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Volume 18,Issue 6,2011 Table of Contents
2011, 18(6):581-587.
DOI: 10.3872/j.issn.1007-385X.2011.6.001
Abstract:
Primary liver cancer is one of the most common and aggressive malignant diseases in China. The traditional therapeutic strategies to live cancer most focus on cancer cells only, but more and more researches have shown that the immune system and tumor microenvironment are also important to hepatocellular carcinogenesis. In this paper, we focus on the immune regulation to liver cancer and mainly talk about the function of tumor microenvironment, such as lymphocytes, inflammatory cells, cytokines and chemokines in hepatocellular carcinogenesis. We attempte to briefly elucidate the double-edged regulation functions of immune system on liver cancer in cellular and molecular levels. Meanwhile, we also emphasize the anti-tumor effects and potential clinical applications of these immune regulators. We believe that the further understanding of immune regulation in liver cancer will help us to discover the mechanism of carcinogenesis and explore new drug for liver cancer immunotherapy.
Primary liver cancer is one of the most common and aggressive malignant diseases in China. The traditional therapeutic strategies to live cancer most focus on cancer cells only, but more and more researches have shown that the immune system and tumor microenvironment are also important to hepatocellular carcinogenesis. In this paper, we focus on the immune regulation to liver cancer and mainly talk about the function of tumor microenvironment, such as lymphocytes, inflammatory cells, cytokines and chemokines in hepatocellular carcinogenesis. We attempte to briefly elucidate the double-edged regulation functions of immune system on liver cancer in cellular and molecular levels. Meanwhile, we also emphasize the anti-tumor effects and potential clinical applications of these immune regulators. We believe that the further understanding of immune regulation in liver cancer will help us to discover the mechanism of carcinogenesis and explore new drug for liver cancer immunotherapy.
2011, 18(6):588-596.
DOI: 10.3872/j.issn.1007-385X.2011.6.002
Abstract:
Lysophosphatidic acid (LPA) is a naturally occurring phospholipid with diverse effects on various cells, ranging from cellular morphology alterations to cellular function changes such as induction of cell proliferation, survival, drug resistance and motility. Like many other biomediators, LPA interacts with cells through specific cell surface receptors (G protein-coupled receptors). Edg-2/LPA1, Edg-4/LPA2 and Edg-7/LPA3, named as endothelial differentiation gene or lysophospholipidic receptor subfamily (Edg/LPA subfamily), are three most common LPA receptors. LPA plays a critical role as a general growth, survival and pro-angiogenic factor in the regulation of pathophysiological processes in vivo and in vitro. Recent reports in the literature suggest that abnormalities in LPA metabolism and Edg/LPA receptors function in cancer patients may contribute to the development and progression of the disease. Thus, LPA and its receptors might be potential targets for clinical cancer diagnosis and therapy. Herein we review the function and mechanism of LPA and its receptors in the development and progression of tumors with focus on human pancreatic cancer, and also clinical diagnosis and treatment has been evaluated.
Lysophosphatidic acid (LPA) is a naturally occurring phospholipid with diverse effects on various cells, ranging from cellular morphology alterations to cellular function changes such as induction of cell proliferation, survival, drug resistance and motility. Like many other biomediators, LPA interacts with cells through specific cell surface receptors (G protein-coupled receptors). Edg-2/LPA1, Edg-4/LPA2 and Edg-7/LPA3, named as endothelial differentiation gene or lysophospholipidic receptor subfamily (Edg/LPA subfamily), are three most common LPA receptors. LPA plays a critical role as a general growth, survival and pro-angiogenic factor in the regulation of pathophysiological processes in vivo and in vitro. Recent reports in the literature suggest that abnormalities in LPA metabolism and Edg/LPA receptors function in cancer patients may contribute to the development and progression of the disease. Thus, LPA and its receptors might be potential targets for clinical cancer diagnosis and therapy. Herein we review the function and mechanism of LPA and its receptors in the development and progression of tumors with focus on human pancreatic cancer, and also clinical diagnosis and treatment has been evaluated.
2011, 18(6):597-604.
DOI: 10.3872/j.issn.1007-385X.2011.6.003
Abstract:
Objective : To evaluate the regulation of hypoxia microenvironment on cytotoxicity effect of peripheral blood lymphocyte (PBL) activated by the superantigen of toxic shock syndrome toxin-1 (TSST-1) against CEA+ human colon cancer cell line LoVo. Methods: The recombinant retroviral vector pLEGFP-N1-5HRE-CEAp-TSST-1-linker-CD80TM (pLEGFP-N1-5HCTC) containing a CEA promoter and 5 copies of the hypoxia-response elements (HRE) enhancer, which was constructed in our previous study, was packaged and collected. pLEGFP-N1-5HCTC was then transducted into CEA+LoVo cells or CEA- human cervical carcinoma HeLa cells, and LoVo or HeLa cells with stable expression of transmembrane superantigen fusion gene TSST-1-linker-CD80TM protein (TC fusion protein) were obtained. Hypoxia microenvironment was simulated by CoCl2, and RT-PCR and Western blotting were employed to examine the expression of TSST-1 regulated by hypoxia. Human PBL was extracted and co-cultured with transgene tumor cells infected with pLEGFP-N1-5HCTC retrovirus; 3H-TdR assay was employed to detect the proliferation of PBL regulated by hypoxia; and the cytotocixity effect of PBL regulated by hypoxia against the transgene tumor cells was detected by MTT assay. Results: CEA+LoVo cells infected with recombinant retrovirus pLEGFP-N1-5HCTC were obtained (5HCTC/LoVo). The expression of TSST-1 mRNA and protein in 5HCTC/LoVo cells was further increased with hypoxia, which was confirmed by RT-PCR and Western blotting. There was no TSST-1 expression in CEA-HeLa cells under either hypoxic condition or normoxic condition. The proliferation of PBL activated by 5HCTC/LoVo cells was increased with hypoxia (7.3×103 vs 3.1×103 cpm, P<005); and the cytotocixity of PBL on 5HCTC/LoVo cells was more effective in hypoxia condition than in normoxic condition (82.69% vs 53.50%, P＜0.01). There was no proliferation of PBL activated by CEA-HeLa cells, and PBL also could not inhibit the proliferation of CEA-HeLa cells. Conclusion: Hypoxia microenvironment can increase the cytotocixity effect of targeting expressed superantigen TSST-1 on CEA+LoVo cells.
Objective : To evaluate the regulation of hypoxia microenvironment on cytotoxicity effect of peripheral blood lymphocyte (PBL) activated by the superantigen of toxic shock syndrome toxin-1 (TSST-1) against CEA+ human colon cancer cell line LoVo. Methods: The recombinant retroviral vector pLEGFP-N1-5HRE-CEAp-TSST-1-linker-CD80TM (pLEGFP-N1-5HCTC) containing a CEA promoter and 5 copies of the hypoxia-response elements (HRE) enhancer, which was constructed in our previous study, was packaged and collected. pLEGFP-N1-5HCTC was then transducted into CEA+LoVo cells or CEA- human cervical carcinoma HeLa cells, and LoVo or HeLa cells with stable expression of transmembrane superantigen fusion gene TSST-1-linker-CD80TM protein (TC fusion protein) were obtained. Hypoxia microenvironment was simulated by CoCl2, and RT-PCR and Western blotting were employed to examine the expression of TSST-1 regulated by hypoxia. Human PBL was extracted and co-cultured with transgene tumor cells infected with pLEGFP-N1-5HCTC retrovirus; 3H-TdR assay was employed to detect the proliferation of PBL regulated by hypoxia; and the cytotocixity effect of PBL regulated by hypoxia against the transgene tumor cells was detected by MTT assay. Results: CEA+LoVo cells infected with recombinant retrovirus pLEGFP-N1-5HCTC were obtained (5HCTC/LoVo). The expression of TSST-1 mRNA and protein in 5HCTC/LoVo cells was further increased with hypoxia, which was confirmed by RT-PCR and Western blotting. There was no TSST-1 expression in CEA-HeLa cells under either hypoxic condition or normoxic condition. The proliferation of PBL activated by 5HCTC/LoVo cells was increased with hypoxia (7.3×103 vs 3.1×103 cpm, P<005); and the cytotocixity of PBL on 5HCTC/LoVo cells was more effective in hypoxia condition than in normoxic condition (82.69% vs 53.50%, P＜0.01). There was no proliferation of PBL activated by CEA-HeLa cells, and PBL also could not inhibit the proliferation of CEA-HeLa cells. Conclusion: Hypoxia microenvironment can increase the cytotocixity effect of targeting expressed superantigen TSST-1 on CEA+LoVo cells.
