AIM: To assess human cytomegalovirus-encoded US28 gene function in colorectal cancer(CRC) pathogenesis.METHODS: Immunohistochemical analysis was performed to determine US28 expression in 103 CRC patient samples and 98...AIM: To assess human cytomegalovirus-encoded US28 gene function in colorectal cancer(CRC) pathogenesis.METHODS: Immunohistochemical analysis was performed to determine US28 expression in 103 CRC patient samples and 98 corresponding adjacent noncancerous samples. Patient data were compared by age, sex, tumor location, histological grade, Dukes' stage, and overall mean survival time. In addition, the US28 gene was transiently transfected into the CRC LOVO cell line, and cell proliferation was assessed using a cell counting kit-8 assay. Cell cycle analysis by flow cytometry and a cell invasion transwell assay were also carried out.RESULTS: US28 levels were clearly higher in CRC tissues(38.8%) than in adjacent noncancerous samples(7.1%)(P = 0.000). Interestingly, elevated US28 amounts in CRC tissues were significantly associated with histological grade, metastasis, Dukes' stage, and overall survival(all P < 0.05); meanwhile, US28 expression was not significantly correlated with age, sex or tumor location. In addition, multivariate Coxregression data revealed US28 level as an independent CRC prognostic marker(P = 0.000). LOVO cells successfully transfected with the US28 gene exhibited higher viability, greater chemotherapy resistance, accelerated cell cycle progression, and increased invasion ability.CONCLUSION: US28 expression is predictive of poor prognosis and may promote CRC.展开更多
目的利用酵母双杂交技术筛选与人巨细胞病毒US28蛋白相互作用的宿主蛋白,可为研究US28蛋白的作用机制提供依据。方法构建p GBKT7-US28诱饵重组载体,检测其在酵母细胞中的表达和自激活作用,然后利用酵母双杂交系统筛选人脑文库中与US28...目的利用酵母双杂交技术筛选与人巨细胞病毒US28蛋白相互作用的宿主蛋白,可为研究US28蛋白的作用机制提供依据。方法构建p GBKT7-US28诱饵重组载体,检测其在酵母细胞中的表达和自激活作用,然后利用酵母双杂交系统筛选人脑文库中与US28相互作用的蛋白质。对获得的阳性克隆进行PCR鉴定、测序及序列比对分析,并通过酵母共转化实验再次确认蛋白的相互作用。结果 p GBKT7-US28在酵母中成功表达US28融合蛋白;酵母双杂交筛选并验证,获得了5个与US28蛋白相互作用的宿主蛋白。结论初步鉴定得到5个与US28相互作用的细胞蛋白,为探索US28蛋白的新功能以及揭示巨细胞病毒的致病机理和疾病治疗奠定基础。展开更多
基金Supported by The Zhejiang Provincial Natural Science Foundation of China,No.LY15H160059Zhejiang Provincial Medical and Health Science and Technology Project,No.2016KYB192the Wenzhou Municipal Science and Technology Bureau,No.Y20140691
文摘AIM: To assess human cytomegalovirus-encoded US28 gene function in colorectal cancer(CRC) pathogenesis.METHODS: Immunohistochemical analysis was performed to determine US28 expression in 103 CRC patient samples and 98 corresponding adjacent noncancerous samples. Patient data were compared by age, sex, tumor location, histological grade, Dukes' stage, and overall mean survival time. In addition, the US28 gene was transiently transfected into the CRC LOVO cell line, and cell proliferation was assessed using a cell counting kit-8 assay. Cell cycle analysis by flow cytometry and a cell invasion transwell assay were also carried out.RESULTS: US28 levels were clearly higher in CRC tissues(38.8%) than in adjacent noncancerous samples(7.1%)(P = 0.000). Interestingly, elevated US28 amounts in CRC tissues were significantly associated with histological grade, metastasis, Dukes' stage, and overall survival(all P < 0.05); meanwhile, US28 expression was not significantly correlated with age, sex or tumor location. In addition, multivariate Coxregression data revealed US28 level as an independent CRC prognostic marker(P = 0.000). LOVO cells successfully transfected with the US28 gene exhibited higher viability, greater chemotherapy resistance, accelerated cell cycle progression, and increased invasion ability.CONCLUSION: US28 expression is predictive of poor prognosis and may promote CRC.
文摘目的利用酵母双杂交技术筛选与人巨细胞病毒US28蛋白相互作用的宿主蛋白,可为研究US28蛋白的作用机制提供依据。方法构建p GBKT7-US28诱饵重组载体,检测其在酵母细胞中的表达和自激活作用,然后利用酵母双杂交系统筛选人脑文库中与US28相互作用的蛋白质。对获得的阳性克隆进行PCR鉴定、测序及序列比对分析,并通过酵母共转化实验再次确认蛋白的相互作用。结果 p GBKT7-US28在酵母中成功表达US28融合蛋白;酵母双杂交筛选并验证,获得了5个与US28蛋白相互作用的宿主蛋白。结论初步鉴定得到5个与US28相互作用的细胞蛋白,为探索US28蛋白的新功能以及揭示巨细胞病毒的致病机理和疾病治疗奠定基础。
