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Volume 13,Issue 2,2006 Table of Contents
2006, 13(2):79-82.
DOI: 10.3872/j.issn.1007-385X.2006.2.001
Abstract:
2006, 13(2):83-87.
DOI: 10.3872/j.issn.1007-385X.2006.2.002
Abstract:
Objective: To investigate the underlying mechanism of histone deacetylase (HDAC) inhibitor FK228-induced apoptosis of the prostate cancer cell line DU145.Methods: The inhibitory effect of FK228 on DU145 cell growth and its cytotoxicity were determined by MTT assay; cell cycle arrest was detected by flow cytometry assay; morphological change was observed by Giemsa staining; and defined kinase protein levels were determined by Western blot analysis.Results: FK228 obviously inhibited DU145 cells growth, arrested cell cycle at G0/G1 phase, induced cells morphological changes and degraded several kinase proteins, including EGFR, Her2, Raf-1, Src, Cdk4 and IAP member Survivin. The degradation of these kinases blocked Raf-Mek-Erk and PI3K/Akt survival signal pathways, inducing apoptosis. Conclusion: FK228 may induce DU145 cell apoptosis through depletion of multiple kinase proteins and blockade of survival signal pathways of DU145 cells.
Objective: To investigate the underlying mechanism of histone deacetylase (HDAC) inhibitor FK228-induced apoptosis of the prostate cancer cell line DU145.Methods: The inhibitory effect of FK228 on DU145 cell growth and its cytotoxicity were determined by MTT assay; cell cycle arrest was detected by flow cytometry assay; morphological change was observed by Giemsa staining; and defined kinase protein levels were determined by Western blot analysis.Results: FK228 obviously inhibited DU145 cells growth, arrested cell cycle at G0/G1 phase, induced cells morphological changes and degraded several kinase proteins, including EGFR, Her2, Raf-1, Src, Cdk4 and IAP member Survivin. The degradation of these kinases blocked Raf-Mek-Erk and PI3K/Akt survival signal pathways, inducing apoptosis. Conclusion: FK228 may induce DU145 cell apoptosis through depletion of multiple kinase proteins and blockade of survival signal pathways of DU145 cells.
2006, 13(2):88-92.
DOI: 10.3872/j.issn.1007-385X.2006.2.003
Abstract:
Objective: To investigate the inhibitory effects of STAT3 decoy ODNs on the growth of human bladder cancer cell lines in vitro. Methods: STAT3 decoy and scramble ODNs were transfected into the bladder cell lines (T24 cells and 5637 cells) with Sofast, a positive compound transfection agent. MTT assay was used to detect cell growth rate; the transfection efficiency was detected by flow cytometry assay；the locations of FITC labeled decoy ODNs were observed by reflected light fluorescence microscope；and the expression of STAT3 downstream genes, such as bcl-xl, cyclinD1 and dc-myc mRNA and proteins, were examined by RT-PCR and Western bloting. Results: STAT3 decoy ODNs were effectively incorporated into the bladder cancer cells in a dose-dependent manner, and they inhibited the proliferation of cancer cells. The most obvious inhibition rate (40 % to 48%) was found when the concentration of STAT3 decoy ODNs was at 100 nmol/L. The expression levels of Bcl-xl，c-myc and CylinD1 mRNA and protein were also significantly inhibited by 100 nmol/L STAT3 decoy ODNs. Conclusion: STAT3 decoy ODNs can obviously inhibit the proliferation of the bladder cancer cell lines T24 and 5637 , which may be associated with the inhibition of STAT3-mediated gene expression, such as bcl-xl，c-myc, and cylinD1.
Objective: To investigate the inhibitory effects of STAT3 decoy ODNs on the growth of human bladder cancer cell lines in vitro. Methods: STAT3 decoy and scramble ODNs were transfected into the bladder cell lines (T24 cells and 5637 cells) with Sofast, a positive compound transfection agent. MTT assay was used to detect cell growth rate; the transfection efficiency was detected by flow cytometry assay；the locations of FITC labeled decoy ODNs were observed by reflected light fluorescence microscope；and the expression of STAT3 downstream genes, such as bcl-xl, cyclinD1 and dc-myc mRNA and proteins, were examined by RT-PCR and Western bloting. Results: STAT3 decoy ODNs were effectively incorporated into the bladder cancer cells in a dose-dependent manner, and they inhibited the proliferation of cancer cells. The most obvious inhibition rate (40 % to 48%) was found when the concentration of STAT3 decoy ODNs was at 100 nmol/L. The expression levels of Bcl-xl，c-myc and CylinD1 mRNA and protein were also significantly inhibited by 100 nmol/L STAT3 decoy ODNs. Conclusion: STAT3 decoy ODNs can obviously inhibit the proliferation of the bladder cancer cell lines T24 and 5637 , which may be associated with the inhibition of STAT3-mediated gene expression, such as bcl-xl，c-myc, and cylinD1.