2011, 18(6):605-610.
DOI: 10.3872/j.issn.1007-385X.2011.6.004
Abstract:
Objective : To investigate the effect of EphB2 on proliferation and apoptosis of human pancreatic cancer CFPAC-1 cells by target-silencing EphB2 expression using lentivirus-mediated RNA interference technique. Methods: The recombinant interfering plasmid targeting EphB2 gene was constructed and transfected into 293T cells by LipofectamineTM 2000, and the most effective interfering sequence was screened from inhibiting expression of EphB2 protein. The effective interference sequence was cloned into the pLentiGFP vector to construct lentivirus pLentiGFP-EphB2, and then transfected into CFPAC-1 cells. Stable cell line with down-regulated expression of EphB2 protein was obtained. Real-time PCR and Western blotting analysis were performed to determine the expression of EphB2 at mRNA and protein levels. Moreover, the effect of downregulation of EphB2 on proliferation and apoptosis of CFPAC-1 cells was detected by CCK8 and flow cytometry assay, respectively. Results: Recombinant lentivirus pLentiGFP-EphB2 was successfully constructed and stably transfected CFPAC-1 cells (CFPAC-1 EphB2 RNAi cells) were obtained. EphB2 expression at mRNA and protein levels in CFPAC-1 EphB2 RNAi cells were significantly lower than those at blank group, with silencing rate of EphB2 was 63% in the pLentiGFP-EphB2 group. In addition, the proliferation of CFPAC-1 EphB2 RNAi cells was increased more than those of the other two groups (1.89±0.17 vs 1.63±0.13, 1.71±0.22, P<0.05) and apoptosis was decreased \[(7.02±0.01)% vs (13.37±0.02)%, (15.71±0.02)%, P<0.05). Conclusion: pLentiGFP-EphB2 can effectively silence EphB2 expression in pancreatic cancer CFPAC-1 cells and then promote the proliferation and inhibit cell apoptosis.
Objective : To investigate the effect of EphB2 on proliferation and apoptosis of human pancreatic cancer CFPAC-1 cells by target-silencing EphB2 expression using lentivirus-mediated RNA interference technique. Methods: The recombinant interfering plasmid targeting EphB2 gene was constructed and transfected into 293T cells by LipofectamineTM 2000, and the most effective interfering sequence was screened from inhibiting expression of EphB2 protein. The effective interference sequence was cloned into the pLentiGFP vector to construct lentivirus pLentiGFP-EphB2, and then transfected into CFPAC-1 cells. Stable cell line with down-regulated expression of EphB2 protein was obtained. Real-time PCR and Western blotting analysis were performed to determine the expression of EphB2 at mRNA and protein levels. Moreover, the effect of downregulation of EphB2 on proliferation and apoptosis of CFPAC-1 cells was detected by CCK8 and flow cytometry assay, respectively. Results: Recombinant lentivirus pLentiGFP-EphB2 was successfully constructed and stably transfected CFPAC-1 cells (CFPAC-1 EphB2 RNAi cells) were obtained. EphB2 expression at mRNA and protein levels in CFPAC-1 EphB2 RNAi cells were significantly lower than those at blank group, with silencing rate of EphB2 was 63% in the pLentiGFP-EphB2 group. In addition, the proliferation of CFPAC-1 EphB2 RNAi cells was increased more than those of the other two groups (1.89±0.17 vs 1.63±0.13, 1.71±0.22, P<0.05) and apoptosis was decreased \[(7.02±0.01)% vs (13.37±0.02)%, (15.71±0.02)%, P<0.05). Conclusion: pLentiGFP-EphB2 can effectively silence EphB2 expression in pancreatic cancer CFPAC-1 cells and then promote the proliferation and inhibit cell apoptosis.
QIAN Li
,
LU Jia-hui
,
FU Yi
,
ZHENG Yue-juan
,
TONG Da-ke
,
GONG Wei-juan
,
PAN Xing-yuan
,
JI Ming-chun
2011, 18(6):611-616.
DOI: 10.3872/j.issn.1007-385X.2011.6.005
Abstract:
Objective : To study the effects of membrane-bound IL-15 in combination with retinoic acid early transcript 1ε (RAE1ε) on cytotoxicity of NK cells. Methods: Three derivatives of mouse pro-B lymphocyte cell line BaF3, expressing membrane-bound IL-15 (termed BaF3/mb15), or RAE1ε (termed BaF3/RAE), or both membrane-bound IL-15 and RAE1ε (termed BaF3/mb15/RAE) were respectively constructed in the previous study. Mouse NK cells were isolated and stimulated with irradiated BaF3 derivatives. Phenotype analysis of NK cells was performed by flow cytometry. Meanwhile, perforin and granzyme B expressions were detected in NK cells by intracellular staining; the cytotoxicity of NK cells against YAC1 lymphoma target cells was evaluated by flow cytometry. Results: NK cells stimulated with BaF3-mb15-RAE cells expressed higher levels of CD25, CD44, FasL and CD107a compared to NK cells stimulated with BaF3/mb15 or BaF3/RAE cells. However, BaF3-mb15-RAE cells showed no effects on perforin and granzyme B production of NK cells. When the effector to target ratio was 20∶1, the cytotoxicity rates of BaF3/mb15-stimulated NK cells, BaF3/RAE-stimulated NK cells and BaF3/mb15/RAE-stimulated NK cells against YAC1 cells were (39.7±2.9)%, (45.3±23)% and (59.0±6.9)%, respectively, and significantly increased compared with that of BaF3-stimulated NK cells （\[28.3±1.5\]%, P<0.01). Furthermore, BaF3/mb15/RAE-stimulated NK cells exhibited higher cytotoxicity on YAC1 cells than BaF3/mb15-stimulated or BaF3/RAE-stimulated NK cells (P<0.05). Conclusion: IL-15 combined with RAE1ε promotes the activation and cytotoxicity of NK cells.
