思路。代谢组学是一种将图像识别方法和生物信息学结合起来的分析技术，用于检测体液或组织中的代谢产物并分析其变化规律。肿瘤的发生、发展伴随着机体代谢产物的变化，代谢组学能对肿瘤引起的机体代谢变化作出整体评价，筛选出有价值的生物标志物，用于肿瘤的早期诊治。本文就代谢组学的概念和方法学，代谢组学在肿瘤早期诊断、靶向治疗及其疗效评价、患者预后评估中的作用做一综述。
2012, 19(1):103-106.
DOI: 10.3872/j.issn.1007-385X.2012.1.019
Abstract:
肿瘤细胞及其周围免疫细胞分泌的炎性因子可激活炎症相关信号通路，诱导肿瘤免疫抑制微环境的形成，参与结直肠癌的发生，进展和转移的过程。与结直肠癌存在密切联系的炎性因子主要有TNF-α、IL-6、IL-8、TGF-β、PGE2等。TNF-α激活炎症信号转录因子NF-κB和AP-1，进而启动血管形成因子IL-8，促进肿瘤血管生成和肿瘤播散。结肠癌细胞分泌IL-6，激活下游Stat3，刺激自身癌细胞增殖和拮抗凋亡；IL-6还可以通过增强T细胞介导的免疫炎症反应，促进结直肠癌发展。IL-8诱导肿瘤血管增生，介导炎症反应，促进肿瘤侵袭和转移。TGF-β一方面抑制结直肠癌生长，促进肿瘤细胞凋亡；另一方面TGF-β信号通路失活则有助于肿瘤生长，同时TGF-β可以刺激肿瘤血管形成，促进肿瘤转移。PGE2全方位地参与结肠癌发生、发展进程，诱导肿瘤生长微环境形成，促进结直肠癌发展和扩散。
肿瘤细胞及其周围免疫细胞分泌的炎性因子可激活炎症相关信号通路，诱导肿瘤免疫抑制微环境的形成，参与结直肠癌的发生，进展和转移的过程。与结直肠癌存在密切联系的炎性因子主要有TNF-α、IL-6、IL-8、TGF-β、PGE2等。TNF-α激活炎症信号转录因子NF-κB和AP-1，进而启动血管形成因子IL-8，促进肿瘤血管生成和肿瘤播散。结肠癌细胞分泌IL-6，激活下游Stat3，刺激自身癌细胞增殖和拮抗凋亡；IL-6还可以通过增强T细胞介导的免疫炎症反应，促进结直肠癌发展。IL-8诱导肿瘤血管增生，介导炎症反应，促进肿瘤侵袭和转移。TGF-β一方面抑制结直肠癌生长，促进肿瘤细胞凋亡；另一方面TGF-β信号通路失活则有助于肿瘤生长，同时TGF-β可以刺激肿瘤血管形成，促进肿瘤转移。PGE2全方位地参与结肠癌发生、发展进程，诱导肿瘤生长微环境形成，促进结直肠癌发展和扩散。
2012, 19(1):107-109.
DOI: 10.3872/j.issn.1007-385X.2012.1.020
Abstract:
胶质瘤干细胞（glioma stem cells，GSC）是近年来在胶质瘤组织中发现的肿瘤干细胞。GSC具有自我更新和分化的能力，可以通过不断分化产生新的肿瘤；对GSC标志物的鉴定有助于胶质瘤恶性程度的诊断。GSC的起源目前仍不明确，成熟的神经胶质细胞、限制性神经祖细胞以及神经干细胞均可能为GSC的前体。推测如Wnt、Notch、SHH、BMI1、PTEN等信号通路活跃在GSC中，一些新的治疗手段通过作用于GSC的信号转导通路可靶向治疗胶质瘤。深入研究GSC的起源、标志物以及相关信号转导通路，可为胶质瘤治疗提供新的策略。
胶质瘤干细胞（glioma stem cells，GSC）是近年来在胶质瘤组织中发现的肿瘤干细胞。GSC具有自我更新和分化的能力，可以通过不断分化产生新的肿瘤；对GSC标志物的鉴定有助于胶质瘤恶性程度的诊断。GSC的起源目前仍不明确，成熟的神经胶质细胞、限制性神经祖细胞以及神经干细胞均可能为GSC的前体。推测如Wnt、Notch、SHH、BMI1、PTEN等信号通路活跃在GSC中，一些新的治疗手段通过作用于GSC的信号转导通路可靶向治疗胶质瘤。深入研究GSC的起源、标志物以及相关信号转导通路，可为胶质瘤治疗提供新的策略。
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- 《中国肿瘤生物治疗杂志》
- 1994年创刊
- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
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- 刊号：ISSN 1007-385X
- CN 31-1725/R
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.