XIANG Jin-yi
,
ZHANG Gui-mei
,
GENG Hui
,
YUAN Ye
,
LIU Yi
,
LI Dong
,
XIAO Han
,
WU Feng-hua
,
FENG Zuo-hua
2006, 13(2):93-97.
DOI: 10.3872/j.issn.1007-385X.2006.2.004
Abstract:
Objective: To investigate the inhibitory effect of in vivo non-targeting transfection of recombinant fibronectin polypeptide CH50 against tumors and to study the related mechanisms. Methods: After inoculated with tumor cells, BALB/c mice were injected with CH50 plasmids, control plasmids, and normal saline separately. The growth of the tumor was observed; the expression of genes (such as B7-1, B7-H1 etc.) in tumor tissues was detected by RT-PCR; and the count of T lymphocytes in local tumor tissues was analyzed by flow cytometry. Results: The tumor growth was obviously suppressed by in vivo CH50 expression. The expression of genes (B7-1 and B7-H1) was up-regulated along with the growth of tumor. CH50 increased the ratios of B7-1/B7-H1 and B7-1/B7-DC and suppressed the up-regulation of IL-10 and TGF-β genes. The direct action of CH50 on H22 cells resulted in the down-regulatoin of TGF-β gene. The count of T lymphocytes in tumor tissues of CH50 treatment group was significantly higher than that in other groups. Conclusion: Expression of CH50 by non-targeting transfection can effectively inhibit the growth of tumor; the regulation of the immuno-regulatory genes in tumor microenvironment is an important part of the treatment mechanism of CH50.
Objective: To investigate the inhibitory effect of in vivo non-targeting transfection of recombinant fibronectin polypeptide CH50 against tumors and to study the related mechanisms. Methods: After inoculated with tumor cells, BALB/c mice were injected with CH50 plasmids, control plasmids, and normal saline separately. The growth of the tumor was observed; the expression of genes (such as B7-1, B7-H1 etc.) in tumor tissues was detected by RT-PCR; and the count of T lymphocytes in local tumor tissues was analyzed by flow cytometry. Results: The tumor growth was obviously suppressed by in vivo CH50 expression. The expression of genes (B7-1 and B7-H1) was up-regulated along with the growth of tumor. CH50 increased the ratios of B7-1/B7-H1 and B7-1/B7-DC and suppressed the up-regulation of IL-10 and TGF-β genes. The direct action of CH50 on H22 cells resulted in the down-regulatoin of TGF-β gene. The count of T lymphocytes in tumor tissues of CH50 treatment group was significantly higher than that in other groups. Conclusion: Expression of CH50 by non-targeting transfection can effectively inhibit the growth of tumor; the regulation of the immuno-regulatory genes in tumor microenvironment is an important part of the treatment mechanism of CH50.
LI Yue-min
,
SONG San-tai
,
JIANG Ze-fei
,
XU Jian-ming
,
ZHANG Qi
,
SU Chang-qing
,
QIAN Yan-zhen
,
QIAN Qi-jun
2006, 13(2):98-102.
DOI: 10.3872/j.issn.1007-385X.2006.2.005
Abstract:
Objective: To evaluate the killing effect of tumor-selective replicating adenovirus CNHK300 combined with paclitaxel (chemotherapy agents) on human breast cancer cells over-expressing HER-2. Methods: The telomerase activity in breast cancer cells and MRC-5 cell line were detected semi-quantitively by TRAP-ELISA assay. Virus proliferation assay and cell viability assay were used to evaluate the proliferation and cytolysis selectivity of CNHK300, and the results were compared with those of wtAd5. The breast cancer cells were treated with CNHK300 combining paclitaxel to evaluate the killing effect of the combined regimen. Western-blot was used to detect the expression of adenovirus CNHK300 E1A in cancer and normal cells. Results: The telomerase activity of the 2 breast cancer cell lines were both positive, while telomerase in MRC-5 fibroblast cell was negative. The proliferative ability of CNHK300 virus and its killing effect on breast cancer cells were similar to those of wtAd5. However, CNHK300 exhibited attenuated replicative ability and killing effect in normal fibroblast cell line than wtAd5 did. Combination of CNHK300 and paclitaxel exhibited increased killing effect in breast cancer cells. CNHK300 E1A was expressed in telomerase-positive breast cancer cells but not in the telomerase-negative normal fibroblast cells.