Objective : To study the effects of membrane-bound IL-15 in combination with retinoic acid early transcript 1ε (RAE1ε) on cytotoxicity of NK cells. Methods: Three derivatives of mouse pro-B lymphocyte cell line BaF3, expressing membrane-bound IL-15 (termed BaF3/mb15), or RAE1ε (termed BaF3/RAE), or both membrane-bound IL-15 and RAE1ε (termed BaF3/mb15/RAE) were respectively constructed in the previous study. Mouse NK cells were isolated and stimulated with irradiated BaF3 derivatives. Phenotype analysis of NK cells was performed by flow cytometry. Meanwhile, perforin and granzyme B expressions were detected in NK cells by intracellular staining; the cytotoxicity of NK cells against YAC1 lymphoma target cells was evaluated by flow cytometry. Results: NK cells stimulated with BaF3-mb15-RAE cells expressed higher levels of CD25, CD44, FasL and CD107a compared to NK cells stimulated with BaF3/mb15 or BaF3/RAE cells. However, BaF3-mb15-RAE cells showed no effects on perforin and granzyme B production of NK cells. When the effector to target ratio was 20∶1, the cytotoxicity rates of BaF3/mb15-stimulated NK cells, BaF3/RAE-stimulated NK cells and BaF3/mb15/RAE-stimulated NK cells against YAC1 cells were (39.7±2.9)%, (45.3±23)% and (59.0±6.9)%, respectively, and significantly increased compared with that of BaF3-stimulated NK cells （\[28.3±1.5\]%, P<0.01). Furthermore, BaF3/mb15/RAE-stimulated NK cells exhibited higher cytotoxicity on YAC1 cells than BaF3/mb15-stimulated or BaF3/RAE-stimulated NK cells (P<0.05). Conclusion: IL-15 combined with RAE1ε promotes the activation and cytotoxicity of NK cells.
2011, 18(6):617-623.
DOI: 10.3872/j.issn.1007-385X.2011.6.006
Abstract:
Objective :To investigate the effect of microRNA-10a (miR-10a) on invasion of glioma cell line U87MG. Methods: miR-10a antisense oligodeoxynucleotides (miR-10a-anti-ODN) enveloped with liposome were transfected into glioma cell line U87MG, and nonsense miRNA and blank control were used as control groups. Flow cytometry and fluorescence microscope were employed to determine the transfection efficiency of miR-10a-anti-ODN in U87MG cells; flow cytometry was employed to detect the apoptosis and cell cycle of U87MG cells after miR-10a-anti-ODN transfection; MTT assay were used to detect the proliferation of U87MG cells; Transwell assay were applied to ascertain effect of miR-10a-anti-ODN on migration and invasion of U87MG cells; and RT-PCR and Western blotting were used to examine expressions of mRNA and protein of MMP-2, MMP-9 and MMP-14 . Results: miR-10a-anti-ODN transfection had no obvious effect on proliferation, cell cycle and apoptosis of U87MG cells; however, invasion and migration of U87MG cells was significantly decreased (invasion: \[87±7.1\] vs \[155±3.7\], \[149±6.6\], P<0.05; migration: \[78.0±5.2\] vs \[150. 3±3.7\] \[147.3±6.6\]), and mRNA and protein expressions of MMP-2, MMP-9 and MMP-14 were also significantly decreased. Conclusion: miR-10a can promote invasion of glioma cell line U87MG through upregulating MMP-2, MMP-9 and MMP-14 expressions, which might be a target in glioma therapy.
Objective :To investigate the effect of microRNA-10a (miR-10a) on invasion of glioma cell line U87MG. Methods: miR-10a antisense oligodeoxynucleotides (miR-10a-anti-ODN) enveloped with liposome were transfected into glioma cell line U87MG, and nonsense miRNA and blank control were used as control groups. Flow cytometry and fluorescence microscope were employed to determine the transfection efficiency of miR-10a-anti-ODN in U87MG cells; flow cytometry was employed to detect the apoptosis and cell cycle of U87MG cells after miR-10a-anti-ODN transfection; MTT assay were used to detect the proliferation of U87MG cells; Transwell assay were applied to ascertain effect of miR-10a-anti-ODN on migration and invasion of U87MG cells; and RT-PCR and Western blotting were used to examine expressions of mRNA and protein of MMP-2, MMP-9 and MMP-14 . Results: miR-10a-anti-ODN transfection had no obvious effect on proliferation, cell cycle and apoptosis of U87MG cells; however, invasion and migration of U87MG cells was significantly decreased (invasion: \[87±7.1\] vs \[155±3.7\], \[149±6.6\], P<0.05; migration: \[78.0±5.2\] vs \[150. 3±3.7\] \[147.3±6.6\]), and mRNA and protein expressions of MMP-2, MMP-9 and MMP-14 were also significantly decreased. Conclusion: miR-10a can promote invasion of glioma cell line U87MG through upregulating MMP-2, MMP-9 and MMP-14 expressions, which might be a target in glioma therapy.
2011, 18(6):624-629.
DOI: 10.3872/j.issn.1007-385X.2011.6.007
Abstract:
Objective :To observe the effects of Livin on biology characteristics, such as proliferation and apoptosis, of colon cancer LoVo cells by RNA interference (RNAi) targeting Livin gene. Methods: Interference vectors pSilencer4.1-L1 and pSilencer4.1-L2 targeting Livin gene were constructed and transfected into LoVo cells. Then the expression of Livin was detected by RT-PCR and Western blotting. And the apoptosis, cell cycle, colony formation, proliferation of LoVo cells, as well as sensitivity of LoVo cells to cisplatin, were detected by flow cytometry, colony formation assay, and MTT. Results: Interference vectors pSilencer4.1-L1 and pSilencer4.1-L2 were successfully constructed. pSilencer4.1-L1, but not pSilencer4.1-L2, effectively silenced the Livin mRNA and protein expression in LoVo cells (P<0.01). Compared with the control group, LoVo cells in pSilencer4.1-L1 transfection group showed an increased apoptosis rate (\[24.2±32\]% vs \[8.1±1.4\]%, P＜0.01), a decreased cell proliferation (inhibitory rate: about 70% after 72 h), a decreased colony formation rate (\[15±4.6\]% vs \[85±5.8\]%, P＜0.01), increased S phase cells (\[45.7±49\]% vs \[28.0±3.0\]%, P＜0.01), decreased G1 phase cells (\[43.0±5.2\]% vs \[62.8±5.1\]%, P＜0.01), and an increased sensitivity to cisplatin (apoptosis rate increased from \[43.4±6.9\]% to \[65.3±6.2\]%, P＜0.01). Conclusion: pSilencer4.1-L1 can effectively silence Livin gene expression in colon cancer LoVo cells, inhibit proliferation and colony formation, induce apoptosis, and enhance sensitivity of LoVo cells to cisplatin.
Objective :To observe the effects of Livin on biology characteristics, such as proliferation and apoptosis, of colon cancer LoVo cells by RNA interference (RNAi) targeting Livin gene. Methods: Interference vectors pSilencer4.1-L1 and pSilencer4.1-L2 targeting Livin gene were constructed and transfected into LoVo cells. Then the expression of Livin was detected by RT-PCR and Western blotting. And the apoptosis, cell cycle, colony formation, proliferation of LoVo cells, as well as sensitivity of LoVo cells to cisplatin, were detected by flow cytometry, colony formation assay, and MTT. Results: Interference vectors pSilencer4.1-L1 and pSilencer4.1-L2 were successfully constructed. pSilencer4.1-L1, but not pSilencer4.1-L2, effectively silenced the Livin mRNA and protein expression in LoVo cells (P<0.01). Compared with the control group, LoVo cells in pSilencer4.1-L1 transfection group showed an increased apoptosis rate (\[24.2±32\]% vs \[8.1±1.4\]%, P＜0.01), a decreased cell proliferation (inhibitory rate: about 70% after 72 h), a decreased colony formation rate (\[15±4.6\]% vs \[85±5.8\]%, P＜0.01), increased S phase cells (\[45.7±49\]% vs \[28.0±3.0\]%, P＜0.01), decreased G1 phase cells (\[43.0±5.2\]% vs \[62.8±5.1\]%, P＜0.01), and an increased sensitivity to cisplatin (apoptosis rate increased from \[43.4±6.9\]% to \[65.3±6.2\]%, P＜0.01). Conclusion: pSilencer4.1-L1 can effectively silence Livin gene expression in colon cancer LoVo cells, inhibit proliferation and colony formation, induce apoptosis, and enhance sensitivity of LoVo cells to cisplatin.
