论文部分内容阅读
In order to provide the experimental basis for the further studies on the oncogenic mechanism of the E6 protein from human papillomavirus type 16(HPV16),the eukaryotie expression vector pcDNA3.1(-)/E6 was used for the study on the effect of E6 protein to influence the secretory activity of LPS-indueed THP-1-macrophages,and the reconstructed plasmid pcDNA3.1(-)/E6 was transfected into THP-1-maerophages.The expression of E6 gene was assayed in macrophage lysates by using Western blot analysis and the level of TNF-αor IL-1βwas examined by ELISA.All of data were analyzed by SPSS12.0.As demonstrated by Western blot analysis,the expression of E6 protein with a molecular weight of about 18 kDa by plasmid pcDNA3.1(-)/E6 in THP-1-macrophages could be detected.Howev- er,as demonstrated by ELISA assay,the level of TNF-αor IL-1βin lysates of THP-1-macrophages showed an obvious difference between the pcDNA3.1(-)/E6 group and the LPS control group or the pcDNA3.1(-)control group(P<0.01),but no significant difference existed between pcDNA3.1(-) control group and LPS control group(P>0.05).All these results illustrate that the transient over-ex- pression of HPV6 E6 protein reduces the production of TNF-αand IL-1βinduced by LPS in THP-1-mac- rophages.
In order to provide the experimental basis for the further studies on the oncogenic mechanism of the E6 protein from human papillomavirus type 16 (HPV16), the eukaryotie expression vector pcDNA3.1 (-) / E6 was used for the study on the effect of E6 protein to influence the secretory activity of LPS-indueed THP-1-macrophages, and the reconstructed plasmid pcDNA3.1 (-) / E6 was transfected into THP-1-maerophages. The expression of E6 gene was assayed in macrophage lysates by using Western blot analysis and the level of TNF-αor IL-1βwas examined by ELISA. All of the data were analyzed by SPSS12.0. As demonstrated by Western blot analysis, the expression of E6 protein with a molecular weight of about 18 kDa by plasmid pcDNA3. 1 (-) / E6 in THP-1-macrophages could be detected. Host-er, as demonstrated by ELISA assay, the level of TNF-αor IL-1βin lysates of THP-1-macrophages showed an obvious difference between the pcDNA3. 1 (-) / E6 group and the LPS control group or the pcDNA3.1 (-) control group (P <0.01), but no s ignificant difference originally between pcDNA3.1 (-) control group and LPS control group (P> 0.05). All these results illustrate that the transient over-ex pression of HPV6 E6 protein reduces the production of TNF-α and IL-1 β induced by LPS in THP-1-mac-rophages.