Mobile website
Volume 17,Issue 1,2010 Table of Contents
2010, 17(1):1-6.
DOI: 10.3872/j.issn.1007-385X.2010.1.001
Abstract:
There are multiple types of inhibitory immune cells in tumor. Among these cells, Treg (regulatory T cell) plays an extremely important role in tumor development and progression. Treg exihibits potent inhibitory effects on effector cells by a variety of mechanisms, which might be the the key factor for tumor immune escape. These mechanisms include inhibiting the effector cell function by inhibitory cytokines, killing effector cells by granzyme and profrin, interfering effector cell metabolism, and affecting Treg differentiation and proliferation by regulating the function of dendrtic cells, etc. The research on Treg has provided new strategies for tumor immunotherapy. Tumor immunotherapies targeting Treg and related immunosuppressive factors, such as deleting Treg nonsepcificlly or sepcificlly controling the numbers and functions of Treg, might have a bright future in clinical application.
There are multiple types of inhibitory immune cells in tumor. Among these cells, Treg (regulatory T cell) plays an extremely important role in tumor development and progression. Treg exihibits potent inhibitory effects on effector cells by a variety of mechanisms, which might be the the key factor for tumor immune escape. These mechanisms include inhibiting the effector cell function by inhibitory cytokines, killing effector cells by granzyme and profrin, interfering effector cell metabolism, and affecting Treg differentiation and proliferation by regulating the function of dendrtic cells, etc. The research on Treg has provided new strategies for tumor immunotherapy. Tumor immunotherapies targeting Treg and related immunosuppressive factors, such as deleting Treg nonsepcificlly or sepcificlly controling the numbers and functions of Treg, might have a bright future in clinical application.
2010, 17(1):7-12.
DOI: 10.3872/j.issn.1007-385X.2010.1.002
Abstract:
Objective:To evaluate the antitumor and side effects of activatedHLA haploidentical peripheral blood tem cells (haploPBSCs) in the treatment of advanced refractory solid tumor patients. Methods:Fortytwo patients with advanced refractory tumor, who were diagnosed in our hospital from Oct. 2004 to Oct. 2007, were enrolled in this study (all patients signed informed consent), including 12 with ovarian cancer, 9 with renal cancer, 8 with lung cancer, 8 with breast cancer, 2 with colon cancer, 2 with gastric cancer, and 1 with melanoma. The donors were healthy direct relatives of the patients; the donors’ haploPBSCs were mobilized, collected, and activated by rhIL2 in vitro. The clinical efficacy and side effects of haploPBSCs therapy were assessed by CT/PETCT scanning, RESIST standard, KPS score, and clinical response rates, etc. Results:All 42 patients received one episode of haploPBSCs treatment. The progressionfree survivals (PFS) were 6 months and the clinical beneficial rate (CR+PR+SD) was 73.8%. The beneficial rate of life quality was 76.2% and the KPS increased by 20 (030) points on average after haploPBSCs treatment. The patients with KIR unmatched in GVH direction had better outcomes than those with KIR matched or KIR unmatched in HVG direction (P<005), and the clinical beneficial rate, PFS and total beneficial rate were 94.1% vs 60.0%, (13.4±1.3) vs (80±09) months, and 89.5% vs 65.2%, respectively (all P<0.05). The donor/recipient relation as the mother/child had a better outcome than that as the father/child (P<0.05). Patients with renal cancer or ovarian cancer had better outcomes than those with other cancers, with clinical beneficial rates being 90.0% and 81.8%, respectively. Conclusion: Activated haploPBSCs therapy can induce nonspecific antitumor effect, and improve the clinical symptom and life quality of advanced tumor patients.
Objective:To evaluate the antitumor and side effects of activatedHLA haploidentical peripheral blood tem cells (haploPBSCs) in the treatment of advanced refractory solid tumor patients. Methods:Fortytwo patients with advanced refractory tumor, who were diagnosed in our hospital from Oct. 2004 to Oct. 2007, were enrolled in this study (all patients signed informed consent), including 12 with ovarian cancer, 9 with renal cancer, 8 with lung cancer, 8 with breast cancer, 2 with colon cancer, 2 with gastric cancer, and 1 with melanoma. The donors were healthy direct relatives of the patients; the donors’ haploPBSCs were mobilized, collected, and activated by rhIL2 in vitro. The clinical efficacy and side effects of haploPBSCs therapy were assessed by CT/PETCT scanning, RESIST standard, KPS score, and clinical response rates, etc. Results:All 42 patients received one episode of haploPBSCs treatment. The progressionfree survivals (PFS) were 6 months and the clinical beneficial rate (CR+PR+SD) was 73.8%. The beneficial rate of life quality was 76.2% and the KPS increased by 20 (030) points on average after haploPBSCs treatment. The patients with KIR unmatched in GVH direction had better outcomes than those with KIR matched or KIR unmatched in HVG direction (P<005), and the clinical beneficial rate, PFS and total beneficial rate were 94.1% vs 60.0%, (13.4±1.3) vs (80±09) months, and 89.5% vs 65.2%, respectively (all P<0.05). The donor/recipient relation as the mother/child had a better outcome than that as the father/child (P<0.05). Patients with renal cancer or ovarian cancer had better outcomes than those with other cancers, with clinical beneficial rates being 90.0% and 81.8%, respectively. Conclusion: Activated haploPBSCs therapy can induce nonspecific antitumor effect, and improve the clinical symptom and life quality of advanced tumor patients.
2010, 17(1):13-18.
DOI: 10.3872/j.issn.1007-385X.2010.1.003
Abstract:
Objective:To investigate the effect of cochinchina momordica seed ethanol extract (CMSEE) on the proliferation of melanoma B16 cells and the underlying mechanism. Methods:MTT and clone formation assay were used to assess the effect of CMSEE on the growth of B16 cells. Morphological changes of B16 cells were observed under phasecontrast microscope and Giemsa staining. Cell cycle and apoptosis rate were examined by flow cytometry (FCM). The effect of CMSEE on melanin production and tyrosinase activity of B16 cells was assessed by colorimetry. The effect of CMSEE on the expression of 〖STBX〗Cmyc, P38, and Tyr〖STBZ〗 genes was examined by RTPCR. Results:CMSEE (10100 mg/L) inhibited the proliferation of B16 cell in a doseand timedependent manner. After treatment with 1040 mg/L CMSEE, B16 cells showed typical differentiation morphology, and melanin production and tyrosinase activity were increased. B16 cells treated with 100 mg/L CMSEE showed apoptotic morphology, decreased melanin production and tyrosinase activity. B16 cell number in G0/G1 phase was significantly increased (P<0.01); Cmyc mRNA expression was downregulated, and P38, Try mRNA expression was upregulated in B16 cells after treatment with 1040 mg/L CMSEE. Conclusion:CMSEE can markedly inhibit the proliferation of melanoma B16 cells, which is related to induction of differentiation and promotion of apoptosis of B16 cells.