Objective: To evaluate the killing effect of tumor-selective replicating adenovirus CNHK300 combined with paclitaxel (chemotherapy agents) on human breast cancer cells over-expressing HER-2. Methods: The telomerase activity in breast cancer cells and MRC-5 cell line were detected semi-quantitively by TRAP-ELISA assay. Virus proliferation assay and cell viability assay were used to evaluate the proliferation and cytolysis selectivity of CNHK300, and the results were compared with those of wtAd5. The breast cancer cells were treated with CNHK300 combining paclitaxel to evaluate the killing effect of the combined regimen. Western-blot was used to detect the expression of adenovirus CNHK300 E1A in cancer and normal cells. Results: The telomerase activity of the 2 breast cancer cell lines were both positive, while telomerase in MRC-5 fibroblast cell was negative. The proliferative ability of CNHK300 virus and its killing effect on breast cancer cells were similar to those of wtAd5. However, CNHK300 exhibited attenuated replicative ability and killing effect in normal fibroblast cell line than wtAd5 did. Combination of CNHK300 and paclitaxel exhibited increased killing effect in breast cancer cells. CNHK300 E1A was expressed in telomerase-positive breast cancer cells but not in the telomerase-negative normal fibroblast cells.
2006, 13(2):103-106.
DOI: 10.3872/j.issn.1007-385X.2006.2.006
Abstract:
Objective: To investigate the inhibitory effect of RNA interference (RNAi) on the expression of Aurora-A in SKOV3 cells and on proliferation of SKOV3 cells. Methods: Two pairs of oligo small interference RNA (Oligo siRNA) specific for Aurora-A were designed for RNAi and were transferred into SKOV3 cells. The expression of Aurora-A were detected by RT-PCR and Western blot. Furthermore, the cell proliferation and apoptosis were observed by MTT and FCM after transfection. Results:After transfection with Oligo siRNA, mRNA and protein level of Aurora-A gene in SKOV3 cells were obviously reduced, while the inhibitory rate of proliferation and apoptosis rate in SKOV3 cells were increased significantly. Conclusion: The Oligo siRNA specific for Aurora-A can reduce the expression of Aurora-A gene and induce apoptosis of SKOV3 cells.
Objective: To investigate the inhibitory effect of RNA interference (RNAi) on the expression of Aurora-A in SKOV3 cells and on proliferation of SKOV3 cells. Methods: Two pairs of oligo small interference RNA (Oligo siRNA) specific for Aurora-A were designed for RNAi and were transferred into SKOV3 cells. The expression of Aurora-A were detected by RT-PCR and Western blot. Furthermore, the cell proliferation and apoptosis were observed by MTT and FCM after transfection. Results:After transfection with Oligo siRNA, mRNA and protein level of Aurora-A gene in SKOV3 cells were obviously reduced, while the inhibitory rate of proliferation and apoptosis rate in SKOV3 cells were increased significantly. Conclusion: The Oligo siRNA specific for Aurora-A can reduce the expression of Aurora-A gene and induce apoptosis of SKOV3 cells.
CHEN Qiang
,
LI Xiao-feng
,
YE Yun-bin
,
FAN Nan-feng
,
HUANG Wei-wei
,
CHEN Ming-shui
,
LI Jie-yu
,
CHEN Hui-jing
,
CHEN Shu-ping
,
ZHOU Zhi-feng
2006, 13(2):107-111.
DOI: 10.3872/j.issn.1007-385X.2006.2.007
Abstract:
Objective: To observe the anti-tumor effects of transplanting MHC half-matched bone marrow cells combined with spleen cells in a murine model bearing H22 solid tumor. Methods: Female (BALB/c×C57BL/6) F1 mice (CB6F1，H-2Kb/d) were subcutaneously inoculated with H22 cells to develop a solid tumor model; the model mice were taken as recipients. The syngenic, half-matched, and mismatched donor bone marrow cells were from healthy female F1, male C57BL/6(H-2Kb), and male C3H(H-2K) mice, respectively. The tumor weights were measured 18 days after transplantation. The WBC, biochemistry parameters and the formation of chimerism were observed in mice transplanted with MHC half-matched donor cells irradiated by 7.5Gy 60Co. Meanwhile, graft-versus-host disease (GVHD) was assessed and the results were compared between mice transplanted with radiated and non-radiated donor cells. Results: The tumor weights of mice transplanted with donor cells (radiated/non-irradiated by 60Co) were significantly decreased compared with those of the mice receiving chemotherapy alone（P<0.05）. No anti-tumor effect was found to be induced by MHC half-matched cells in mice receiving no pretreatment of chemotherapy. The transplantation of MHC half-matched donor cells radiated by 7.5 Gy 60Co obviously attenuated GVHD and had no adverse effects on peripheral WBC and functions of liver and kidney. Conclusion:Transplantation of 7.5 Gy 60Co irradiated, MHC half-matched bone marrow cells combined with spleen cells can induce a graft-versus-tumor effect in mice bearing H22 tumor and can attenuate GVHD.
