Objective:To study the correlation of nephroblastoma overexpressed gene (NOV) mRNA levels with cell growth- and migration-related gene expression in osteosarcoma tissue. Methods: A total of 78 patients with osteosarco...Objective:To study the correlation of nephroblastoma overexpressed gene (NOV) mRNA levels with cell growth- and migration-related gene expression in osteosarcoma tissue. Methods: A total of 78 patients with osteosarcoma undergoing surgical treatment in our hospital between May 2012 and May 2016 were selected, and the osteosarcoma tissue and paracancerous tissue were collected. Fluorescence quantitative PCR method was used to detect the mRNA expression of NOV as well as pro-proliferation/anti-proliferation genes and invasion genes, and the Pearson test was used to further assess the correlation of NOV mRNA expression with the osteosarcoma cell growth and migration activity.Results: NOV mRNA expression in osteosarcoma tissue was significantly lower than that in paracancerous tissue, FoxM1, STIM1 and Orai1 mRNA expression were significantly higher than those in paracancerous tissue, anti-proliferation genes RanBP9, Tap73, Caspase-3, PTEN and P53 mRNA expression were significantly lower than those in paracancerous tissue, and invasion genes EFEMP1, Notch1, Id1 and Src mRNA expression were significantly higher than those in paracancerous tissue. Pearson test showed that the NOV mRNA expression in osteosarcoma tissue was negatively correlated with pro-proliferation genes FoxM1, STIM1 and Orai1 mRNA expression, positively correlated with anti-proliferation genes RanBP9, Tap73, Caspase-3, PTEN and P53 mRNA expression, and negatively correlated with invasion genes EFEMP1, Notch1, Id1 and Src mRNA expression.Conclusion: NOV expression is relatively low in osteosarcoma tissue, and NOV mRNA expression is negatively correlated with tumor cell proliferation and invasion activity.展开更多
目的:构建能表达靶向肾母细胞瘤过度表达基因(NOV)的小干扰RNA(siRNA)的重组质粒,研究NOV对大鼠肝星状细胞(HSC)活化、增殖、凋亡、分泌细胞外基质(ECM)的影响方法:以NOV为目的基因,以质粒psiRNA- hHlneo为载体,构建能在真核细胞中表达...目的:构建能表达靶向肾母细胞瘤过度表达基因(NOV)的小干扰RNA(siRNA)的重组质粒,研究NOV对大鼠肝星状细胞(HSC)活化、增殖、凋亡、分泌细胞外基质(ECM)的影响方法:以NOV为目的基因,以质粒psiRNA- hHlneo为载体,构建能在真核细胞中表达的靶向NOV的siRNA的重组质粒psiRNA (psiRNA1,2,3)和阴性对照重组质粒pconsiRNA.限制性酶切和测序鉴定构建成功后,脂质体介导重组质粒转染HSC,依据转染质粒的不同将HSC分为psiRNA1组、psiRNA2组、psiRNA3组、阴性对照pconsiRNA组,并以未转染重组质粒的HSC为空白对照组.半定量RT-PCR检测HSC的NOV、α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原、Ⅲ型胶原mRNA表达情况,Western blot检测α-SMA蛋白表达,MTT法检测细胞增殖,流式细胞仪分析细胞凋亡.结果:限制性酶切和测序鉴定表明成功构建了NOVsiRNA表达质粒:与阴性对照组相比,转染外源重组质粒psiRNA2的HSC内NOV、α-SMA的mRNA表达水平下降(73.0% vs 23.2%,51.4% vs 15.1%,均P<0.05),Ⅰ型胶原、Ⅲ型胶原mRNA表达水平明显下降(59.8% vs 17.0%,37.1% vs 6.6%,P<0.05),α-SMA蛋白表达水平下降.与空白对照组相比,转染外源重组质粒psiRNA2的HSC内HSC增殖活性显著降低(24 h:0.172±0.005 vs 0.318±0.018.P<0.05:48 h:0.296±0.004 vs 0.472±0.029,P<0.05:72 h:0.432±0.024 vs 0.672±0.050,P<0.05).psiRNA1组、psiRNA3组的HSC内NOV mRNA、α-SMA、Ⅰ型胶原、Ⅲ型胶原的表达及HSC增殖活性均无明显下降(均P>0.05).结论:NOV可促进HSC增殖、活化及分泌细胞外基质,提示NOV可以作为肝纤维化基因治疗的一个新的靶位点.展开更多
The localization and development of nephroblastoma overexpressed gene (nov) protein-immunoreactive neurons in the brain of E8-P300 rats have been studied using immunocytochemistry and image analysis. Results are as fo...The localization and development of nephroblastoma overexpressed gene (nov) protein-immunoreactive neurons in the brain of E8-P300 rats have been studied using immunocytochemistry and image analysis. Results are as follows: No NOV protein-immunoreactive cells were detected in the rat brain during prenatal development. A few of positive cells were detected at the early postnatal stage. However, the number and the immunoreactivity of these cells increased gradually at later stages. NOV-immunoreactive cells were widely distributed in the rat brain during P30-P60. The number of immunoreactive cells and their intensity also peaked within this stage. The number and staining intensity of NOV-positive cells decreased gradually with age. The positive cells were mainly located in cingulate cortex, striatum, hippocampus, hypothalamus, cerebellum and brain stem. The present results indicate that nov may play an important role in the development and differentiation of brain as well as maintaining the function of展开更多