8 Adenovirus E1a gene enhances P16 gene-induced apoptosis of hepatocellular carcinoma SMMC-7721 cells
2011, 18(6):630-634.
DOI: 10.3872/j.issn.1007-385X.2011.6.008
Abstract:
Objective :To investigate the synergistic effect of anti-cancer P16 gene and adenovirus E1a gene on apoptosis and proliferation of hepatocellular carcinoma SMMC-7721 cells, and to explore the novel therapeutic strategy for tumor gene therapy. Methods: Eukaryotic expression plasmid pDC315-E1a and adenoviral vector AdCMV-P16 were constructed. The expression of P16 and E1a in SMMC-7721 cells after pDC315-E1a transfection or AdCMV-P16 infection was determined by RT-PCR and immunofluorescent labeling. SMMC-7721 cell transplanted tumors in nude mice was established. The effect of pDC315-E1a and AdCMV-P16 alone or incombination on tumor growth was observed, and the expressions of P16 and E1a in transplanted tumor tissues and apoptosis of transplanted tumor cells were determined by immunohistochemistry and TUNEL assay, respectively. Results: SMMC-7721 cells showed positive expression of both mRNA and protein levels of E1a and P16 after pDC315-E1a transfection or AdCMV-P16 infection, respectively. Compared with the control group, the apoptosis rate of transplanted tumor cells was (14.3±2.5)% (P<0.01) and tumor inhibitory rate was 361% (P<0.01) in AdCMV-P16 therapy group; those in pDC315-E1a therapy group was (8.5±2.9)% (P<0.01) and 17.1% (P>0.05); and in AdCMV-P16 combined pDC315-E1a therapy group was (27.3±6.3)% (P<0.01) and 57.2% (P<0.01), respectively. Conclusion: Adenovirus E1a gene can increase P16-induced apoptosis and cell growth inhibition in SMMC-7721 cell transplanted tumors, and thus enhance the efficacy of P16 gene therapy.
Objective :To investigate the synergistic effect of anti-cancer P16 gene and adenovirus E1a gene on apoptosis and proliferation of hepatocellular carcinoma SMMC-7721 cells, and to explore the novel therapeutic strategy for tumor gene therapy. Methods: Eukaryotic expression plasmid pDC315-E1a and adenoviral vector AdCMV-P16 were constructed. The expression of P16 and E1a in SMMC-7721 cells after pDC315-E1a transfection or AdCMV-P16 infection was determined by RT-PCR and immunofluorescent labeling. SMMC-7721 cell transplanted tumors in nude mice was established. The effect of pDC315-E1a and AdCMV-P16 alone or incombination on tumor growth was observed, and the expressions of P16 and E1a in transplanted tumor tissues and apoptosis of transplanted tumor cells were determined by immunohistochemistry and TUNEL assay, respectively. Results: SMMC-7721 cells showed positive expression of both mRNA and protein levels of E1a and P16 after pDC315-E1a transfection or AdCMV-P16 infection, respectively. Compared with the control group, the apoptosis rate of transplanted tumor cells was (14.3±2.5)% (P<0.01) and tumor inhibitory rate was 361% (P<0.01) in AdCMV-P16 therapy group; those in pDC315-E1a therapy group was (8.5±2.9)% (P<0.01) and 17.1% (P>0.05); and in AdCMV-P16 combined pDC315-E1a therapy group was (27.3±6.3)% (P<0.01) and 57.2% (P<0.01), respectively. Conclusion: Adenovirus E1a gene can increase P16-induced apoptosis and cell growth inhibition in SMMC-7721 cell transplanted tumors, and thus enhance the efficacy of P16 gene therapy.
SUN Li-chen
,
ZHANG Bai-he
,
ZHANG Qi
,
SU Chang-qing
,
LI Gen-cong
,
WU Hong-ping
,
WU Meng-chao
,
QIAN Qi-jun
2011, 18(6):635-640.
DOI: 10.3872/j.issn.1007-385X.2011.6.009
Abstract:
Objective : To investigate the cytotoxicity of the novel replicative adenovirus CNHK500-hγ, a recombinant adenovirus with the adenovirus E1A and E1B genes driven by human telomerase reverse transcriptase (hTERT) and hypoxia response element (HRE) promoters respectively and carrying the hIFN-γ, against hepatocellular carcinoma (HCC) cells in vitro. Methods: The amplification and cytotoxicity of replicating adenovirus CNHK500-hγ on two telomerase positive HCC cell lines (HepG2 and Hep3B) and one telomerase negative normal cell line (BJ) were analyzed by TCID50 and MTT assays. BJ, Hep3B and HepG2 cells were infected with CNHK500-GFP carrying green fluorescent protein and the amplification of CNHK500-GFP was observed. The expressions of hIFN-γ in cells and cell supernatants after CNHK500-hγ infection were detected by Western blotting and ELISA assays. Results: Forty-eight hours after infection, the amplification of CNHK500-GFP in HepG2 and Hep3B cells were 16 003 and 2 116 times of that in BJ cells, and the cytotoxicity ED50 of CNHK500-GFP against BJ cells was respectively 500 and 10 000 times of that against HepG2 and Hep3B cells, and superior to the positive control of replicative adenovirus ONYX-015. Furthermore, hIFN-γ expression in HepG2 and Hep3B cells after CNHK500-hγ infection was significantly higher than that after non-replicative adenovirus Ad-hγ infection (P<0.01). Conclusion: Replicative adenovirus CNHK500-hγ can specifically amplify in HCC cells and effectively express hIFN-γ gene, which holds potential for the treatment of HCC.
Objective : To investigate the cytotoxicity of the novel replicative adenovirus CNHK500-hγ, a recombinant adenovirus with the adenovirus E1A and E1B genes driven by human telomerase reverse transcriptase (hTERT) and hypoxia response element (HRE) promoters respectively and carrying the hIFN-γ, against hepatocellular carcinoma (HCC) cells in vitro. Methods: The amplification and cytotoxicity of replicating adenovirus CNHK500-hγ on two telomerase positive HCC cell lines (HepG2 and Hep3B) and one telomerase negative normal cell line (BJ) were analyzed by TCID50 and MTT assays. BJ, Hep3B and HepG2 cells were infected with CNHK500-GFP carrying green fluorescent protein and the amplification of CNHK500-GFP was observed. The expressions of hIFN-γ in cells and cell supernatants after CNHK500-hγ infection were detected by Western blotting and ELISA assays. Results: Forty-eight hours after infection, the amplification of CNHK500-GFP in HepG2 and Hep3B cells were 16 003 and 2 116 times of that in BJ cells, and the cytotoxicity ED50 of CNHK500-GFP against BJ cells was respectively 500 and 10 000 times of that against HepG2 and Hep3B cells, and superior to the positive control of replicative adenovirus ONYX-015. Furthermore, hIFN-γ expression in HepG2 and Hep3B cells after CNHK500-hγ infection was significantly higher than that after non-replicative adenovirus Ad-hγ infection (P<0.01). Conclusion: Replicative adenovirus CNHK500-hγ can specifically amplify in HCC cells and effectively express hIFN-γ gene, which holds potential for the treatment of HCC.
2011, 18(6):641-464.