Objective: To observe the anti-tumor effects of transplanting MHC half-matched bone marrow cells combined with spleen cells in a murine model bearing H22 solid tumor. Methods: Female (BALB/c×C57BL/6) F1 mice (CB6F1，H-2Kb/d) were subcutaneously inoculated with H22 cells to develop a solid tumor model; the model mice were taken as recipients. The syngenic, half-matched, and mismatched donor bone marrow cells were from healthy female F1, male C57BL/6(H-2Kb), and male C3H(H-2K) mice, respectively. The tumor weights were measured 18 days after transplantation. The WBC, biochemistry parameters and the formation of chimerism were observed in mice transplanted with MHC half-matched donor cells irradiated by 7.5Gy 60Co. Meanwhile, graft-versus-host disease (GVHD) was assessed and the results were compared between mice transplanted with radiated and non-radiated donor cells. Results: The tumor weights of mice transplanted with donor cells (radiated/non-irradiated by 60Co) were significantly decreased compared with those of the mice receiving chemotherapy alone（P<0.05）. No anti-tumor effect was found to be induced by MHC half-matched cells in mice receiving no pretreatment of chemotherapy. The transplantation of MHC half-matched donor cells radiated by 7.5 Gy 60Co obviously attenuated GVHD and had no adverse effects on peripheral WBC and functions of liver and kidney. Conclusion:Transplantation of 7.5 Gy 60Co irradiated, MHC half-matched bone marrow cells combined with spleen cells can induce a graft-versus-tumor effect in mice bearing H22 tumor and can attenuate GVHD.
2006, 13(2):112-115.
DOI: 10.3872/j.issn.1007-385X.2006.2.008
Abstract:
Objective: To establish a recombinant fowlpox virus vector harboring HN gene of Newcastle disease virus and to study its inhibitory effect on in vitro cultured human hepatic carcinoma cell line SMMC7721 and on H22 hepatoma in C57BL/6 mice. Methods: The fowl poxvirus 282 E4 was transfected with HN gene of Newcastle disease virus, and the product (vFVHN) was screened by 5-Bromo-2-deoxyribouridine and identified by RT-PCR and Western blotting. The mortality of SMMC7721 cells was detected by Methylthiazoletetra-zolium staining after vFVHN infection ; the tumor suppression rate of vFVHN' on H22 hepatoma in C57BL/6 mice was detected to study its anti-tumor effect. Results:Fowlpox virus harboring HN gene of Newcastle disease was successfully constructed, and its killing effect on SMMC7721 was 43.37% and its anti-tumor rate on SMMC7721 cells was 20.65%. Conclusion: It is demonstrated that vFVHN can effectively kill SMMC7721 cells cultured in vitro and has inhibitory effect on C57BL/6 mice bearing H22 hepatoma.
Objective: To establish a recombinant fowlpox virus vector harboring HN gene of Newcastle disease virus and to study its inhibitory effect on in vitro cultured human hepatic carcinoma cell line SMMC7721 and on H22 hepatoma in C57BL/6 mice. Methods: The fowl poxvirus 282 E4 was transfected with HN gene of Newcastle disease virus, and the product (vFVHN) was screened by 5-Bromo-2-deoxyribouridine and identified by RT-PCR and Western blotting. The mortality of SMMC7721 cells was detected by Methylthiazoletetra-zolium staining after vFVHN infection ; the tumor suppression rate of vFVHN' on H22 hepatoma in C57BL/6 mice was detected to study its anti-tumor effect. Results:Fowlpox virus harboring HN gene of Newcastle disease was successfully constructed, and its killing effect on SMMC7721 was 43.37% and its anti-tumor rate on SMMC7721 cells was 20.65%. Conclusion: It is demonstrated that vFVHN can effectively kill SMMC7721 cells cultured in vitro and has inhibitory effect on C57BL/6 mice bearing H22 hepatoma.
2006, 13(2):116-119.
DOI: 10.3872/j.issn.1007-385X.2006.2.009
Abstract:
Objective: To explore the mechanism by which dexamethasone inhibits androgen-independent prostate cancer cells. Methods: The effects of dexamethasone and RU486, a blocker of dexamethasone receptor, on DU145 cell growth were abserved. Flow cytometric analysis was performed to examine the effects of dexamethasone on cell cycle. Western blot analysis was employed to examine the effects of dexamethasone on extracellular signal-regulated kinase 1/2 activity and cyclin D1 expression in DU145 cells. Expression of glucocorticoid receptor gene mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Dexamethasone dose dependently inhibited the growth of DU145 cell and caused cell arrest at G0/G1 stage. Western blot indicated that ERK 1/2 activity and cyclin D1 expression were suppressed after 3 d culture with dexamethasone. The effects of dexamethasone were blocked by RU486 when added simultaneously with dexamethasone. RT-PCR showed expression of glucocorticoid receptor mRNA in DU145 cells. Conclusion: Dexamethasone can suppress DU145 cell proliferation and cell cycle, possibly through the decreasing ERK1/2 activity and cyclin D1 expression, indicating glucocorticoid may have potential therapeutic effect on prostate cancer.