Objective:To investigate the effect of cochinchina momordica seed ethanol extract (CMSEE) on the proliferation of melanoma B16 cells and the underlying mechanism. Methods:MTT and clone formation assay were used to assess the effect of CMSEE on the growth of B16 cells. Morphological changes of B16 cells were observed under phasecontrast microscope and Giemsa staining. Cell cycle and apoptosis rate were examined by flow cytometry (FCM). The effect of CMSEE on melanin production and tyrosinase activity of B16 cells was assessed by colorimetry. The effect of CMSEE on the expression of 〖STBX〗Cmyc, P38, and Tyr〖STBZ〗 genes was examined by RTPCR. Results:CMSEE (10100 mg/L) inhibited the proliferation of B16 cell in a doseand timedependent manner. After treatment with 1040 mg/L CMSEE, B16 cells showed typical differentiation morphology, and melanin production and tyrosinase activity were increased. B16 cells treated with 100 mg/L CMSEE showed apoptotic morphology, decreased melanin production and tyrosinase activity. B16 cell number in G0/G1 phase was significantly increased (P<0.01); Cmyc mRNA expression was downregulated, and P38, Try mRNA expression was upregulated in B16 cells after treatment with 1040 mg/L CMSEE. Conclusion:CMSEE can markedly inhibit the proliferation of melanoma B16 cells, which is related to induction of differentiation and promotion of apoptosis of B16 cells.
ZHANG Jun-ling
,
LI Wei-bo
,
XIE Shao-jian
,
LI Dongbin
,
TIAN Qing
,
WANG Yingxia
,
XUE Ping
,
CAI Jian-hui
2010, 17(1):19-24.
DOI: 10.3872/j.issn.1007-385X.2010.1.004
Abstract:
Objective:To investigate the effect of mitoxantrone (MIT) on calreticulin (CRT) expression in B16 cells, and to observe the immune effect of B16membrane antigen vaccine highly expressing CRT on B16 tumorbearing mice. Methods:The expression of CRT on membrane of B16 cells was detected by immunofluorescence after treatment with different concentrations of MIT. B16implanted mouse model was established, and the growth of B16implanted tumors and CRT expression in B16implanted tumor tissues were observed after treatment with different concentrations of MIT. Membrane antigen vaccines from both normal B16 cells and MITtreated B16 cells were prepared, and mice were immunized before B16 cell implantation. The infiltration of immune cells into B16 tumor tissues and the ratios of CD4+ and CD8+ T cells in the spleen of B16 tumorbearing mice were examined by immunohistochemistry and flow cytometry, respectively. Results:Flow cytometry results showed that MIT dosedependently increased CRT expression on B16 cell membrane, with CRT expression in control and high dosage MIT groups being (29.40±3.57)% and (72.20±2.94)% (P<0.05), respectively. MIT also increased CRT expression in B16 tumor tissues, with those in the control and high dosage MIT groups being 3.21±1.37 and 9.17±1.06 (P<0.05), respectively. MIT effectively inhibited the growth of B16 tumors (P<0.05). Compared with normal B16 cell membrane antigen vaccine, the vaccine highly expressing CRT increased the numbers of DCs and T cells in B16 tumors tissues and the ratios of CD4+ and CD8+ T cells in the spleen (P<0.05). Conclusion:MIT can increase CRT expression on membrane of B16 cells. B16membrane antigen vaccine highly expressing CRT can enhance the infiltration of DCs and T cells in melanoma, thus improving the immune effect of B16membrane antigen vaccine.
Objective:To investigate the effect of mitoxantrone (MIT) on calreticulin (CRT) expression in B16 cells, and to observe the immune effect of B16membrane antigen vaccine highly expressing CRT on B16 tumorbearing mice. Methods:The expression of CRT on membrane of B16 cells was detected by immunofluorescence after treatment with different concentrations of MIT. B16implanted mouse model was established, and the growth of B16implanted tumors and CRT expression in B16implanted tumor tissues were observed after treatment with different concentrations of MIT. Membrane antigen vaccines from both normal B16 cells and MITtreated B16 cells were prepared, and mice were immunized before B16 cell implantation. The infiltration of immune cells into B16 tumor tissues and the ratios of CD4+ and CD8+ T cells in the spleen of B16 tumorbearing mice were examined by immunohistochemistry and flow cytometry, respectively. Results:Flow cytometry results showed that MIT dosedependently increased CRT expression on B16 cell membrane, with CRT expression in control and high dosage MIT groups being (29.40±3.57)% and (72.20±2.94)% (P<0.05), respectively. MIT also increased CRT expression in B16 tumor tissues, with those in the control and high dosage MIT groups being 3.21±1.37 and 9.17±1.06 (P<0.05), respectively. MIT effectively inhibited the growth of B16 tumors (P<0.05). Compared with normal B16 cell membrane antigen vaccine, the vaccine highly expressing CRT increased the numbers of DCs and T cells in B16 tumors tissues and the ratios of CD4+ and CD8+ T cells in the spleen (P<0.05). Conclusion:MIT can increase CRT expression on membrane of B16 cells. B16membrane antigen vaccine highly expressing CRT can enhance the infiltration of DCs and T cells in melanoma, thus improving the immune effect of B16membrane antigen vaccine.
2010, 17(1):25-29.
DOI: 10.3872/j.issn.1007-385X.2010.1.005
Abstract:
Objective:To study the therapeutic effect of secondary lymphoid tissue chemokine (SLC) combined with CpG oligodeoxynucleotide (CpGODN) in treatment of implanted mouse melanoma and the possible mechanism. Methods:SLCFc fusion protein was prepared and its chemotaxis of lymphocytes was detected by chemotaxis assay. Implanted melanoma mouse models were established and randomly divided into 4 groups: control group, SLCFc group, CpGODN group, and SLCFc+CpGODN group. The growth of implanted tumors in each group was observed after treatment. Subtype and infiltration of lymphocytes in implanted tumor tissues were examined by flow cytometry. Results:SLCFc protein was successfully prepared, and it dosedependently attracted lymphocytes (0.03, 0.3, and 3 μg/L). Intratumor injection SLCFc and CpGODN alone or in combination significantly inhibited growth of B16implanted tumors. Tumor size in SLCFc+CpGODN group was significantly smaller than that in control group (P<0.01), and animals in SLCFc+CpGODN group survived longer. Tumorinfiltrated CD4+ T, CD8+ T, and dendritic cells (DCs) in SLCFc+CpGODN group were markedly increased as compared with those in control group (P<0.05), and tumor draining lymph nodes were dramatically enlarged. Conclusion:SLC combined with CpGODN can inhibit the growth of implanted melanoma by attracting CD4+ T and CD8+ T and promoting proliferation of DCs.
Objective:To study the therapeutic effect of secondary lymphoid tissue chemokine (SLC) combined with CpG oligodeoxynucleotide (CpGODN) in treatment of implanted mouse melanoma and the possible mechanism. Methods:SLCFc fusion protein was prepared and its chemotaxis of lymphocytes was detected by chemotaxis assay. Implanted melanoma mouse models were established and randomly divided into 4 groups: control group, SLCFc group, CpGODN group, and SLCFc+CpGODN group. The growth of implanted tumors in each group was observed after treatment. Subtype and infiltration of lymphocytes in implanted tumor tissues were examined by flow cytometry. Results:SLCFc protein was successfully prepared, and it dosedependently attracted lymphocytes (0.03, 0.3, and 3 μg/L). Intratumor injection SLCFc and CpGODN alone or in combination significantly inhibited growth of B16implanted tumors. Tumor size in SLCFc+CpGODN group was significantly smaller than that in control group (P<0.01), and animals in SLCFc+CpGODN group survived longer. Tumorinfiltrated CD4+ T, CD8+ T, and dendritic cells (DCs) in SLCFc+CpGODN group were markedly increased as compared with those in control group (P<0.05), and tumor draining lymph nodes were dramatically enlarged. Conclusion:SLC combined with CpGODN can inhibit the growth of implanted melanoma by attracting CD4+ T and CD8+ T and promoting proliferation of DCs.