DOI: 10.3872/j.issn.1007-385X.2011.6.010
Abstract:
Objective : To construct a new radiation-induced, EGR-1 promotor-regulated, human TRAIL gene containing oncolytic adenovirus Ad-EGR-TRAIL, and to investigate the cytotoxicity effect of Ad-EGR-TRAIL combined with chemotherapy on cervical cancer HeLa S3 cells. Methods: Recombinant adenovirus Ad-EGR-TRAIL was constructed. HeLa S3 cells were infected with Ad-GFP, and infection efficiency was observed. The cytotoxicity effect of Ad-EGRTRAIL, radiotherapy (RAD), and Ad-EGR-TRAIL+RAD on HeLa S3 cells, as well as on normal human cervical cells, was examined by CCK-8 method. Results: Recombinant adenovirus Ad-EGR-TRAIL was successfully constructed. Ad-EGR-TRAIL showed the highest infection efficiency at MOI=100 in HeLa S3 cells. The inhibitory rates of HeLa S3 cells were (8.07±3.02)% and (23.02±4.03)% when Ad-EGR-TRAIL or RAD was used alone; however, the inhibitory rate reached (79.77±9.15)% when Ad-EGR-TRAIL and RAD were used in combination; and normal cervical cells did not significantly respond to the combination Ad-EGR-TRAIL and RAD therapy. Conclusion: Ad-EGR-TRAIL combined with chemotherapy can significantly kill cervical cancer HeLa S3 cells.
Objective : To construct a new radiation-induced, EGR-1 promotor-regulated, human TRAIL gene containing oncolytic adenovirus Ad-EGR-TRAIL, and to investigate the cytotoxicity effect of Ad-EGR-TRAIL combined with chemotherapy on cervical cancer HeLa S3 cells. Methods: Recombinant adenovirus Ad-EGR-TRAIL was constructed. HeLa S3 cells were infected with Ad-GFP, and infection efficiency was observed. The cytotoxicity effect of Ad-EGRTRAIL, radiotherapy (RAD), and Ad-EGR-TRAIL+RAD on HeLa S3 cells, as well as on normal human cervical cells, was examined by CCK-8 method. Results: Recombinant adenovirus Ad-EGR-TRAIL was successfully constructed. Ad-EGR-TRAIL showed the highest infection efficiency at MOI=100 in HeLa S3 cells. The inhibitory rates of HeLa S3 cells were (8.07±3.02)% and (23.02±4.03)% when Ad-EGR-TRAIL or RAD was used alone; however, the inhibitory rate reached (79.77±9.15)% when Ad-EGR-TRAIL and RAD were used in combination; and normal cervical cells did not significantly respond to the combination Ad-EGR-TRAIL and RAD therapy. Conclusion: Ad-EGR-TRAIL combined with chemotherapy can significantly kill cervical cancer HeLa S3 cells.
2011, 18(6):647-652.
DOI: 10.3872/j.issn.1007-385X.2011.6.011
Abstract:
Objective :To investigate the tropism of human bone-derived mesenchymal stem cells (hBMSCs) to gastric cancer cells in vitro and in vivo and to provide evidence for MSC as the vehicle for gene therapy of gastric cancer. Methods: hBMSCs were isolated and cultured using bone marrow plating method, and identified by flow cytometry. Migration abilities of hBMSCs to gastric cancer MKN45 cells and human fibroblast (hFB) cells were examined by Transwell assay. the tumor nude mice model transplanted by MKN45 cells was established. And Lenti-EGFP infected-hBMSC or-hFB (MOI=50) cells were subcutaneously injected into tumor-bearing mice, then GFP expression in transplanted tumors and different organs were observed under a fluorescence microscope. Results: The positive expression rates of CD44 and CD105 in the third passage hBMSC were (96.7±1.84)% and (98.1±0.95)% respectively, while CD34 and CD45 were negatively expressed. Chemotaxis of hBMSC to MKN45 cells was significantly higher than that to gastric epithelial GES-1 cells (239.5±54.3 vs 43.57±4.6, 37.3±4.7; P<0.01), and chemotaxis of hBMSC to MKN45 cells was significantly higher than that of hFB cells (P>0.05). hBMSC migrating to MKN45 were significantly higher than hFB (239.5±54.3 vs 27.7±16.7, P<0.01). Compared with hFB cells, hBMSC showed obvious chemotaxis to gastric transplanted tumor tissues; in hBMSC group, GFP was highly expressed in transplanted tumor tissues, as well as in liver (20%) and lung tissues (20%), but the later were significantly lower than that in transplanted tumor tissues (P<005). Conclusion: The chemotaxis of hBMSC to gastric cancer is obvious and specific, and hBMSC may be a good vehicle for gene therapy of gastric cancer.
Objective :To investigate the tropism of human bone-derived mesenchymal stem cells (hBMSCs) to gastric cancer cells in vitro and in vivo and to provide evidence for MSC as the vehicle for gene therapy of gastric cancer. Methods: hBMSCs were isolated and cultured using bone marrow plating method, and identified by flow cytometry. Migration abilities of hBMSCs to gastric cancer MKN45 cells and human fibroblast (hFB) cells were examined by Transwell assay. the tumor nude mice model transplanted by MKN45 cells was established. And Lenti-EGFP infected-hBMSC or-hFB (MOI=50) cells were subcutaneously injected into tumor-bearing mice, then GFP expression in transplanted tumors and different organs were observed under a fluorescence microscope. Results: The positive expression rates of CD44 and CD105 in the third passage hBMSC were (96.7±1.84)% and (98.1±0.95)% respectively, while CD34 and CD45 were negatively expressed. Chemotaxis of hBMSC to MKN45 cells was significantly higher than that to gastric epithelial GES-1 cells (239.5±54.3 vs 43.57±4.6, 37.3±4.7; P<0.01), and chemotaxis of hBMSC to MKN45 cells was significantly higher than that of hFB cells (P>0.05). hBMSC migrating to MKN45 were significantly higher than hFB (239.5±54.3 vs 27.7±16.7, P<0.01). Compared with hFB cells, hBMSC showed obvious chemotaxis to gastric transplanted tumor tissues; in hBMSC group, GFP was highly expressed in transplanted tumor tissues, as well as in liver (20%) and lung tissues (20%), but the later were significantly lower than that in transplanted tumor tissues (P<005). Conclusion: The chemotaxis of hBMSC to gastric cancer is obvious and specific, and hBMSC may be a good vehicle for gene therapy of gastric cancer.
2011, 18(6):653-657.
DOI: 10.3872/j.issn.1007-385X.2011.6.012
Abstract:
Objective : To investigate the effect of canstatin on growth, metastasis and angiogenesis of transplanted Lewis lung cancer in mice. Methods: The recombinant pCMV-Script/canstatin vector or the empty vector was transfected into A549 cells by electroporation, and the positive clones were screened with G418. The expressions of canstatin mRNA and protein in transfected A549 cells were examined by RT-PCR and Western blotting, respectively. Furthermore, transplanted Lewis lung cancer mouse model was established, and therapeutic effect of supernatant of pCMV-Script/canstatin transfected A549 cells on transplanted Lewis lung cancer was observed. Microvessel density of transplanted tumors in different therapy groups was observed by immunohistochemistry. Results: Positive clones of A549 cells transfected with pCMV-Script/canstatin were successfully obtained by G418 screening, and could effectively express canstatin mRNA and protein. The tumor size of the pCMV-Script/canstatin group (1.47±0.21 cm3) was significantly smaller than that in the pCMV-Scrip group (2.43±0.15 cm3) and NS group (2.53±0.18 cm3) (P<0.01). The number of pulmonary metastatic nodes was 3.00±1.00, 7.80±1.48 and 7.60±2.41 respectively for pCMV-Script/canstatin, pCMV-Script and NS groups, so pCMV-Script/canstatin significantly inhibited metastasis of tumors (P<0.01). The amount of microvessel count (MVC) in pCMV-Script/canstatin group was markedly decreased compared to that of pCMV-Script and NS groups (P<0.01). Conclusion: The pCMV-Script/canstatin mediates expression of canstatin in A549 cells. Canstatin in A549 supernantant has a strong inhibitory effect on growth, metastasis and angiogenesis of Lewis lung carcinoma.