Objective: To explore the mechanism by which dexamethasone inhibits androgen-independent prostate cancer cells. Methods: The effects of dexamethasone and RU486, a blocker of dexamethasone receptor, on DU145 cell growth were abserved. Flow cytometric analysis was performed to examine the effects of dexamethasone on cell cycle. Western blot analysis was employed to examine the effects of dexamethasone on extracellular signal-regulated kinase 1/2 activity and cyclin D1 expression in DU145 cells. Expression of glucocorticoid receptor gene mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Dexamethasone dose dependently inhibited the growth of DU145 cell and caused cell arrest at G0/G1 stage. Western blot indicated that ERK 1/2 activity and cyclin D1 expression were suppressed after 3 d culture with dexamethasone. The effects of dexamethasone were blocked by RU486 when added simultaneously with dexamethasone. RT-PCR showed expression of glucocorticoid receptor mRNA in DU145 cells. Conclusion: Dexamethasone can suppress DU145 cell proliferation and cell cycle, possibly through the decreasing ERK1/2 activity and cyclin D1 expression, indicating glucocorticoid may have potential therapeutic effect on prostate cancer.
2006, 13(2):120-123.
DOI: 10.3872/j.issn.1007-385X.2006.2.010
Abstract:
Objective: To explore the therapeutic efficacy and mechanism of herpes simplex virus thymidine kinase (HSV-TK) in blood vessel-targeted treatment of human hepatocellular carcinoma subcutaneously implanted in mice. Methods: Thirty-two BALB/C mice were subcutaneously transplanted with human hepatocellular carcinoma and the tumors were allowed to grow till the volume reached 100 mm3, then the mice were divided into 4 groups: Ganciclovir group （Ⅰ）, Ad group （Ⅱ）, AdCMV-TK+group GCV (Ⅲ) and AdKDR-TK + GCV group (Ⅳ). Recombinant adenovirus or Ad were injected intra-tumorally in all mice. Ganciclovir (GCV) was given at a dose of 100 mg/(kg·d) (ip) on the following day of injection for 10 days. Microvessel density (MVD) of the tumors was determined with the immunohistochemical methods and the growth of the tumors was observed. Results: Compared with groupⅠ, the tumor inhibitory rate was 12.3% in group Ⅲ and 24.5% in group Ⅳ; the inhibition rates were significantly different between group Ⅲ and IV (P<0.05). The mean MVD in group Ⅰ, Ⅱ, Ⅲ and Ⅳ was 37.4±8.6，30.6±7.8，27.6±7.1，and 10.7±4.1(microvessels/mm2) , respectively; significant differences were found between group Ⅲ and Ⅱ(P<0.05)，Ⅳ and Ⅱ(P<0.01)，and Ⅳ and Ⅲ (P<0.01). Conclusion: Our results suggest that AdKDR-TK may effectively restrain the growth of hepatocellular carcinoma through inhibiting angiogenesis.
Objective: To explore the therapeutic efficacy and mechanism of herpes simplex virus thymidine kinase (HSV-TK) in blood vessel-targeted treatment of human hepatocellular carcinoma subcutaneously implanted in mice. Methods: Thirty-two BALB/C mice were subcutaneously transplanted with human hepatocellular carcinoma and the tumors were allowed to grow till the volume reached 100 mm3, then the mice were divided into 4 groups: Ganciclovir group （Ⅰ）, Ad group （Ⅱ）, AdCMV-TK+group GCV (Ⅲ) and AdKDR-TK + GCV group (Ⅳ). Recombinant adenovirus or Ad were injected intra-tumorally in all mice. Ganciclovir (GCV) was given at a dose of 100 mg/(kg·d) (ip) on the following day of injection for 10 days. Microvessel density (MVD) of the tumors was determined with the immunohistochemical methods and the growth of the tumors was observed. Results: Compared with groupⅠ, the tumor inhibitory rate was 12.3% in group Ⅲ and 24.5% in group Ⅳ; the inhibition rates were significantly different between group Ⅲ and IV (P<0.05). The mean MVD in group Ⅰ, Ⅱ, Ⅲ and Ⅳ was 37.4±8.6，30.6±7.8，27.6±7.1，and 10.7±4.1(microvessels/mm2) , respectively; significant differences were found between group Ⅲ and Ⅱ(P<0.05)，Ⅳ and Ⅱ(P<0.01)，and Ⅳ and Ⅲ (P<0.01). Conclusion: Our results suggest that AdKDR-TK may effectively restrain the growth of hepatocellular carcinoma through inhibiting angiogenesis.
2006, 13(2):124-128.