ZHANG Xiao-min
,
ZHANG Tang-de
,
BAO Chenchen
,
SONG Hua
,
LI Na
,
LIU Bin
,
HE Rong
,
LI Zhi-ming
,
CUI Da-xiang
,
REN Qiu-shi
2010, 17(1):30-35.
DOI: 10.3872/j.issn.1007-385X.2010.1.006
Abstract:
Objective:To prepare a nanoprobe, antihuman melanoma ganglioside single chain variable fragment (GD/ScFvMEL) antibody conjugated with CdTe quantum dot, and to observe its ability to specifically bind human malignant melanoma cells. Methods:The GD/ScFvMEL gene was cloned into pET32a (+), and the plasmid was then transformed into E. coli BL21 (DE3) for GD/ScFvMEL protein antibody expression. The expressed GD/ScFvMEL antibody was purified by denaturing method and further refolded by modified dialysis method. The purified GD/ScFvMEL antibody was analyzed by SDSPAGE. The GD/ScFvMELQDs nanoprobe was prepared by conjugating GD/ScFvMEL antibody with CdTe quantum dot, and its specificity was observed by incubating with MGC803 cells and melanoma A375 cells. Results:The recombinant pET32aGD/ScFvMEL was constructed and confirmed by PCR, restriction endonuclease analysis and DNA sequencing. The proportion of expressed GD/ScFvMEL antibody in total bacteria proteins was about 40% as detected by SDSPAGE. The purified and refoldedGD/ScFvMEL antibody was effectively conjugated with CdTe quantum dot, and the resulting GD/ScFvMELQDs nanoprobe was successfully prepared. The GD/ScFvMELQDs nanoprobe could specifically bind melanoma A375 cells, but could not bind stomach cancer MGC803 cells. Conclusion:We have successfully prepared an antihuman melanoma ganglioside singlechain antibodyCdTe quantum dot nanoprobe, which can specifically bind melanoma cells.
Objective:To prepare a nanoprobe, antihuman melanoma ganglioside single chain variable fragment (GD/ScFvMEL) antibody conjugated with CdTe quantum dot, and to observe its ability to specifically bind human malignant melanoma cells. Methods:The GD/ScFvMEL gene was cloned into pET32a (+), and the plasmid was then transformed into E. coli BL21 (DE3) for GD/ScFvMEL protein antibody expression. The expressed GD/ScFvMEL antibody was purified by denaturing method and further refolded by modified dialysis method. The purified GD/ScFvMEL antibody was analyzed by SDSPAGE. The GD/ScFvMELQDs nanoprobe was prepared by conjugating GD/ScFvMEL antibody with CdTe quantum dot, and its specificity was observed by incubating with MGC803 cells and melanoma A375 cells. Results:The recombinant pET32aGD/ScFvMEL was constructed and confirmed by PCR, restriction endonuclease analysis and DNA sequencing. The proportion of expressed GD/ScFvMEL antibody in total bacteria proteins was about 40% as detected by SDSPAGE. The purified and refoldedGD/ScFvMEL antibody was effectively conjugated with CdTe quantum dot, and the resulting GD/ScFvMELQDs nanoprobe was successfully prepared. The GD/ScFvMELQDs nanoprobe could specifically bind melanoma A375 cells, but could not bind stomach cancer MGC803 cells. Conclusion:We have successfully prepared an antihuman melanoma ganglioside singlechain antibodyCdTe quantum dot nanoprobe, which can specifically bind melanoma cells.
2010, 17(1):36-39.
DOI: 10.3872/j.issn.1007-385X.2010.1.007
Abstract:
Objective:To construct the insulinlike growth factor binding protein 7 (IGFBP7) expression plasmid (pEGFC1IGFBP7) and to investigate the effect of IGFBP7 on the apoptosis of SKMEL28 (human malignant melanoma cell line) cells. Methods: The pEGFC1IGFBP7 plasmid was constructed; pEGFC1IGFBP7 and empty plasmids were transfected into SKMEL28 cells separately. The transfection efficiency was observed under fluorescence microscope. Apoptosis of SKMEL28 cells after transfection was detected by AnnexinFITC/PI staining. Results:The pEGFC1IGFBP7 plasmid was successfully constructed and was effectively transfected into SKMEL28 cells by Effectene reagent, with the transfection rate being 61%. The results of flow cytometry showed that pEGFC1IGFBP7 significantly induced apoptosis of SKMEL28 cells, with the apoptotic rates of pEGFC1IGFBP7, empty vector, and nontransfected plasmid groups being (28.4±2.57)%, (5.8±0.44)%, and (6.4±0.71)% 24 h after transfection, respectively (F=406.138, P<005). Conclusion:pEGFC1IGFBP7 can effectively induce apoptosis of malignant melanoma SKMEL28 cells, which provides an experimental basis for IGFBP7 genebased therapy of malignant melanoma.
Objective:To construct the insulinlike growth factor binding protein 7 (IGFBP7) expression plasmid (pEGFC1IGFBP7) and to investigate the effect of IGFBP7 on the apoptosis of SKMEL28 (human malignant melanoma cell line) cells. Methods: The pEGFC1IGFBP7 plasmid was constructed; pEGFC1IGFBP7 and empty plasmids were transfected into SKMEL28 cells separately. The transfection efficiency was observed under fluorescence microscope. Apoptosis of SKMEL28 cells after transfection was detected by AnnexinFITC/PI staining. Results:The pEGFC1IGFBP7 plasmid was successfully constructed and was effectively transfected into SKMEL28 cells by Effectene reagent, with the transfection rate being 61%. The results of flow cytometry showed that pEGFC1IGFBP7 significantly induced apoptosis of SKMEL28 cells, with the apoptotic rates of pEGFC1IGFBP7, empty vector, and nontransfected plasmid groups being (28.4±2.57)%, (5.8±0.44)%, and (6.4±0.71)% 24 h after transfection, respectively (F=406.138, P<005). Conclusion:pEGFC1IGFBP7 can effectively induce apoptosis of malignant melanoma SKMEL28 cells, which provides an experimental basis for IGFBP7 genebased therapy of malignant melanoma.
2010, 17(1):40-45.
DOI: 10.3872/j.issn.1007-385X.2010.1.008
Abstract:
Objective:To probe into the relationship between integrin β1 expression in human cervical carcinoma HCE1 multicellular spheroids (HCE1/MCS) and their invasion into human umbilical vein endothelium cells (HUVEC), so as to assess the inhibitory effect of antiintegrin β1 monoclonal antibody P5D2 on the invasion of HCE1/MCS into HUVEC. Methods:A model of HUVEC monolayer invaded by HCE1/MCS was established (in brief, the HCE1 invading model). Morphology changes of the HCE1 invading model were observed under inverted microscope and analyzed by Motic Med System after P5D2 treatment. The expressions of β1 integrin on HCE1 monolayer cells, HCE1/MCS, HVUEC monolayer cells, and P5D2treated HCE1 invading models were measured by immunocytochemistry SABC assay. Results:A HCE1 invading model was successfully established. The HCE1/MCS proliferated rapidly after culture, and on the 7th day the invading area of HCE1/MCS was 40.42 folds larger than the original HCE1/MCS. Invading areas of P5D2treated HCE1/MCS were significantly smaller than that of the control group after 1, 4, 7 day (P<0.05, or P<0.01), with the invading area after 7 days reduced by (84.68±0.08) % compared with the control group. Integrin β1 expression in HCE1/MCS was significantly higher than that in HCE1 monolayer cells (P<0.01), and the expression was negative in HUVEC. Integrin β1 expression in P5D2treated HCE1/MCS was significantly lower than that the in untreated HCE1/MCS (P<0.01). Conclusion:Upregulated expression of integrin β1 in HCE1/MCS and HCE1 invading model may be associated with their enhanced adhesion and invasion abilities. Antiintegrin β1 monoclonal antibody P5D2 can partially block the invasion of HCE1/MCS into HUVEC.