Objective : To investigate the effect of canstatin on growth, metastasis and angiogenesis of transplanted Lewis lung cancer in mice. Methods: The recombinant pCMV-Script/canstatin vector or the empty vector was transfected into A549 cells by electroporation, and the positive clones were screened with G418. The expressions of canstatin mRNA and protein in transfected A549 cells were examined by RT-PCR and Western blotting, respectively. Furthermore, transplanted Lewis lung cancer mouse model was established, and therapeutic effect of supernatant of pCMV-Script/canstatin transfected A549 cells on transplanted Lewis lung cancer was observed. Microvessel density of transplanted tumors in different therapy groups was observed by immunohistochemistry. Results: Positive clones of A549 cells transfected with pCMV-Script/canstatin were successfully obtained by G418 screening, and could effectively express canstatin mRNA and protein. The tumor size of the pCMV-Script/canstatin group (1.47±0.21 cm3) was significantly smaller than that in the pCMV-Scrip group (2.43±0.15 cm3) and NS group (2.53±0.18 cm3) (P<0.01). The number of pulmonary metastatic nodes was 3.00±1.00, 7.80±1.48 and 7.60±2.41 respectively for pCMV-Script/canstatin, pCMV-Script and NS groups, so pCMV-Script/canstatin significantly inhibited metastasis of tumors (P<0.01). The amount of microvessel count (MVC) in pCMV-Script/canstatin group was markedly decreased compared to that of pCMV-Script and NS groups (P<0.01). Conclusion: The pCMV-Script/canstatin mediates expression of canstatin in A549 cells. Canstatin in A549 supernantant has a strong inhibitory effect on growth, metastasis and angiogenesis of Lewis lung carcinoma.
2011, 18(6):658-662.
DOI: 10.3872/j.issn.1007-385X.2011.6.013
Abstract:
Objective : To prepare anti-cancer targeting drug paclitaxel albumin magnetic nanoparticles (MAG-TAX-NP) and iodinate oil MAG-TAX-NP, and study the efficacy of iodinate oil MAG-TAX-NP on rat liver cancer. Methods: MAG-TAX-NP was prepared by emulsion-ultrasonic-solidification heat method using albumin and nano-Fe3O4 particles as carriers, and then iodinate oil MAG-TAX-NP was constructed. The content of paclitaxel was detected by UV spectrophotometry, and the loading rate and the entrapment efficiency of MAG-TAX-NP were calculated. The morphology of MAG-TAX-NP was observed under electron microscope. Rat liver tumor model was established. In addition to the blank group, 0.2 ml of iodinate oil, iodinate oil paclitaxel, iodinate oil magnetic nanoparticle, or iodinate oil MAG-TAX-NP was separately injected into the hepatic artery of tumor-bearing rats. Tumor-bearing rats were sacrificed 14 d after injection, the liver tumors were weighed and the tumor inhibitory rates in each group were calculated. The inhibitory rate of Iodinate oil MAG-TAX-NP on liver tumors was observed under an electron microscope. Results: The prepared MAG-TAX-NP was smooth; its diameter was about 70 nm; its average loading rate was 5.62%; and its average entrapment efficiency was 8072%. The tumor inhibitory rate of iodinate oil MAG-TAX-NP treatment group was significantly increased (P<0.01), with tumor inhibitory rates of iodinate oil, iodinate oil paclitaxel, iodinate oil magnetic nanoparticle, and iodinate oil MAG-TAX-NP treatment groups being (43.2±2.24)%, (51±3.33)%, (57.4±3.66)%, and (87.4±4.11)%, respectively. Furthermore, liver tumor cells showed lots of apoptosis and necrosis after iodinate oil MAG-TAX-NP treatment. Conclusion: Iodinate oil MAG-TAX-NP can inhibit growth of rat liver tumors, which might become a novel paclitaxel formulation in treatment of liver cancers.
Objective : To prepare anti-cancer targeting drug paclitaxel albumin magnetic nanoparticles (MAG-TAX-NP) and iodinate oil MAG-TAX-NP, and study the efficacy of iodinate oil MAG-TAX-NP on rat liver cancer. Methods: MAG-TAX-NP was prepared by emulsion-ultrasonic-solidification heat method using albumin and nano-Fe3O4 particles as carriers, and then iodinate oil MAG-TAX-NP was constructed. The content of paclitaxel was detected by UV spectrophotometry, and the loading rate and the entrapment efficiency of MAG-TAX-NP were calculated. The morphology of MAG-TAX-NP was observed under electron microscope. Rat liver tumor model was established. In addition to the blank group, 0.2 ml of iodinate oil, iodinate oil paclitaxel, iodinate oil magnetic nanoparticle, or iodinate oil MAG-TAX-NP was separately injected into the hepatic artery of tumor-bearing rats. Tumor-bearing rats were sacrificed 14 d after injection, the liver tumors were weighed and the tumor inhibitory rates in each group were calculated. The inhibitory rate of Iodinate oil MAG-TAX-NP on liver tumors was observed under an electron microscope. Results: The prepared MAG-TAX-NP was smooth; its diameter was about 70 nm; its average loading rate was 5.62%; and its average entrapment efficiency was 8072%. The tumor inhibitory rate of iodinate oil MAG-TAX-NP treatment group was significantly increased (P<0.01), with tumor inhibitory rates of iodinate oil, iodinate oil paclitaxel, iodinate oil magnetic nanoparticle, and iodinate oil MAG-TAX-NP treatment groups being (43.2±2.24)%, (51±3.33)%, (57.4±3.66)%, and (87.4±4.11)%, respectively. Furthermore, liver tumor cells showed lots of apoptosis and necrosis after iodinate oil MAG-TAX-NP treatment. Conclusion: Iodinate oil MAG-TAX-NP can inhibit growth of rat liver tumors, which might become a novel paclitaxel formulation in treatment of liver cancers.
2011, 18(6):663-667.
DOI: 10.3872/j.issn.1007-385X.2011.6.014
Abstract:
Objective : To investigate the prognostic factors of large cell lung cancer (LCLC) and analyze the efficacy of different postoperative therapeutic strategies on surgery-LCLC. Methods: To collect and retrospectively analyze the clinical data of 80 surgery-LCLC cases between 2000.1 and 2009.12. The prognostic factors and efficacy of different postoperative therapeutic strategies were evaluated by univariate and multivariate analyses. Results: All the 80 cases were diagnosed as stage Ⅰ (21), stage Ⅱ (22), stage Ⅲ (28) and stage Ⅳ (9). 29 cases received no systemic treatment after surgery, 35 received routine chemotherapy, 5 received IFN-α combined with chemotherapy and 11 received cytokine-induced killer cells（CIKs）combined with chemotherapy. The overall 1-, 3- and 5-year survival rates of the 80 LCLC cases were 72.5%, 45.6%, 31.0%, respectively. Cox univariate analysis revealed that N stage (P=0.002), M stage (P<0.001), the clinical stage (P<0.001), surgical methods (P=0.001) and different postoperative therapeutic strategies (P<0.001) were prognostic factors. Cox multivariate analysis indicated that the clinical stage (P<0.001), surgical methods (P=0.034), different postoperative therapeutic therapies (P=0.001) were independent prognostic factors. Further analysis revealed that the overall survival of patients with chemotherapy alone or CIK combined with chemotherapy were significantly higher than those without any postoperative therapy (all P<0.05). After analyzing 43 phase Ⅰ/Ⅱ LCLC cases, we found that the patients undergoing CIK combined with chemotherapy had a better survival than did those without any postoperative treatment or with chemotherapy alone (P=0.004, 0.044, respectively). Analysis of phase Ⅲ/Ⅳ 37 cases revealed that the overall survivals of patients recieving chemotherapy alone or IFN-α combined with chemotherapy, CIK combined with chemotherapy were significantly higher than those without any postoperative therapy (P=0.012, 0.041, 0.011, respectively). Conclusion: The clinical stage, surgical methods, postoperative therapy strategies are independent prognostic factors for LCLC patients. For early or advanced stage LCLC cases, postoperative therapy is also required, with CIK combined chemotherapy superior to chemotherapy alone.
