目的研究黄芪多糖(APS)对人外周血单核细胞(PBMC)源性树突状细胞(DCs)的基因表达和其功能变化的影响,进一步探讨黄芪多糖的抗AS的作用机制。方法以健康人外周血分离PBMC源性DCs和血清作为研究对象,培育5 d后,随机分为APS组和对照组。其...目的研究黄芪多糖(APS)对人外周血单核细胞(PBMC)源性树突状细胞(DCs)的基因表达和其功能变化的影响,进一步探讨黄芪多糖的抗AS的作用机制。方法以健康人外周血分离PBMC源性DCs和血清作为研究对象,培育5 d后,随机分为APS组和对照组。其中APS组给予200 mg/L APS孵育过夜,对照组不给干预。通过基因芯片和RT-PCR技术,观察APS处理PBMC源性DCs的免疫功能和其他基因表达的差异与AS发生发展的关系。结果与对照组相比,APS组CD36(0.97±0.23 vs5.45±1.14)、IL-27(1.08±0.22 vs 2.97±0.61)基因表达相对量显著性上调;APS组IFI16(0.98±0.18 vs 0.46±0.11)基因表达相对量显著性下调。结论适量的APS可以上调DCs膜表面与抗原递呈相关的CD36、IL-27的表达,下调IFI16的表达,对增加DCs的免疫活性,促进DCs的成熟与分化有显著的影响。APS对AS的发生发展具有明显的积极地干预作用,有着重要的积极地临床意义。展开更多
AIM: To investigate the killing effect and radiosensitization of double suicide gene mediated by adenovirus on colorectal carcinoma cells. METHODS: Colorectal carcinoma cell line SW480 was transfected with adenovirus ...AIM: To investigate the killing effect and radiosensitization of double suicide gene mediated by adenovirus on colorectal carcinoma cells. METHODS: Colorectal carcinoma cell line SW480 was transfected with adenovirus expression vector containing cytosine deaminase (CD) and thymidine kinase (TK) fusion gene. The expression of CD-TK fusion gene was detected by reverse transcriptase-polymerase chain reaction. The toxic effect of ganciclovir (GCV) and 5-fluorocytosine (5-FC) on infected cells was determined by MTT assay. The radiosensitization of double suicide gene was evaluated by clonogenic assay. RESULTS: After prodrugs were used, the survival rate of colorectal carcinoma cells was markedly decreased. When GCV and 5-FC were used in combination, the cytotoxicity and bystander effect were markedly superior to a single prodrug (X2 = 30.371, P<0.01). Both GCV and 5-FC could sensitize colorectal carcinoma cells to the toxic effect of radiation, and greater radiosensitization was achieved when both prodrug were used in combination. CONCLUSION: CD-TK double suicide gene can kill and radiosensitize colorectal carcinoma cells.展开更多
INTRODUCTIONFas/ FasL system has been identified as a keymediator of apoptosis in tumor cells[1-4]. Theoccurrence and development of neoplasm are closelyrelated to apoptosis[5-7] Most chemotherapeuticdrugs kill cancer...INTRODUCTIONFas/ FasL system has been identified as a keymediator of apoptosis in tumor cells[1-4]. Theoccurrence and development of neoplasm are closelyrelated to apoptosis[5-7] Most chemotherapeuticdrugs kill cancer cells mainly by inducingapoptosis[8-14].'展开更多
CD59, belonging to membrane complement regulatory proteins （mCRPs）, inhibits the cytolytic activity of complement and is over-expressed in solid cancers, including ovary cancer. The aim of the present study was to c...CD59, belonging to membrane complement regulatory proteins （mCRPs）, inhibits the cytolytic activity of complement and is over-expressed in solid cancers, including ovary cancer. The aim of the present study was to construct recombinant retrovirus encoding shRNA targeted human CD59 and infect A2780 cells in order to investigate the relationship between decreased CD59 expression and tumorigenesis of ovary cancer, siCD59 and siCD59-C were successfully constructed and identified by PCR, restriction endonuclease analyses and DNA sequencing, respectively. The siCD59 was able to efficiently infect A2780 cells, which was confirmed by Western blotting. When incubated with fresh normal human serum （8%, v/v） for 1 h at 37℃, the cell viability was decreased and cell damage was increased in siCD59 infected A2780 cells compared to siCD59-C infected cells. This led to the activation of caspase-3. The apoptosis in siCD59 infected cells was shown with hypercondensed nuclei using Hoechst staining. Meanwhile, the weight of ovary tumor graft in nude mice was significantly decreased in siCD59 group compared to that of siCD59-C group. And the expression of CD59 protein in tumor tissue in siCD59 group was significantly decreased. These results suggested that CD59 silencing in ovary cancer cells v/a retrovirusmediated RNAi can enhance complement-mediated cell damage, inhibiting growth of ovary cancer. CD59 might be a potential target for gene therapy in ovary cancer. Cellular ＆ Molecular Immunology.展开更多