Objective:To probe into the relationship between integrin β1 expression in human cervical carcinoma HCE1 multicellular spheroids (HCE1/MCS) and their invasion into human umbilical vein endothelium cells (HUVEC), so as to assess the inhibitory effect of antiintegrin β1 monoclonal antibody P5D2 on the invasion of HCE1/MCS into HUVEC. Methods:A model of HUVEC monolayer invaded by HCE1/MCS was established (in brief, the HCE1 invading model). Morphology changes of the HCE1 invading model were observed under inverted microscope and analyzed by Motic Med System after P5D2 treatment. The expressions of β1 integrin on HCE1 monolayer cells, HCE1/MCS, HVUEC monolayer cells, and P5D2treated HCE1 invading models were measured by immunocytochemistry SABC assay. Results:A HCE1 invading model was successfully established. The HCE1/MCS proliferated rapidly after culture, and on the 7th day the invading area of HCE1/MCS was 40.42 folds larger than the original HCE1/MCS. Invading areas of P5D2treated HCE1/MCS were significantly smaller than that of the control group after 1, 4, 7 day (P<0.05, or P<0.01), with the invading area after 7 days reduced by (84.68±0.08) % compared with the control group. Integrin β1 expression in HCE1/MCS was significantly higher than that in HCE1 monolayer cells (P<0.01), and the expression was negative in HUVEC. Integrin β1 expression in P5D2treated HCE1/MCS was significantly lower than that the in untreated HCE1/MCS (P<0.01). Conclusion:Upregulated expression of integrin β1 in HCE1/MCS and HCE1 invading model may be associated with their enhanced adhesion and invasion abilities. Antiintegrin β1 monoclonal antibody P5D2 can partially block the invasion of HCE1/MCS into HUVEC.
2010, 17(1):46-50.
DOI: 10.3872/j.issn.1007-385X.2010.1.009
Abstract:
Objective:To investigate the in vitro and in vivo inhibitory effects of DC (dendritic cell)CIK (cytokineinduced killer cell) cocultured cells combined with sorafenib against hepatocellular carcinoma cell line BEL7402. Methods: DC and CIK cells were generated in vitro by stimulating human peripheral blood mononuclear cells with different cytokines, and then they were cocultured. The cytotoxicity of DCCIK cocultured cells (DCCIK) combined with sorafenib against BEL7402 cells was determined by CCK8 kit. The apoptosis of BEL7402 cells was measured by Annexin VFITC Kit. BEL7402implanted tumor model was established by subcutaneous injection in nude mouse. Tumorbearing mice were divided into normal saline control group, sorafenib group, DCCIK group and DCCIK+sorafenib group. The inhibitory effects were observed in different groups. Results: The cytotoxicity rate of BEL7402 cells in DCCIK+sorafenib group was significantly higher than those in the other two groups, with cytotoxicity rate in DCCIK+sorafenib group being (75.24±1.91)%, which was 1.8 times that in DCCIK group and 2.1 times that in sorafenib group (P<0.01). The apoptosis rate of BEL7402 cells in DCCIK+sorafenib group was significantly higher than those in the sorafenib and DCCIK groups, with the apoptosis rate in DCCIK+sorafenib group being (78.32±2.54)% (P<0.05). The volume of tumor in the combination group was significantly smaller than those in the other groups (P<0.05). In vivo results showed that DCCIK+sorafenib treatment significantly inhibited the growth of BEL7402implanted tumors, and the inhibitory rate was (83.37 ±0.16)%, which was significantly higher than those of the other groups (P<0.01). Conclusion：DCCIK cocultured cells combined with sorafenib can inhibit the growth of hepatocellular carcinoma cell line BEL7402 in vitro and in vivo. Molecular targeting therapy combined with immunotherapy may be a new way for the comprehensive treatment of hepatocellular carcinoma.
Objective:To investigate the in vitro and in vivo inhibitory effects of DC (dendritic cell)CIK (cytokineinduced killer cell) cocultured cells combined with sorafenib against hepatocellular carcinoma cell line BEL7402. Methods: DC and CIK cells were generated in vitro by stimulating human peripheral blood mononuclear cells with different cytokines, and then they were cocultured. The cytotoxicity of DCCIK cocultured cells (DCCIK) combined with sorafenib against BEL7402 cells was determined by CCK8 kit. The apoptosis of BEL7402 cells was measured by Annexin VFITC Kit. BEL7402implanted tumor model was established by subcutaneous injection in nude mouse. Tumorbearing mice were divided into normal saline control group, sorafenib group, DCCIK group and DCCIK+sorafenib group. The inhibitory effects were observed in different groups. Results: The cytotoxicity rate of BEL7402 cells in DCCIK+sorafenib group was significantly higher than those in the other two groups, with cytotoxicity rate in DCCIK+sorafenib group being (75.24±1.91)%, which was 1.8 times that in DCCIK group and 2.1 times that in sorafenib group (P<0.01). The apoptosis rate of BEL7402 cells in DCCIK+sorafenib group was significantly higher than those in the sorafenib and DCCIK groups, with the apoptosis rate in DCCIK+sorafenib group being (78.32±2.54)% (P<0.05). The volume of tumor in the combination group was significantly smaller than those in the other groups (P<0.05). In vivo results showed that DCCIK+sorafenib treatment significantly inhibited the growth of BEL7402implanted tumors, and the inhibitory rate was (83.37 ±0.16)%, which was significantly higher than those of the other groups (P<0.01). Conclusion：DCCIK cocultured cells combined with sorafenib can inhibit the growth of hepatocellular carcinoma cell line BEL7402 in vitro and in vivo. Molecular targeting therapy combined with immunotherapy may be a new way for the comprehensive treatment of hepatocellular carcinoma.
MEI Mei
,
REN Yu
,
ZHOU Xuan
,
ZHAO Jin-hui
,
WANG Fan
,
GAO Wei
,
QI Yan-bin
,
YAO Zhi
,
JIANG Ling-huo
2010, 17(1):51-56.
DOI: 10.3872/j.issn.1007-385X.2010.1.010
Abstract:
Objective:To investigate the effect of RNA interference (RNAi) targeting AKT1 and PI3K P85 on the proliferation and invasion of breast carcinoma MCF7 cells. Methods:The recombinant adenovirus expression vector, which contained short hairpin RNA (shRNA) targeting open reading frames of AKT1 and PI3K P85(rAd5siAKT1siPI3K), was transfected into human breast carcinoma MCF7 cells. AKT1 and PI3K P85 mRNA and protein expressions were detected by realtime PCR and Western blotting analysis. The expressions of PCNA, cyclinD1, and P53 were also detected by Western blotting analysis. The proliferation and apoptosis of MCF7 cells were measured by MTT, flow cytometry and 2dementinal and 3dementional matrigel assay. Results:Recombinant adenovirus vector rAd5siAKT1siPI3K dramatically downregulated AKT1 and PI3K P85 mRNA and protein expressions in MCF7 cells; the downstream factors PCNA and cyclin D1 were also downregulated, while P53 was upregulated. Growth of MCF7 cells was inhibited by over 50% in rAd5siAKT1siPI3K group as measured by MTT assay, and cell cycle was arrested in G1/G0 phase compared with untransfected and rAd5siCtrl transfected groups. Cell growth on matrigel matrix showed normal cell shapes, while the cells in rAd5siAKT1siPI3K transfected group were detached from the matrix or grew in scattered clustering patterns, forming only small aggregates. Conclusion:shRNA targeting AKT1 and PI3K P85 can significantly downregulate the expression of AKT1 and PI3K P85 in breast carcinoma MCF7 cells, and inhibit the growth of MCF7 cells in vitro.