文摘Objective:To study the correlation of nephroblastoma overexpressed gene (NOV) mRNA levels with cell growth- and migration-related gene expression in osteosarcoma tissue. Methods: A total of 78 patients with osteosarcoma undergoing surgical treatment in our hospital between May 2012 and May 2016 were selected, and the osteosarcoma tissue and paracancerous tissue were collected. Fluorescence quantitative PCR method was used to detect the mRNA expression of NOV as well as pro-proliferation/anti-proliferation genes and invasion genes, and the Pearson test was used to further assess the correlation of NOV mRNA expression with the osteosarcoma cell growth and migration activity.Results: NOV mRNA expression in osteosarcoma tissue was significantly lower than that in paracancerous tissue, FoxM1, STIM1 and Orai1 mRNA expression were significantly higher than those in paracancerous tissue, anti-proliferation genes RanBP9, Tap73, Caspase-3, PTEN and P53 mRNA expression were significantly lower than those in paracancerous tissue, and invasion genes EFEMP1, Notch1, Id1 and Src mRNA expression were significantly higher than those in paracancerous tissue. Pearson test showed that the NOV mRNA expression in osteosarcoma tissue was negatively correlated with pro-proliferation genes FoxM1, STIM1 and Orai1 mRNA expression, positively correlated with anti-proliferation genes RanBP9, Tap73, Caspase-3, PTEN and P53 mRNA expression, and negatively correlated with invasion genes EFEMP1, Notch1, Id1 and Src mRNA expression.Conclusion: NOV expression is relatively low in osteosarcoma tissue, and NOV mRNA expression is negatively correlated with tumor cell proliferation and invasion activity.
文摘目的:构建能表达靶向肾母细胞瘤过度表达基因(NOV)的小干扰RNA(siRNA)的重组质粒,研究NOV对大鼠肝星状细胞(HSC)活化、增殖、凋亡、分泌细胞外基质(ECM)的影响方法:以NOV为目的基因,以质粒psiRNA- hHlneo为载体,构建能在真核细胞中表达的靶向NOV的siRNA的重组质粒psiRNA (psiRNA1,2,3)和阴性对照重组质粒pconsiRNA.限制性酶切和测序鉴定构建成功后,脂质体介导重组质粒转染HSC,依据转染质粒的不同将HSC分为psiRNA1组、psiRNA2组、psiRNA3组、阴性对照pconsiRNA组,并以未转染重组质粒的HSC为空白对照组.半定量RT-PCR检测HSC的NOV、α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原、Ⅲ型胶原mRNA表达情况,Western blot检测α-SMA蛋白表达,MTT法检测细胞增殖,流式细胞仪分析细胞凋亡.结果:限制性酶切和测序鉴定表明成功构建了NOVsiRNA表达质粒:与阴性对照组相比,转染外源重组质粒psiRNA2的HSC内NOV、α-SMA的mRNA表达水平下降(73.0% vs 23.2%,51.4% vs 15.1%,均P<0.05),Ⅰ型胶原、Ⅲ型胶原mRNA表达水平明显下降(59.8% vs 17.0%,37.1% vs 6.6%,P<0.05),α-SMA蛋白表达水平下降.与空白对照组相比,转染外源重组质粒psiRNA2的HSC内HSC增殖活性显著降低(24 h:0.172±0.005 vs 0.318±0.018.P<0.05:48 h:0.296±0.004 vs 0.472±0.029,P<0.05:72 h:0.432±0.024 vs 0.672±0.050,P<0.05).psiRNA1组、psiRNA3组的HSC内NOV mRNA、α-SMA、Ⅰ型胶原、Ⅲ型胶原的表达及HSC增殖活性均无明显下降(均P>0.05).结论:NOV可促进HSC增殖、活化及分泌细胞外基质,提示NOV可以作为肝纤维化基因治疗的一个新的靶位点.
文摘The localization and development of nephroblastoma overexpressed gene (nov) protein-immunoreactive neurons in the brain of E8-P300 rats have been studied using immunocytochemistry and image analysis. Results are as follows: No NOV protein-immunoreactive cells were detected in the rat brain during prenatal development. A few of positive cells were detected at the early postnatal stage. However, the number and the immunoreactivity of these cells increased gradually at later stages. NOV-immunoreactive cells were widely distributed in the rat brain during P30-P60. The number of immunoreactive cells and their intensity also peaked within this stage. The number and staining intensity of NOV-positive cells decreased gradually with age. The positive cells were mainly located in cingulate cortex, striatum, hippocampus, hypothalamus, cerebellum and brain stem. The present results indicate that nov may play an important role in the development and differentiation of brain as well as maintaining the function of