Objective : To investigate the prognostic factors of large cell lung cancer (LCLC) and analyze the efficacy of different postoperative therapeutic strategies on surgery-LCLC. Methods: To collect and retrospectively analyze the clinical data of 80 surgery-LCLC cases between 2000.1 and 2009.12. The prognostic factors and efficacy of different postoperative therapeutic strategies were evaluated by univariate and multivariate analyses. Results: All the 80 cases were diagnosed as stage Ⅰ (21), stage Ⅱ (22), stage Ⅲ (28) and stage Ⅳ (9). 29 cases received no systemic treatment after surgery, 35 received routine chemotherapy, 5 received IFN-α combined with chemotherapy and 11 received cytokine-induced killer cells（CIKs）combined with chemotherapy. The overall 1-, 3- and 5-year survival rates of the 80 LCLC cases were 72.5%, 45.6%, 31.0%, respectively. Cox univariate analysis revealed that N stage (P=0.002), M stage (P<0.001), the clinical stage (P<0.001), surgical methods (P=0.001) and different postoperative therapeutic strategies (P<0.001) were prognostic factors. Cox multivariate analysis indicated that the clinical stage (P<0.001), surgical methods (P=0.034), different postoperative therapeutic therapies (P=0.001) were independent prognostic factors. Further analysis revealed that the overall survival of patients with chemotherapy alone or CIK combined with chemotherapy were significantly higher than those without any postoperative therapy (all P<0.05). After analyzing 43 phase Ⅰ/Ⅱ LCLC cases, we found that the patients undergoing CIK combined with chemotherapy had a better survival than did those without any postoperative treatment or with chemotherapy alone (P=0.004, 0.044, respectively). Analysis of phase Ⅲ/Ⅳ 37 cases revealed that the overall survivals of patients recieving chemotherapy alone or IFN-α combined with chemotherapy, CIK combined with chemotherapy were significantly higher than those without any postoperative therapy (P=0.012, 0.041, 0.011, respectively). Conclusion: The clinical stage, surgical methods, postoperative therapy strategies are independent prognostic factors for LCLC patients. For early or advanced stage LCLC cases, postoperative therapy is also required, with CIK combined chemotherapy superior to chemotherapy alone.
2011, 18(6):668-671.
DOI: 10.3872/j.issn.1007-385X.2011.6.015
Abstract:
Objective : To explore the loss of heterozygosity (LOH) of microsatellite sites at chromosome 3p in ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) lesions. Methods: Forty-three paraffin blocks were obtained from patients with invasive ductal carcinoma who received surgery (Sep. 2005 to Feb. 2009, Changhai Hospital). The IDC, DCIS and normal tissues were microdissected from paraffin sections. The LOH of 4 microsatellite sites was examined with the application of capillary electrophoresis sequencing techniques. Results: In 40 breast cancer cases, the frequencies of LOH at D3S1038, D3S1295, D3S1581 and D3S3118 were 25% (8/40), 37.5% (15/40), 17.5% (7/40) and 5% (2/40), respectively. The frequency of LOH in IDC group was higher than that in DCIS group with no significant difference (47.5% vs 37.5%). Conclusion: LOH at chromosome 3P site is an early event in the development of breast ductal carcinoma. The frequency of LOH in DCIS is close to that in IDC. In the development of breast carcinoma, new MS sites have LOH.
Objective : To explore the loss of heterozygosity (LOH) of microsatellite sites at chromosome 3p in ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) lesions. Methods: Forty-three paraffin blocks were obtained from patients with invasive ductal carcinoma who received surgery (Sep. 2005 to Feb. 2009, Changhai Hospital). The IDC, DCIS and normal tissues were microdissected from paraffin sections. The LOH of 4 microsatellite sites was examined with the application of capillary electrophoresis sequencing techniques. Results: In 40 breast cancer cases, the frequencies of LOH at D3S1038, D3S1295, D3S1581 and D3S3118 were 25% (8/40), 37.5% (15/40), 17.5% (7/40) and 5% (2/40), respectively. The frequency of LOH in IDC group was higher than that in DCIS group with no significant difference (47.5% vs 37.5%). Conclusion: LOH at chromosome 3P site is an early event in the development of breast ductal carcinoma. The frequency of LOH in DCIS is close to that in IDC. In the development of breast carcinoma, new MS sites have LOH.
16 Translational journey of the first FDA-approved autologous cellular immunotherapy drug sipuleucel-T
2011, 18(6):672-677.
DOI: 10.3872/j.issn.1007-385X.2011.6.016
Abstract:
自本年度诺贝尔医学或生理学奖获得者Ralph M. Steinman于1973年发现DC及其在获得性免疫应答中关键作用以来，有关DC肿瘤疫苗的研究持续进行了数十年，直到2010年4月，美国FDA才批准了首个以DC为主要效应细胞的自体细胞免疫治疗药物sipuleucel-T（又称APC8015或Provenge)用于无症状或轻微症状的转移性去势拮抗性前列腺癌的治疗，成为自1971年理查德·尼克松颁布《国家癌症法》以来癌症研究40年中的重要事件之一。为了详细了解美国Dendreon公司为临床开发sipuleucel-T所经历的坎坷历程，本文对sipuleucel-T的制备过程和作用机制、sipuleucel-TⅠ/Ⅱ期和Ⅲ期临床试验的设计情况及研究结果、sipuleucel-T上市的竞争情况作了回顾和介绍，同时分析了临床应用sipuleucel-T所亟待解决的一些问题，如疗效评价体系的建立等，希望对致力于DC肿瘤疫苗研发乃至肿瘤免疫治疗的同行有所借鉴和启示。
自本年度诺贝尔医学或生理学奖获得者Ralph M. Steinman于1973年发现DC及其在获得性免疫应答中关键作用以来，有关DC肿瘤疫苗的研究持续进行了数十年，直到2010年4月，美国FDA才批准了首个以DC为主要效应细胞的自体细胞免疫治疗药物sipuleucel-T（又称APC8015或Provenge)用于无症状或轻微症状的转移性去势拮抗性前列腺癌的治疗，成为自1971年理查德·尼克松颁布《国家癌症法》以来癌症研究40年中的重要事件之一。为了详细了解美国Dendreon公司为临床开发sipuleucel-T所经历的坎坷历程，本文对sipuleucel-T的制备过程和作用机制、sipuleucel-TⅠ/Ⅱ期和Ⅲ期临床试验的设计情况及研究结果、sipuleucel-T上市的竞争情况作了回顾和介绍，同时分析了临床应用sipuleucel-T所亟待解决的一些问题，如疗效评价体系的建立等，希望对致力于DC肿瘤疫苗研发乃至肿瘤免疫治疗的同行有所借鉴和启示。
2011, 18(6):678-681.