Objective:To investigate the effect of RNA interference (RNAi) targeting AKT1 and PI3K P85 on the proliferation and invasion of breast carcinoma MCF7 cells. Methods:The recombinant adenovirus expression vector, which contained short hairpin RNA (shRNA) targeting open reading frames of AKT1 and PI3K P85(rAd5siAKT1siPI3K), was transfected into human breast carcinoma MCF7 cells. AKT1 and PI3K P85 mRNA and protein expressions were detected by realtime PCR and Western blotting analysis. The expressions of PCNA, cyclinD1, and P53 were also detected by Western blotting analysis. The proliferation and apoptosis of MCF7 cells were measured by MTT, flow cytometry and 2dementinal and 3dementional matrigel assay. Results:Recombinant adenovirus vector rAd5siAKT1siPI3K dramatically downregulated AKT1 and PI3K P85 mRNA and protein expressions in MCF7 cells; the downstream factors PCNA and cyclin D1 were also downregulated, while P53 was upregulated. Growth of MCF7 cells was inhibited by over 50% in rAd5siAKT1siPI3K group as measured by MTT assay, and cell cycle was arrested in G1/G0 phase compared with untransfected and rAd5siCtrl transfected groups. Cell growth on matrigel matrix showed normal cell shapes, while the cells in rAd5siAKT1siPI3K transfected group were detached from the matrix or grew in scattered clustering patterns, forming only small aggregates. Conclusion:shRNA targeting AKT1 and PI3K P85 can significantly downregulate the expression of AKT1 and PI3K P85 in breast carcinoma MCF7 cells, and inhibit the growth of MCF7 cells in vitro.
2010, 17(1):57-61.
DOI: 10.3872/j.issn.1007-385X.2010.1.011
Abstract:
Objective: To prepare polyDLlactidepoly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase1 (TIMP1) adenovirus, and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells. Methods:The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP1 in biodegradable PELA. The diameter of the microsphere, quantity of virus encapsulated, loading rate, and releasing kinetics were measured. HepG2 cells were infected with the microspheres; the infection efficiency was examined by fluorescent microscope; and the ultrastructure was observed by TEM. The expression of TIMP1 mRNA in HepG2 cells was examined by semiquantitative RTPCR, and the proliferation of HepG2 cells was detected by MTT assay. Results:The microsphere encapsulating recombinant TIMP1 adenovirus was successfully constructed, with its diameter, entrapment efficiency, and virus loading rate being 1.965, 60.0%, and 10.5×108/mg, respectively. About 60% of the viruses were released within 120 h, and the total releasing time was longer than 240 h. Infection with rAdTIMP1 PELA microsphere efficiently induced TIMP1 expression in HepG2 cells, and significantly inhibited the proliferation of HepG2 cells, with the inhibitory rate being 47%. Conclusion:PELA microsphere encapsulating recombinant TIMP1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for the combining macromolecular chemistry and gene therapy for treatment of hepatocellular carcinoma.
Objective: To prepare polyDLlactidepoly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase1 (TIMP1) adenovirus, and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells. Methods:The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP1 in biodegradable PELA. The diameter of the microsphere, quantity of virus encapsulated, loading rate, and releasing kinetics were measured. HepG2 cells were infected with the microspheres; the infection efficiency was examined by fluorescent microscope; and the ultrastructure was observed by TEM. The expression of TIMP1 mRNA in HepG2 cells was examined by semiquantitative RTPCR, and the proliferation of HepG2 cells was detected by MTT assay. Results:The microsphere encapsulating recombinant TIMP1 adenovirus was successfully constructed, with its diameter, entrapment efficiency, and virus loading rate being 1.965, 60.0%, and 10.5×108/mg, respectively. About 60% of the viruses were released within 120 h, and the total releasing time was longer than 240 h. Infection with rAdTIMP1 PELA microsphere efficiently induced TIMP1 expression in HepG2 cells, and significantly inhibited the proliferation of HepG2 cells, with the inhibitory rate being 47%. Conclusion:PELA microsphere encapsulating recombinant TIMP1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for the combining macromolecular chemistry and gene therapy for treatment of hepatocellular carcinoma.
2010, 17(1):62-66.
DOI: 10.3872/j.issn.1007-385X.2010.1.012
Abstract:
Objective:To explore the apoptosisinducing effect of salvianolate on hepatoma SMMC7721 cells and the underlying mechanism. Methods:SMMC7721 cells were cocultured in vitro with different concentrations (0.5, 1, 2 mg/ml) of salvianolate for 24 h. The apoptotic SMMC7721 cells were examined by flow cytometry, and the changes of mitochondrial transmembrane potential were examined by mitochondrial transmembrane potential JC1 kit. The activities of caspase8, caspase9, and caspase3 were detected by spectrophotometry in the hepatoma SMMC7721 cells after cocultured with 1 mg/ml salvianolate. The changes of apoptotic SMMC7721 cells induced by salvianolate in the presence or absence of caspase9 inhibitor or caspase3 inhibitor were measured by flow cytometry. The expressions of proapoptotic protein Bax and antiapoptotic protein Bcl2 were detected by Western blotting analysis. Results:Salvianolate significantly induced apoptosis of hepatoma SMMC7721 cells (P<0.05), and the decline of mitochondrial membrane potential increased with the increase of salvianolate concentration (P<0.05). The activities of caspase9 and caspase3, but not caspase8, were increased in hepatoma cells after treatment with 1 mg/ml salvianolate for 24 h (P<0.05). The apoptosisinducing effect of salvianolate was significantly decreased in the presence of caspase9 or caspase3 inhibitors (P<005). Western blotting results showed that salvianolate increased proapoptotic protein Bax expression and decreased antiapoptotic protein Bcl2 expression. Conclusion:Salvianolate can induce the apoptosis of human hepatoma SMMC7721 cells in a dosedependent manner, which is probably mediated by mitochondrial apoptosis pathway.