DOI: 10.3872/j.issn.1007-385X.2006.2.011
Abstract:
Objective: To study the inhibitory effect of siRNA on Heparanase (HPA) expression in SKOV3 cells. Methods:Two pairs of 21 bp reverse repeated sequence targeting HPA RNA (spaced by 9 bp nucleotide) were synthesized and were cloned into plasmid Pgenesil-1 to construct recombinant plasmid PGenesil-1(+)-HPA expressing 2 hairpin siRNAs. The inhibition of HPA gene was detected by RT-PCR, real-time PCR and immunohistochemical staining after PGenesil-1(+)-HPA was stably transfected into SKOV3 cells. Results: The recombinant plasmid PGenesil-1(+)-HPA (expressing 2 hairpin siRNAs) was successfully constructed. RT-PCR, real-time PCR and immunohistochemical staining showed that there was a significant decrease in HPA mRNA and protein level in experimental group compared with those in control group. Conclusion: siRNA targeting HPA mRNA can specically suppress the expression of HPA gene in SKOV3 cells; RNA interference method provides a new way for studying the role of HPA and gene therapy of cancer.
Objective: To study the inhibitory effect of siRNA on Heparanase (HPA) expression in SKOV3 cells. Methods:Two pairs of 21 bp reverse repeated sequence targeting HPA RNA (spaced by 9 bp nucleotide) were synthesized and were cloned into plasmid Pgenesil-1 to construct recombinant plasmid PGenesil-1(+)-HPA expressing 2 hairpin siRNAs. The inhibition of HPA gene was detected by RT-PCR, real-time PCR and immunohistochemical staining after PGenesil-1(+)-HPA was stably transfected into SKOV3 cells. Results: The recombinant plasmid PGenesil-1(+)-HPA (expressing 2 hairpin siRNAs) was successfully constructed. RT-PCR, real-time PCR and immunohistochemical staining showed that there was a significant decrease in HPA mRNA and protein level in experimental group compared with those in control group. Conclusion: siRNA targeting HPA mRNA can specically suppress the expression of HPA gene in SKOV3 cells; RNA interference method provides a new way for studying the role of HPA and gene therapy of cancer.
YANG Feng
,
WAN Tao
,
ZHOU Xiang-yang
,
WU Yan-feng
,
LI Nan
,
CHEN Guo-you
,
JIANG Ying-ming
,
CAO Xue-tao
2006, 13(2):129-134.
DOI: 10.3872/j.issn.1007-385X.2006.2.012
Abstract:
Objective： To prepare, purify the recombinant proteins of HSP70-like protein 1(HSP70L1) with a large fragment of chicken ovalbumin (OVA) , and to investigate the bio-function of the fusion protein, providing a basis for further study of the effect and the mechanism of HSP70L1 as an adjuvant. Methods: The vector containing HSP70L1 cDNA and large fragment of OVA was constructed. The expression of OVA-HSP70L1 fusion protein was induced and the products were purified from inclusion bodies by His-Trap metal chelation chromatography and DEAE ion-exchange chromatography. The bio-activity of the fusion protein was examined by detecting its ability to activate dendritic cells and to promote the secretion of cytokines. Results: The vector was successfully constructed and the molecular weight of the fused OVA-HSP70L1 protein (with a purity of over 95%) was 68 000. The fusion protein effectively promoted the maturation of dendritic cells and the production of cytokines such as interleukin-12 and tumor necrosis factor-α. Conclusion: HSP70L1 may be an effective adjuvant in the fusion protein and it may also promote antigen specific Th1 type i mmol/Luno-responses.
Objective： To prepare, purify the recombinant proteins of HSP70-like protein 1(HSP70L1) with a large fragment of chicken ovalbumin (OVA) , and to investigate the bio-function of the fusion protein, providing a basis for further study of the effect and the mechanism of HSP70L1 as an adjuvant. Methods: The vector containing HSP70L1 cDNA and large fragment of OVA was constructed. The expression of OVA-HSP70L1 fusion protein was induced and the products were purified from inclusion bodies by His-Trap metal chelation chromatography and DEAE ion-exchange chromatography. The bio-activity of the fusion protein was examined by detecting its ability to activate dendritic cells and to promote the secretion of cytokines. Results: The vector was successfully constructed and the molecular weight of the fused OVA-HSP70L1 protein (with a purity of over 95%) was 68 000. The fusion protein effectively promoted the maturation of dendritic cells and the production of cytokines such as interleukin-12 and tumor necrosis factor-α. Conclusion: HSP70L1 may be an effective adjuvant in the fusion protein and it may also promote antigen specific Th1 type i mmol/Luno-responses.
2006, 13(2):135-139.
DOI: 10.3872/j.issn.1007-385X.2006.2.013
Abstract:
Objective: To prepare the single-chain antibody of γ-seminoprotein and to determine its biological activity and specific targeting ability. Methods: Rapid translation system was used to express single-chain anti-γ-seminoprotein antibody; immunoprecipitation, Western blotting, immunofluorescent staining and cellular test were used to investigate the activity and the efficiency of the antibody after entering into prostate cancer cells. Results: The single-chain anti-γ-seminoprotein antibody was successfully prepared, which could specifically bind with the prostate cells and could bring magnetic beads into prostate cancer cells within 15 min, and the amount of magnetic beads in prostate cancer cells increased as the culture time prolonged. Control study showed that the antibody conjugated magnetic beads could not enter other normal cells or tumors. Conclusion: The prepared single-chain anti-γ-seminoprotein antibody has satisfactory biological activity and targeting ability, which makes it promising for the early NMR detection and targeting therapy of prostate cancer.
