AIM:To investigate cytokeratin 8(CK8)overexpression during hepatitis C virus(HCV)infection and its pathogenesis,and the effect of ectopic CK8 expression on hepatoma cell lines.METHODS:We successfully established an in...AIM:To investigate cytokeratin 8(CK8)overexpression during hepatitis C virus(HCV)infection and its pathogenesis,and the effect of ectopic CK8 expression on hepatoma cell lines.METHODS:We successfully established an in vitro HCV cell culture system(HCVcc)to investigate the different expression profiles of CK8 in Huh-7-HCV and Huh-7.5-HCV cells.The expression of CK8 at the mRNA level was determined by real-time polymerase chain reaction(RT-PCR).The expression of CK8 at the protein level was evaluated by Western blotting.We then constructed a eukaryotic expression combination vector containing the coding sequence of human full length CK8 gene.CK8cDNA was amplified by reverse transcription-PCR and inserted into pEGFP-C1 and the positive clone pEGFPCK8 was obtained.After confirming the sequence,the recombinant plasmid was transfected into SMMC7721cells with lipofectamine2000 and CK8 expression was detected using inverted fluorescence microscopy,RTPCR and Western blotting.Besides,we identified biological function of CK8 on SMMC7721 cells,including cell proliferation,cell cycle and apoptosis detection.RESULTS:RT-PCR showed that the expression level of CK8 in Huh-7-HCV and Huh-7.5-HCV cells was 2.88and 2.95 times higher than in control cells.Western blot showed that CK8 expression in Huh-7-HCV and Huh-7.5-HCV cells was 2.53 and 3.26 times higher than that in control cells,respectively.We found that CK8at mRNA and protein levels were both significantly increased in HCVcc.CK8 was up-regulated in SMMC7721cells.CK8 expression at the mRNA level was significantly upregulated in SMMC7721/pEGFP-CK8 cells.CK8expression in SMMC7721/pEGFP-CK8 cells was 2.69times higher than in SMMC7721 cells,and was 2.64times higher than in SMMC7721/pEGFP-C1 cells.CK8expression at the protein level in SMMC7721/pEGFPCK8 cells was 2.46 times higher than in SMMC7721cells,and was 2.29 times higher than in SMMC7721/pEGFP-C1 cells.Further analysis demonstrated that forced expression of CK8 slowed cell growth and induced apoptosis of SMMC7721 cells.CONCLUSION:CK8 up-regulation might have a functional role in HCV infection and pathogenesis,and could be a promising target for the treatment of HCV infection.展开更多
目的研究细胞膜角蛋白8(CK8)和乳腺癌耐药蛋白(breast cancer resistant protein,BCRP)作为多靶点治疗逆转耐药的可行性。方法共转染特异的CK8-siRNAs和BCRP-siRNAs至人乳腺癌多药耐药细胞MCF-7/MX,用Western blot方法检测siRNAs对CK8和...目的研究细胞膜角蛋白8(CK8)和乳腺癌耐药蛋白(breast cancer resistant protein,BCRP)作为多靶点治疗逆转耐药的可行性。方法共转染特异的CK8-siRNAs和BCRP-siRNAs至人乳腺癌多药耐药细胞MCF-7/MX,用Western blot方法检测siRNAs对CK8和BCRP蛋白表达的抑制,荧光染色用激光共聚焦显微镜观察细胞膜表面CK8表达量的变化,并用Sulforhodamine B的方法检测转染前后细胞对多种化疗药敏感性的变化。结果MCF-7/MX细胞导入CK8-siRNAs和BCRP-siRNAs后,CK8和BCRP的表达水平均明显降低,且细胞表面CK8染色也明显降低,同时对米托蒽醌等化疗药的敏感性明显提高,耐药表型明显逆转。结论CK8和BCRP在MCF-7/MX耐药细胞中共同高表达且在乳腺癌多药耐药表型的形成中起重要作用,共同抑制CK8和BCRP的表达可以有效逆转乳腺癌的多药耐药,为治疗肿瘤多药耐药提供了一条新的思路。展开更多