Objective:To explore the apoptosisinducing effect of salvianolate on hepatoma SMMC7721 cells and the underlying mechanism. Methods:SMMC7721 cells were cocultured in vitro with different concentrations (0.5, 1, 2 mg/ml) of salvianolate for 24 h. The apoptotic SMMC7721 cells were examined by flow cytometry, and the changes of mitochondrial transmembrane potential were examined by mitochondrial transmembrane potential JC1 kit. The activities of caspase8, caspase9, and caspase3 were detected by spectrophotometry in the hepatoma SMMC7721 cells after cocultured with 1 mg/ml salvianolate. The changes of apoptotic SMMC7721 cells induced by salvianolate in the presence or absence of caspase9 inhibitor or caspase3 inhibitor were measured by flow cytometry. The expressions of proapoptotic protein Bax and antiapoptotic protein Bcl2 were detected by Western blotting analysis. Results:Salvianolate significantly induced apoptosis of hepatoma SMMC7721 cells (P<0.05), and the decline of mitochondrial membrane potential increased with the increase of salvianolate concentration (P<0.05). The activities of caspase9 and caspase3, but not caspase8, were increased in hepatoma cells after treatment with 1 mg/ml salvianolate for 24 h (P<0.05). The apoptosisinducing effect of salvianolate was significantly decreased in the presence of caspase9 or caspase3 inhibitors (P<005). Western blotting results showed that salvianolate increased proapoptotic protein Bax expression and decreased antiapoptotic protein Bcl2 expression. Conclusion:Salvianolate can induce the apoptosis of human hepatoma SMMC7721 cells in a dosedependent manner, which is probably mediated by mitochondrial apoptosis pathway.
WANG Guihua
,
LUO Xue-lai
,
SUN Li
,
DENG Yu
,
WANG Shen
,
LI Zhao-ming
,
LI Xiao-lan
,
TAO De-ding
,
HU Jun-bo
,
GONG Jian-ping
2010, 17(1):67-70.
DOI: 10.3872/j.issn.1007-385X.2010.1.013
Abstract:
Objective: To investigate the effect of retinoidinterferoninduced mortality (GRIM-19) gene on the apoptosis of colon cancer. Methods:A GRIM19 eukaryotic expression vector (pCMV-Flag-GRIM-19) was constructed and transfected into SW480 cells. Expressions of GRIM-19 and apoptosisrelated proteins were detected by Western blotting analysis. Apoptosis of SW480 cells was measured by AnnexinV/PI assay and mitochondrial membrane potential JC-1 staining. Results:The GRIM19 eukaryotic expression vector pCMVFlagGRIM-19 was successfully constructed. Expression of GRIM19 in SW480 cells was upregulated and that of apoptosisrelated protein Bclxl was downregulated after transfection with pCMV-Flag-GRIM-19. Apoptosis rate was (7.7±1.39)% in SW480 cells transfected with pCMV-Flag empty vector and (15.0 ± 2.52)% in pCMV-Flag-GRIM-19 transfected cells (P<0.05). Mitochondrial membrane potential was decreased in (7.5±2.09)% of pCMVFlag transfected cells and (17.5±3.07)% of pCMV-Flag-GRIM-19 transfected cells (P<0.05). Conclusion:In vitro GRIM-19 transfection can effectively induce apoptosis of colon cancer SW480 cells.
Objective: To investigate the effect of retinoidinterferoninduced mortality (GRIM-19) gene on the apoptosis of colon cancer. Methods:A GRIM19 eukaryotic expression vector (pCMV-Flag-GRIM-19) was constructed and transfected into SW480 cells. Expressions of GRIM-19 and apoptosisrelated proteins were detected by Western blotting analysis. Apoptosis of SW480 cells was measured by AnnexinV/PI assay and mitochondrial membrane potential JC-1 staining. Results:The GRIM19 eukaryotic expression vector pCMVFlagGRIM-19 was successfully constructed. Expression of GRIM19 in SW480 cells was upregulated and that of apoptosisrelated protein Bclxl was downregulated after transfection with pCMV-Flag-GRIM-19. Apoptosis rate was (7.7±1.39)% in SW480 cells transfected with pCMV-Flag empty vector and (15.0 ± 2.52)% in pCMV-Flag-GRIM-19 transfected cells (P<0.05). Mitochondrial membrane potential was decreased in (7.5±2.09)% of pCMVFlag transfected cells and (17.5±3.07)% of pCMV-Flag-GRIM-19 transfected cells (P<0.05). Conclusion:In vitro GRIM-19 transfection can effectively induce apoptosis of colon cancer SW480 cells.
2010, 17(1):71-76.
DOI: 10.3872/j.issn.1007-385X.2010.1.014
Abstract:
Objective:To investigate the differentially expressed genes in osteosarcoma cell lines with various metastatic potentialities, and to screen for new candidate genes related to metastasis of osteosarcomas. Methods:The total RNAs of a lowly metastatic and a highly metastatic osteosarcoma cell lines (M6 and M8) were extracted. Differentially expressed genes in the two osteosarcoma cell lines were studied by cDNA microarray. The hybridization signals were scanned with a Generation Ⅲ array scanner and analyzed by Imagequant 5.0 software. Typical differentially expressed genes were further verified by realtime quantitative PCR. Results:There were 330 differentially expressed genes between M6 and M8 cells. In the highmetastasis M8 cells, 178 genes were upregulated and 152 genes were downregulated compared to the lowmetastasis M6 cells, with 43 extremely upregulated and 49 extremely downregulated. The differentially expressed genes were mainly associated with cell proliferation, indicating these genes might be related to the inhibition of M6 cells. Other differentially expressed genes included those associated with the regulation of gene expression and signal transduction, indicating these genes might be correlated with tumor metastasis. Conclusion:cDNA microarray shows an advantage in identifying genes associated with metastasis of osteosarcoma. In M8 subset of MG63 osteosarcoma cells，43 genes are upregulated and 49 genes are downregulated, which may be related with metastasis of osteosarcoma.
Objective:To investigate the differentially expressed genes in osteosarcoma cell lines with various metastatic potentialities, and to screen for new candidate genes related to metastasis of osteosarcomas. Methods:The total RNAs of a lowly metastatic and a highly metastatic osteosarcoma cell lines (M6 and M8) were extracted. Differentially expressed genes in the two osteosarcoma cell lines were studied by cDNA microarray. The hybridization signals were scanned with a Generation Ⅲ array scanner and analyzed by Imagequant 5.0 software. Typical differentially expressed genes were further verified by realtime quantitative PCR. Results:There were 330 differentially expressed genes between M6 and M8 cells. In the highmetastasis M8 cells, 178 genes were upregulated and 152 genes were downregulated compared to the lowmetastasis M6 cells, with 43 extremely upregulated and 49 extremely downregulated. The differentially expressed genes were mainly associated with cell proliferation, indicating these genes might be related to the inhibition of M6 cells. Other differentially expressed genes included those associated with the regulation of gene expression and signal transduction, indicating these genes might be correlated with tumor metastasis. Conclusion:cDNA microarray shows an advantage in identifying genes associated with metastasis of osteosarcoma. In M8 subset of MG63 osteosarcoma cells，43 genes are upregulated and 49 genes are downregulated, which may be related with metastasis of osteosarcoma.
2010, 17(1):77-81.
DOI: 10.3872/j.issn.1007-385X.2010.1.015
Abstract:
Objective:To construct the expression plasmid of a novel gene human NBEAL1 (neurobeachin like 1), and to study its relationship with the pathological grades of glioma. Methods:Total RNA of human glioma cell line U251 was extracted. NBEAL1 expression plasmid pGEXKG/NBEAL1 was constructed and transferred into E. coli BL21. Recombinant NBEAL1 protein was induced by IPTG and further purified by GST affinity chromatographic column. The purity of recombinant NBEAL1 protein was examined by Western blotting analysis. A NBEAL1 protein specific monoclonal antibody was prepared and was used to study the relationship of NBEAL1 expression with pathological grades of glioma. Results:The NBEAL1 gene fragment was successfully cloned into pGEXKG expression plasmid and verified by DNA sequencing. The recombinant NBEAL1 protein was expressed in inclusion bodies, with a yield of more than 30% of total bacterial proteins; the purity of purified NBEAL1 protein was above 95%. Western blotting analysis confirmed that the purified protein containing GST tag and NBEAL protein. NBEAL1 protein was lowly expressed in normal brain tissues and highly expressed in low grade glioma tissues; and the expression of NBEAL1 decreased with the increase of glioma malignancy. Conclusion:The NBEAL1 protein has been successfully cloned, expressed and purified. NBEAL1 protein expression in glioma tissues is negatively associated with the pathological grades of glioma.