Objective: To prepare the single-chain antibody of γ-seminoprotein and to determine its biological activity and specific targeting ability. Methods: Rapid translation system was used to express single-chain anti-γ-seminoprotein antibody; immunoprecipitation, Western blotting, immunofluorescent staining and cellular test were used to investigate the activity and the efficiency of the antibody after entering into prostate cancer cells. Results: The single-chain anti-γ-seminoprotein antibody was successfully prepared, which could specifically bind with the prostate cells and could bring magnetic beads into prostate cancer cells within 15 min, and the amount of magnetic beads in prostate cancer cells increased as the culture time prolonged. Control study showed that the antibody conjugated magnetic beads could not enter other normal cells or tumors. Conclusion: The prepared single-chain anti-γ-seminoprotein antibody has satisfactory biological activity and targeting ability, which makes it promising for the early NMR detection and targeting therapy of prostate cancer.
2006, 13(2):140-141.
DOI: 10.3872/j.issn.1007-385X.2006.2.014
Abstract:
2006, 13(2):142-144.
DOI: 10.3872/j.issn.1007-385X.2006.2.015
Abstract:
目的： 观察反应停联合VAD方案治疗难治性多发性骨髓瘤的临床疗效。方法： 将16例多发性骨髓瘤患者随机分为两组，治疗组9例采用反应停联合VAD方案，反应停起始剂量为100 mg/d，每周增加50 mg/d,最大剂量为200~400 mg/d，持续12周以上；对照组7例采用单纯VAD方案，28 d为1疗程，治疗3个疗程。结果： 治疗组总有效率为66.7%，对照组总有效率为42.9%，两组间疗效有显著性差异（P＜0.05）。治疗组出现较对照组相对明显的不良反应有便秘、失眠，对症治疗后缓解。结论： 反应停联合VAD方案治疗多发性骨髓瘤疗效明显优于单用化疗组。
目的： 观察反应停联合VAD方案治疗难治性多发性骨髓瘤的临床疗效。方法： 将16例多发性骨髓瘤患者随机分为两组，治疗组9例采用反应停联合VAD方案，反应停起始剂量为100 mg/d，每周增加50 mg/d,最大剂量为200~400 mg/d，持续12周以上；对照组7例采用单纯VAD方案，28 d为1疗程，治疗3个疗程。结果： 治疗组总有效率为66.7%，对照组总有效率为42.9%，两组间疗效有显著性差异（P＜0.05）。治疗组出现较对照组相对明显的不良反应有便秘、失眠，对症治疗后缓解。结论： 反应停联合VAD方案治疗多发性骨髓瘤疗效明显优于单用化疗组。
2006, 13(2):145-146.
DOI: 10.3872/j.issn.1007-385X.2006.2.016
Abstract:
2006, 13(2):147-149.
DOI: 10.3872/j.issn.1007-385X.2006.2.017
Abstract:
含有未甲基化CpG基序的寡核苷酸序列（CpG ODN）能激活宿主的免疫系统，引起先天免疫应答和获得性免疫应答。CpG ODN诱导产生的细胞因子、活化的各种免疫细胞有明显的抗肿瘤作用。CpG ODN可通过多种途径发挥抗肿瘤作用，包括作为单独治疗剂、免疫佐剂或和其他免疫疗法共同治疗，也可以与放疗﹑化疗以及外科手术联合应用治疗肿瘤。CpG ODN的抗肿瘤活性已经在动物模型中得到证明，目前正以CpG ODN的这一特性为基础开发新型的抗肿瘤药物。有几种CpG ODN新药已经进入临床试验阶段，为肿瘤患者的治疗提供新的途径。
含有未甲基化CpG基序的寡核苷酸序列（CpG ODN）能激活宿主的免疫系统，引起先天免疫应答和获得性免疫应答。CpG ODN诱导产生的细胞因子、活化的各种免疫细胞有明显的抗肿瘤作用。CpG ODN可通过多种途径发挥抗肿瘤作用，包括作为单独治疗剂、免疫佐剂或和其他免疫疗法共同治疗，也可以与放疗﹑化疗以及外科手术联合应用治疗肿瘤。CpG ODN的抗肿瘤活性已经在动物模型中得到证明，目前正以CpG ODN的这一特性为基础开发新型的抗肿瘤药物。有几种CpG ODN新药已经进入临床试验阶段，为肿瘤患者的治疗提供新的途径。
2006, 13(2):150-152.