目的探讨炎症指标及肿瘤标志物联合检测对肺癌远处转移的预测价值。方法选取河北医科大学第四医院肺癌患者133例作为研究组,根据转移情况分为转移组、未转移组,检测外周血中性粒细胞/淋巴细胞比值(NLR)、细胞因子及肿瘤标志物,筛选肺癌...目的探讨炎症指标及肿瘤标志物联合检测对肺癌远处转移的预测价值。方法选取河北医科大学第四医院肺癌患者133例作为研究组,根据转移情况分为转移组、未转移组,检测外周血中性粒细胞/淋巴细胞比值(NLR)、细胞因子及肿瘤标志物,筛选肺癌远处转移的危险因素,以Logistic回归分析构建模型,采用受试者工作特征(ROC)曲线分析评估该模型对肺癌远处转移的预测效能,并以河北医科大学第一医院98例肺癌患者作为验证组进行验证。结果转移组NLR、白细胞介素(IL)-8、癌胚抗原(CEA)、细胞角蛋白19片段(CYFRA21-1)、神经元特异性烯醇化酶(NSE)水平均明显高于未转移组,差异有统计学意义(P<0.05);多因素Logistic回归分析结果显示,IL-8(OR=1.008,P=0.014)、CYFRA21-1(OR=1.067,P=0.035)水平升高是预测肺癌远处转移的独立危险因素,联合二者建立的风险预测模型为Logit(P)=-2.217+0.008 X IL-8+0.081 X CYFRA21-1,对预测肺癌远处转移有良好的效能,在研究组、验证组中曲线下面积分别为0.820和0.744,灵敏度分别为91.2%和80.0%,准确率分别为78.9%和72.4%。结论IL-8、CYFRA21-1水平升高是肺癌远处转移的独立危险因素,二者联合构建的风险预测模型预测肺癌远处转移具有良好的性能。展开更多
基金Supported by National Natural Science Foundation of China,No.81170393
文摘AIM:To investigate cytokeratin 8(CK8)overexpression during hepatitis C virus(HCV)infection and its pathogenesis,and the effect of ectopic CK8 expression on hepatoma cell lines.METHODS:We successfully established an in vitro HCV cell culture system(HCVcc)to investigate the different expression profiles of CK8 in Huh-7-HCV and Huh-7.5-HCV cells.The expression of CK8 at the mRNA level was determined by real-time polymerase chain reaction(RT-PCR).The expression of CK8 at the protein level was evaluated by Western blotting.We then constructed a eukaryotic expression combination vector containing the coding sequence of human full length CK8 gene.CK8cDNA was amplified by reverse transcription-PCR and inserted into pEGFP-C1 and the positive clone pEGFPCK8 was obtained.After confirming the sequence,the recombinant plasmid was transfected into SMMC7721cells with lipofectamine2000 and CK8 expression was detected using inverted fluorescence microscopy,RTPCR and Western blotting.Besides,we identified biological function of CK8 on SMMC7721 cells,including cell proliferation,cell cycle and apoptosis detection.RESULTS:RT-PCR showed that the expression level of CK8 in Huh-7-HCV and Huh-7.5-HCV cells was 2.88and 2.95 times higher than in control cells.Western blot showed that CK8 expression in Huh-7-HCV and Huh-7.5-HCV cells was 2.53 and 3.26 times higher than that in control cells,respectively.We found that CK8at mRNA and protein levels were both significantly increased in HCVcc.CK8 was up-regulated in SMMC7721cells.CK8 expression at the mRNA level was significantly upregulated in SMMC7721/pEGFP-CK8 cells.CK8expression in SMMC7721/pEGFP-CK8 cells was 2.69times higher than in SMMC7721 cells,and was 2.64times higher than in SMMC7721/pEGFP-C1 cells.CK8expression at the protein level in SMMC7721/pEGFPCK8 cells was 2.46 times higher than in SMMC7721cells,and was 2.29 times higher than in SMMC7721/pEGFP-C1 cells.Further analysis demonstrated that forced expression of CK8 slowed cell growth and induced apoptosis of SMMC7721 cells.CONCLUSION:CK8 up-regulation might have a functional role in HCV infection and pathogenesis,and could be a promising target for the treatment of HCV infection.
文摘目的探讨炎症指标及肿瘤标志物联合检测对肺癌远处转移的预测价值。方法选取河北医科大学第四医院肺癌患者133例作为研究组,根据转移情况分为转移组、未转移组,检测外周血中性粒细胞/淋巴细胞比值(NLR)、细胞因子及肿瘤标志物,筛选肺癌远处转移的危险因素,以Logistic回归分析构建模型,采用受试者工作特征(ROC)曲线分析评估该模型对肺癌远处转移的预测效能,并以河北医科大学第一医院98例肺癌患者作为验证组进行验证。结果转移组NLR、白细胞介素(IL)-8、癌胚抗原(CEA)、细胞角蛋白19片段(CYFRA21-1)、神经元特异性烯醇化酶(NSE)水平均明显高于未转移组,差异有统计学意义(P<0.05);多因素Logistic回归分析结果显示,IL-8(OR=1.008,P=0.014)、CYFRA21-1(OR=1.067,P=0.035)水平升高是预测肺癌远处转移的独立危险因素,联合二者建立的风险预测模型为Logit(P)=-2.217+0.008 X IL-8+0.081 X CYFRA21-1,对预测肺癌远处转移有良好的效能,在研究组、验证组中曲线下面积分别为0.820和0.744,灵敏度分别为91.2%和80.0%,准确率分别为78.9%和72.4%。结论IL-8、CYFRA21-1水平升高是肺癌远处转移的独立危险因素,二者联合构建的风险预测模型预测肺癌远处转移具有良好的性能。