Objective:To construct the expression plasmid of a novel gene human NBEAL1 (neurobeachin like 1), and to study its relationship with the pathological grades of glioma. Methods:Total RNA of human glioma cell line U251 was extracted. NBEAL1 expression plasmid pGEXKG/NBEAL1 was constructed and transferred into E. coli BL21. Recombinant NBEAL1 protein was induced by IPTG and further purified by GST affinity chromatographic column. The purity of recombinant NBEAL1 protein was examined by Western blotting analysis. A NBEAL1 protein specific monoclonal antibody was prepared and was used to study the relationship of NBEAL1 expression with pathological grades of glioma. Results:The NBEAL1 gene fragment was successfully cloned into pGEXKG expression plasmid and verified by DNA sequencing. The recombinant NBEAL1 protein was expressed in inclusion bodies, with a yield of more than 30% of total bacterial proteins; the purity of purified NBEAL1 protein was above 95%. Western blotting analysis confirmed that the purified protein containing GST tag and NBEAL protein. NBEAL1 protein was lowly expressed in normal brain tissues and highly expressed in low grade glioma tissues; and the expression of NBEAL1 decreased with the increase of glioma malignancy. Conclusion:The NBEAL1 protein has been successfully cloned, expressed and purified. NBEAL1 protein expression in glioma tissues is negatively associated with the pathological grades of glioma.
2010, 17(1):82-87.
DOI: 10.3872/j.issn.1007-385X.2010.1.016
Abstract:
Objective:To study the effect of wortmannin (WM), a PI3K/Akt inhibitor, on the proliferation and apoptosis of leukemia cells and the possible mechanism. Methods:Human leukemia cell line K562 was treated with different concentrations of WM. The proliferation of K562 cells was examined by MTT assay. DNA damage in K562 cells was examined by single cell gel electrophoresis assay, and apoptosis of K562 cells was detected by Annexin VFITC/PI doublestaining. The expressions of total Akt, phosphorateAkt (pAkt), and NFκB p65 mRNA and protein were detected by RTPCR and Western blotting, respectively. Results:WM inhibited the proliferation of K562 cells in a dose and timedependent manner, with the IC50 value of 24 h being 25 nmol/L. WM also induced apoptosis of K562 cells in a dosedependent manner. DNA damage in K562 cells was demonstrated by appearance of comet tail after treatment with WM, with the rate of DNA tail and the tail length being significantly higher than those in the control group (P<0.01). WM dosedependently inhibited PAkt and NFκB p65, but not the total Akt, mRNA and protein expressions. Conclusion:WM can inhibit proliferation and induce apoptosis of K562 cells in a dose and timedependent manner, probably through downregulation of phosphorate PI3K/Akt signal pathway and NFκB expression.
Objective:To study the effect of wortmannin (WM), a PI3K/Akt inhibitor, on the proliferation and apoptosis of leukemia cells and the possible mechanism. Methods:Human leukemia cell line K562 was treated with different concentrations of WM. The proliferation of K562 cells was examined by MTT assay. DNA damage in K562 cells was examined by single cell gel electrophoresis assay, and apoptosis of K562 cells was detected by Annexin VFITC/PI doublestaining. The expressions of total Akt, phosphorateAkt (pAkt), and NFκB p65 mRNA and protein were detected by RTPCR and Western blotting, respectively. Results:WM inhibited the proliferation of K562 cells in a dose and timedependent manner, with the IC50 value of 24 h being 25 nmol/L. WM also induced apoptosis of K562 cells in a dosedependent manner. DNA damage in K562 cells was demonstrated by appearance of comet tail after treatment with WM, with the rate of DNA tail and the tail length being significantly higher than those in the control group (P<0.01). WM dosedependently inhibited PAkt and NFκB p65, but not the total Akt, mRNA and protein expressions. Conclusion:WM can inhibit proliferation and induce apoptosis of K562 cells in a dose and timedependent manner, probably through downregulation of phosphorate PI3K/Akt signal pathway and NFκB expression.
2010, 17(1):88-92.
DOI: 10.3872/j.issn.1007-385X.2010.1.017
Abstract:
Objective:To investigate the effect of nerve growth factorβ(NGF-β) on the proliferation and cell cycle of human pancreatic cancer MIA PaCa-2 cells. Methods:MIA PaCa2 cells were treated with different concentrations of NGFβ and K252a (inhibitor of NGF-β receptor TrKA) alone or in combination. Clone forming rate, proliferation, and cell cycle of MIA PaCa2 cells treated with different strategies were examined by clone formation assay, MTT, and flow cytometry, respectively. Results:NGF-β significantly increased the clone formation and proliferation of MIA PaCa-2 cells (P<0.05, P<0.01). K252a significantly inhibited the proliferation of MIA PaCa-2 cells (P<0.05), while NGF-β combined with K252a had no significant effect on the proliferation of MIA PaCa-2 cells. NGF-β arrested MIA PaCa-2 cell cycle in S phase, K252a arrested cell cycle in G0/G1 phase, and NGF-β combined with K252a arrested cell cycle in S phase. Conclusion:NGF-β can enhance the proliferation of pancreatic carcinoma MIA PaCa-2 cells.
Objective:To investigate the effect of nerve growth factorβ(NGF-β) on the proliferation and cell cycle of human pancreatic cancer MIA PaCa-2 cells. Methods:MIA PaCa2 cells were treated with different concentrations of NGFβ and K252a (inhibitor of NGF-β receptor TrKA) alone or in combination. Clone forming rate, proliferation, and cell cycle of MIA PaCa2 cells treated with different strategies were examined by clone formation assay, MTT, and flow cytometry, respectively. Results:NGF-β significantly increased the clone formation and proliferation of MIA PaCa-2 cells (P<0.05, P<0.01). K252a significantly inhibited the proliferation of MIA PaCa-2 cells (P<0.05), while NGF-β combined with K252a had no significant effect on the proliferation of MIA PaCa-2 cells. NGF-β arrested MIA PaCa-2 cell cycle in S phase, K252a arrested cell cycle in G0/G1 phase, and NGF-β combined with K252a arrested cell cycle in S phase. Conclusion:NGF-β can enhance the proliferation of pancreatic carcinoma MIA PaCa-2 cells.
2010, 17(1):93-94.