DOI: 10.3872/j.issn.1007-385X.2011.6.017
Abstract:
充足的营养和能量供应是癌细胞得以无限增殖、浸润和转移的基础和前提。癌细胞的葡萄糖、氨基酸和脂肪代谢都与正常细胞不同，存在着普遍的能量代谢异常。葡萄糖是癌细胞能量供给的主要来源，癌细胞的葡萄糖转运载体和一些糖酵解的关键酶活性往往增高，线粒体的氧化磷酸化功能受损。有氧糖酵解是癌细胞能量代谢的最主要特征，即使在有氧环境下，癌细胞也优先进行糖酵解以获得能量，同时生成大量乳酸；与氧化磷酸化相比较，这是一种低效的能量代谢方式。通过优先进行糖酵解，可为癌细胞细胞器的合成提供原料，具有一定的生理意义。一些基因的异常表达和肿瘤内环境低氧状态是癌细胞优先进行糖酵解的主要原因，糖酵解的关键酶或载体有望成为对恶性肿瘤进行分子靶向治疗的重要靶点。
充足的营养和能量供应是癌细胞得以无限增殖、浸润和转移的基础和前提。癌细胞的葡萄糖、氨基酸和脂肪代谢都与正常细胞不同，存在着普遍的能量代谢异常。葡萄糖是癌细胞能量供给的主要来源，癌细胞的葡萄糖转运载体和一些糖酵解的关键酶活性往往增高，线粒体的氧化磷酸化功能受损。有氧糖酵解是癌细胞能量代谢的最主要特征，即使在有氧环境下，癌细胞也优先进行糖酵解以获得能量，同时生成大量乳酸；与氧化磷酸化相比较，这是一种低效的能量代谢方式。通过优先进行糖酵解，可为癌细胞细胞器的合成提供原料，具有一定的生理意义。一些基因的异常表达和肿瘤内环境低氧状态是癌细胞优先进行糖酵解的主要原因，糖酵解的关键酶或载体有望成为对恶性肿瘤进行分子靶向治疗的重要靶点。
2011, 18(6):682-685.
DOI: 10.3872/j.issn.1007-385X.2011.6.018
Abstract:
乙醛脱氢酶1（aldehyde dehydrogenase 1, ALDH1）是一种可以将醛类氧化为乙酸的胞质溶酶，其在胚胎干细胞和成体干细胞中表达增高，对维持干细胞（stem cell, SC)的“干性”中起着重要作用。近年发现，ALDH1在肿瘤干细胞（cancer stem cell,CSC）中表达也升高。基于ALDH1活性的Aldefluor分析法已成功分选和鉴定造血干细胞、乳腺干细胞和神经干细胞等多种SC，也已成功分选和鉴定白血病干细胞、乳腺癌干细胞、肺癌干细胞、膀胱癌干细胞和头颈部肿瘤干细胞等多种CSC。免疫组化原位染色能有效检测ALDH1活性，从而也可鉴定SC和CSC，目前正在研究将此方法应用于临床评估肿瘤患者的预后。ALDH1已成为分选和鉴定SC和CSC的通用标志物之一，是SC研究的得力工具。
乙醛脱氢酶1（aldehyde dehydrogenase 1, ALDH1）是一种可以将醛类氧化为乙酸的胞质溶酶，其在胚胎干细胞和成体干细胞中表达增高，对维持干细胞（stem cell, SC)的“干性”中起着重要作用。近年发现，ALDH1在肿瘤干细胞（cancer stem cell,CSC）中表达也升高。基于ALDH1活性的Aldefluor分析法已成功分选和鉴定造血干细胞、乳腺干细胞和神经干细胞等多种SC，也已成功分选和鉴定白血病干细胞、乳腺癌干细胞、肺癌干细胞、膀胱癌干细胞和头颈部肿瘤干细胞等多种CSC。免疫组化原位染色能有效检测ALDH1活性，从而也可鉴定SC和CSC，目前正在研究将此方法应用于临床评估肿瘤患者的预后。ALDH1已成为分选和鉴定SC和CSC的通用标志物之一，是SC研究的得力工具。
2011, 18(6):686-691.
DOI: 10.3872/j.issn.1007-385X.2011.6.019
Abstract:
在多种恶性肿瘤中，巨噬细胞是浸润到肿瘤中的主要白细胞，被称为肿瘤相关巨噬细胞（tumor-associated macrophage，TAM）。TAM主要来源于血液循环中的单核细胞，属于一类倾向M2型的、分化并不完全的巨噬细胞，具有高度的可塑性。在不同肿瘤，甚至在同一类肿瘤的不同部位、不同生长阶段，TAM膜分子及细胞因子的表达程度不尽相同。TAM通过释放多种细胞因子，促进肿瘤的生长、侵袭、转移、血管和淋巴管发生。调控TAM的募集、分化和信号通路等，是肿瘤免疫治疗的重要方法。
在多种恶性肿瘤中，巨噬细胞是浸润到肿瘤中的主要白细胞，被称为肿瘤相关巨噬细胞（tumor-associated macrophage，TAM）。TAM主要来源于血液循环中的单核细胞，属于一类倾向M2型的、分化并不完全的巨噬细胞，具有高度的可塑性。在不同肿瘤，甚至在同一类肿瘤的不同部位、不同生长阶段，TAM膜分子及细胞因子的表达程度不尽相同。TAM通过释放多种细胞因子，促进肿瘤的生长、侵袭、转移、血管和淋巴管发生。调控TAM的募集、分化和信号通路等，是肿瘤免疫治疗的重要方法。
2011, 18(6):692-695.
DOI: 10.3872/j.issn.1007-385X.2011.6.020
Abstract:
Artemin是胶质细胞源性神经营养因子（gilal cell line derived neurotrophic factor，GDNF）家族中的一个成员，同时也是转化生长因子-β（transforming growth factor-β，TGF-β）超家族中的一个亚型。Artemin通过GDNF受体转导信号调控肿瘤的生长，Artemin参与了胰腺癌、食管癌等消化道肿瘤的发生、发展，并与胰腺癌的嗜神经侵袭密不可分。Artemin可以提高子宫内膜癌、乳腺癌等生殖系统肿瘤细胞的致癌性和侵袭力，降低癌细胞对化疗药物的敏感度。Artemin还在鳞状细胞肺癌和内分泌腺瘤等肿瘤的生长调控中发挥了重要作用，降低Artemin的表达水平可有效抑制肿瘤生长和转移。Artemin有望成为肿瘤诊疗的新靶点。
Artemin是胶质细胞源性神经营养因子（gilal cell line derived neurotrophic factor，GDNF）家族中的一个成员，同时也是转化生长因子-β（transforming growth factor-β，TGF-β）超家族中的一个亚型。Artemin通过GDNF受体转导信号调控肿瘤的生长，Artemin参与了胰腺癌、食管癌等消化道肿瘤的发生、发展，并与胰腺癌的嗜神经侵袭密不可分。Artemin可以提高子宫内膜癌、乳腺癌等生殖系统肿瘤细胞的致癌性和侵袭力，降低癌细胞对化疗药物的敏感度。Artemin还在鳞状细胞肺癌和内分泌腺瘤等肿瘤的生长调控中发挥了重要作用，降低Artemin的表达水平可有效抑制肿瘤生长和转移。Artemin有望成为肿瘤诊疗的新靶点。
2011, 18(6):696-696.
DOI: 10.3872/j.issn.1007-385X.2011.6.021
Abstract:
联系方式
- 《中国肿瘤生物治疗杂志》
- 1994年创刊
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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.