文摘目的研究黄芪多糖(APS)对人外周血单核细胞(PBMC)源性树突状细胞(DCs)的基因表达和其功能变化的影响,进一步探讨黄芪多糖的抗AS的作用机制。方法以健康人外周血分离PBMC源性DCs和血清作为研究对象,培育5 d后,随机分为APS组和对照组。其中APS组给予200 mg/L APS孵育过夜,对照组不给干预。通过基因芯片和RT-PCR技术,观察APS处理PBMC源性DCs的免疫功能和其他基因表达的差异与AS发生发展的关系。结果与对照组相比,APS组CD36(0.97±0.23 vs5.45±1.14)、IL-27(1.08±0.22 vs 2.97±0.61)基因表达相对量显著性上调;APS组IFI16(0.98±0.18 vs 0.46±0.11)基因表达相对量显著性下调。结论适量的APS可以上调DCs膜表面与抗原递呈相关的CD36、IL-27的表达,下调IFI16的表达,对增加DCs的免疫活性,促进DCs的成熟与分化有显著的影响。APS对AS的发生发展具有明显的积极地干预作用,有着重要的积极地临床意义。
文摘AIM: To investigate the killing effect and radiosensitization of double suicide gene mediated by adenovirus on colorectal carcinoma cells. METHODS: Colorectal carcinoma cell line SW480 was transfected with adenovirus expression vector containing cytosine deaminase (CD) and thymidine kinase (TK) fusion gene. The expression of CD-TK fusion gene was detected by reverse transcriptase-polymerase chain reaction. The toxic effect of ganciclovir (GCV) and 5-fluorocytosine (5-FC) on infected cells was determined by MTT assay. The radiosensitization of double suicide gene was evaluated by clonogenic assay. RESULTS: After prodrugs were used, the survival rate of colorectal carcinoma cells was markedly decreased. When GCV and 5-FC were used in combination, the cytotoxicity and bystander effect were markedly superior to a single prodrug (X2 = 30.371, P<0.01). Both GCV and 5-FC could sensitize colorectal carcinoma cells to the toxic effect of radiation, and greater radiosensitization was achieved when both prodrug were used in combination. CONCLUSION: CD-TK double suicide gene can kill and radiosensitize colorectal carcinoma cells.
文摘INTRODUCTIONFas/ FasL system has been identified as a keymediator of apoptosis in tumor cells[1-4]. Theoccurrence and development of neoplasm are closelyrelated to apoptosis[5-7] Most chemotherapeuticdrugs kill cancer cells mainly by inducingapoptosis[8-14].'
基金supported by grants from the National Natural Science Foundation of China (No. 30671936).
文摘CD59, belonging to membrane complement regulatory proteins （mCRPs）, inhibits the cytolytic activity of complement and is over-expressed in solid cancers, including ovary cancer. The aim of the present study was to construct recombinant retrovirus encoding shRNA targeted human CD59 and infect A2780 cells in order to investigate the relationship between decreased CD59 expression and tumorigenesis of ovary cancer, siCD59 and siCD59-C were successfully constructed and identified by PCR, restriction endonuclease analyses and DNA sequencing, respectively. The siCD59 was able to efficiently infect A2780 cells, which was confirmed by Western blotting. When incubated with fresh normal human serum （8%, v/v） for 1 h at 37℃, the cell viability was decreased and cell damage was increased in siCD59 infected A2780 cells compared to siCD59-C infected cells. This led to the activation of caspase-3. The apoptosis in siCD59 infected cells was shown with hypercondensed nuclei using Hoechst staining. Meanwhile, the weight of ovary tumor graft in nude mice was significantly decreased in siCD59 group compared to that of siCD59-C group. And the expression of CD59 protein in tumor tissue in siCD59 group was significantly decreased. These results suggested that CD59 silencing in ovary cancer cells v/a retrovirusmediated RNAi can enhance complement-mediated cell damage, inhibiting growth of ovary cancer. CD59 might be a potential target for gene therapy in ovary cancer. Cellular ＆ Molecular Immunology.