DOI: 10.3872/j.issn.1007-385X.2010.1.018
Abstract:
目的：观察caspase-3、P73、CDK4在口腔黏膜鳞状细胞癌平阳霉素治疗前后的表达及其临床意义。方法：临床标本取自河北医科大学第四医院口腔科1988年至2005年的23例病理学确诊的口腔鳞状细胞癌患者。流式细胞术检测caspase-3、P73、CDK4在口腔黏膜鳞状细胞癌中的表达。结果：Caspase-3在口腔鳞癌中的表达水平显著性低于正常组织(P<0.05)，平阳霉素化疗后表达显著性增高(P<0.05)。P73表达量在口腔鳞状细胞癌中明显升高，平阳霉素化疗后显著性下降(P<0.05)。口腔鳞状细胞癌中CDK4表达明显增高，平阳霉素化疗后显著性下降(P<0.05)。结论：Caspase-3、P73、CDK4在口腔黏膜中的异常表达与口腔鳞癌的发生有关，平阳霉素可能通过调节caspase-3、P73、CDK4的表达起到抗癌作用。
目的：观察caspase-3、P73、CDK4在口腔黏膜鳞状细胞癌平阳霉素治疗前后的表达及其临床意义。方法：临床标本取自河北医科大学第四医院口腔科1988年至2005年的23例病理学确诊的口腔鳞状细胞癌患者。流式细胞术检测caspase-3、P73、CDK4在口腔黏膜鳞状细胞癌中的表达。结果：Caspase-3在口腔鳞癌中的表达水平显著性低于正常组织(P<0.05)，平阳霉素化疗后表达显著性增高(P<0.05)。P73表达量在口腔鳞状细胞癌中明显升高，平阳霉素化疗后显著性下降(P<0.05)。口腔鳞状细胞癌中CDK4表达明显增高，平阳霉素化疗后显著性下降(P<0.05)。结论：Caspase-3、P73、CDK4在口腔黏膜中的异常表达与口腔鳞癌的发生有关，平阳霉素可能通过调节caspase-3、P73、CDK4的表达起到抗癌作用。
2010, 17(1):95-98.
DOI: 10.3872/j.issn.1007-385X.2010.1.019
Abstract:
目的：评价术前应用IL-2对进展期胃癌的有效性。方法：采用Cochrane系统评价方法，检索Cochrane图书馆临床对照试验文献库、MEDLINE、EMbase、中国生物医学文献数据库、中国期刊全文数据库、中国科技期刊全文数据库，并辅以手工检索和其他检索，收集国内、外关于胃癌切除术前接受IL2治疗组与单纯手术组治疗的随机对照试验和半随机对照试验资料，限制语种为中、英文。事先制定资料提取表，2位研究者独立提取纳入研究的有效信息并严格评价纳入资料的质量，对具有同质性的研究，采用Rev Man 4.2.10软件进行Meta分析。结果：共3个随机对照试验153例患者纳入研究，其中IL-2治疗组75例患者，单纯手术组78例患者。3个研究均采用了随机方法，且对不良反应事件按WHO标准进行分级，最高为Ⅱ级。Meta分析结果显示，IL-2治疗组与单纯手术组在CD4/CD8比值、术后并发症发生率和无瘤生存率的OR值和95% CI分别为0.10（0.05,0.22）、0.13（0.05,0.35）和4.19（2.04,8.61），差异有统计学意义。结论：进展期胃癌患者术前应用IL2有明显疗效，但仍需大样本、高质量的临床试验进一步验证。
目的：评价术前应用IL-2对进展期胃癌的有效性。方法：采用Cochrane系统评价方法，检索Cochrane图书馆临床对照试验文献库、MEDLINE、EMbase、中国生物医学文献数据库、中国期刊全文数据库、中国科技期刊全文数据库，并辅以手工检索和其他检索，收集国内、外关于胃癌切除术前接受IL2治疗组与单纯手术组治疗的随机对照试验和半随机对照试验资料，限制语种为中、英文。事先制定资料提取表，2位研究者独立提取纳入研究的有效信息并严格评价纳入资料的质量，对具有同质性的研究，采用Rev Man 4.2.10软件进行Meta分析。结果：共3个随机对照试验153例患者纳入研究，其中IL-2治疗组75例患者，单纯手术组78例患者。3个研究均采用了随机方法，且对不良反应事件按WHO标准进行分级，最高为Ⅱ级。Meta分析结果显示，IL-2治疗组与单纯手术组在CD4/CD8比值、术后并发症发生率和无瘤生存率的OR值和95% CI分别为0.10（0.05,0.22）、0.13（0.05,0.35）和4.19（2.04,8.61），差异有统计学意义。结论：进展期胃癌患者术前应用IL2有明显疗效，但仍需大样本、高质量的临床试验进一步验证。
2010, 17(1):99-103.
DOI: 10.3872/j.issn.1007-385X.2010.1.020
Abstract:
DNA, as the material basis of all living cells, triggers innate immune responses through TLR9 and other cytosolic recognition receptors. In recent years, the research progress of TLR9 is mainly manifested by the following four aspects: (1) the determinants of TLR9 interacting with its ligands; (2) the mechanisms and the importance of TLR9 translocation from the endoplasmic reticulum to the endosome; (3) the roles of the endosomal acidification and maturation, and subsequent TLR9 cleavage in TLR9 signal transduction pathway; and (4) the possible mechanisms by which the organism distinguish self DNA from microbial DNA. Meanwhile, a series of experiments on TLR9 antagonists and TLR9 deficient mice confirmed the presence of TLR9independent cytosolic DNA sensors. So far, three TLR9independent DNA sensors have been found, and they are DAI, AIM2, and RNA polymerase Ⅲ.
DNA, as the material basis of all living cells, triggers innate immune responses through TLR9 and other cytosolic recognition receptors. In recent years, the research progress of TLR9 is mainly manifested by the following four aspects: (1) the determinants of TLR9 interacting with its ligands; (2) the mechanisms and the importance of TLR9 translocation from the endoplasmic reticulum to the endosome; (3) the roles of the endosomal acidification and maturation, and subsequent TLR9 cleavage in TLR9 signal transduction pathway; and (4) the possible mechanisms by which the organism distinguish self DNA from microbial DNA. Meanwhile, a series of experiments on TLR9 antagonists and TLR9 deficient mice confirmed the presence of TLR9independent cytosolic DNA sensors. So far, three TLR9independent DNA sensors have been found, and they are DAI, AIM2, and RNA polymerase Ⅲ.
2010, 17(1):104-108.
DOI: 10.3872/j.issn.1007-385X.2010.1.021
Abstract:
2010, 17(1):109-114.
DOI: 10.3872/j.issn.1007-385X.2010.1.022
Abstract:
miRNA is a kind of endogenous noncoding short RNA. Mature miRNA was formed through the process of shearing and transporting after genetic transcription. miRNA exhibits many important biological functions through regulating expression and translation of target mRNAs. Different miRNAs may act as oncogenes or antioncogenes, and have tissue specificity. The progress of the tumorigenesis is usually accompanied by expressionprofile changes of miRNAs. MiRNA regulates many tumor biological behaviors such as differentiation, proliferation, apoptosis, invasion, metastasis, and drug resistance of tumors. Furthermore, some miRNAs have clinical significance in predicting prognosis of tumor patients.
miRNA is a kind of endogenous noncoding short RNA. Mature miRNA was formed through the process of shearing and transporting after genetic transcription. miRNA exhibits many important biological functions through regulating expression and translation of target mRNAs. Different miRNAs may act as oncogenes or antioncogenes, and have tissue specificity. The progress of the tumorigenesis is usually accompanied by expressionprofile changes of miRNAs. MiRNA regulates many tumor biological behaviors such as differentiation, proliferation, apoptosis, invasion, metastasis, and drug resistance of tumors. Furthermore, some miRNAs have clinical significance in predicting prognosis of tumor patients.
联系方式
- 《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
- 邮编：200433
- 电话：021-81871002-22
- 电子邮箱：cjcb@biother.cn
- 网址:http://www.biother.cn
- 刊号：ISSN 1007-385X
- CN 31-1725/R
- 国内定价： ￥20元/册
您是第位访问者
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.