DOI: 10.3872/j.issn.1007-385X.2006.2.018
Abstract:
成体干细胞是一类具有自我更新能力和多向分化潜能的细胞，其“可塑性”的特性在组织和器官损伤修复的临床应用中发挥了重要作用。自体骨髓干细胞移植治疗缺血性心脏病和心肌梗死安全而有效，自体骨骼肌来源的干细胞心肌内注射治疗心肌梗死也有较好疗效；外伤引起的中枢神经系统损伤采用间充质干细胞(MSC)移植治疗，术后症状均有改善；对于眼表面疾病患者来说，自体角膜缘干细胞移植是简单而有效的重建角膜表面和明显提高有效视力的治疗方法；来源于骨髓的成骨祖细胞,在体外生物陶瓷支架上生长扩增后移植于骨损伤部位能显著提高大段骨损伤的治疗效果。有证据显示皮肤和人吸脂术后的脂肪组织内有类似于MSC的多功能干细胞，具有临床应用的良好前景。
成体干细胞是一类具有自我更新能力和多向分化潜能的细胞，其“可塑性”的特性在组织和器官损伤修复的临床应用中发挥了重要作用。自体骨髓干细胞移植治疗缺血性心脏病和心肌梗死安全而有效，自体骨骼肌来源的干细胞心肌内注射治疗心肌梗死也有较好疗效；外伤引起的中枢神经系统损伤采用间充质干细胞(MSC)移植治疗，术后症状均有改善；对于眼表面疾病患者来说，自体角膜缘干细胞移植是简单而有效的重建角膜表面和明显提高有效视力的治疗方法；来源于骨髓的成骨祖细胞,在体外生物陶瓷支架上生长扩增后移植于骨损伤部位能显著提高大段骨损伤的治疗效果。有证据显示皮肤和人吸脂术后的脂肪组织内有类似于MSC的多功能干细胞，具有临床应用的良好前景。
2006, 13(2):153-155.
DOI: 10.3872/j.issn.1007-385X.2006.2.019
Abstract:
Zbtb7基因位于人染色体19p13.3区带，其编码的产物POK家族具有BTB域和Kruppel锌指结构。通过早期小鼠胚胎成纤维细胞（MEFs）的实验，发现POK家族成员之一的Pokemon是ARF特异性转录抑制剂，通过ARF-P53路径对细胞生长进行调节。Pokemon过表达能使小鼠发生淋巴瘤，其还能与癌基因BCL-6联合作用显示原癌基因的活性，促进淋巴瘤的发生。所以认为该基因有望成为重要的肿瘤监视靶点。
Zbtb7基因位于人染色体19p13.3区带，其编码的产物POK家族具有BTB域和Kruppel锌指结构。通过早期小鼠胚胎成纤维细胞（MEFs）的实验，发现POK家族成员之一的Pokemon是ARF特异性转录抑制剂，通过ARF-P53路径对细胞生长进行调节。Pokemon过表达能使小鼠发生淋巴瘤，其还能与癌基因BCL-6联合作用显示原癌基因的活性，促进淋巴瘤的发生。所以认为该基因有望成为重要的肿瘤监视靶点。
2006, 13(2):156-158.
DOI: 10.3872/j.issn.1007-385X.2006.2.020
Abstract:
组蛋白H2A变体H2AX在离其肽链C端4个氨基酸处存在一个高度保守的丝氨酸残基（Ser-139），DNA双链断裂（DSB）一经产生，该残基便能迅速被磷酸化，形成γ-H2AX。γ-H2AX识别抗体是探测DSB存在的金标准；H2AX在细胞周期关卡中起重要作用，H2AX-/-细胞不能正确停止在G2期并进入有丝分裂期；H2AX的丧失不影响V(D)J重排，但有效的类型转换重组需要H2AX；H2AX还在细胞分裂中起重要作用。总之，H2AX是基因组的监视器，DNA损伤一旦发生，H2AX的磷酸化即会被启动，然后招募更多的修复因子共同修复DNA损伤；H2AX在抑制基因组不稳定和癌症放疗疗效预测方面具有一定的作用。
组蛋白H2A变体H2AX在离其肽链C端4个氨基酸处存在一个高度保守的丝氨酸残基（Ser-139），DNA双链断裂（DSB）一经产生，该残基便能迅速被磷酸化，形成γ-H2AX。γ-H2AX识别抗体是探测DSB存在的金标准；H2AX在细胞周期关卡中起重要作用，H2AX-/-细胞不能正确停止在G2期并进入有丝分裂期；H2AX的丧失不影响V(D)J重排，但有效的类型转换重组需要H2AX；H2AX还在细胞分裂中起重要作用。总之，H2AX是基因组的监视器，DNA损伤一旦发生，H2AX的磷酸化即会被启动，然后招募更多的修复因子共同修复DNA损伤；H2AX在抑制基因组不稳定和癌症放疗疗效预测方面具有一定的作用。
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.