CUEDC2(CUE domain containging protein 2)是近年来发现的一个功能尚未十分明确的蛋白质。实验室前期的研究表明,CUEDC2通过影响孕激素受体PR抑制乳腺癌细胞生长。此外,研究还发现CUEDC2通过招募PP1磷酸酶,促进IKK复合体的去磷酸化,抑...CUEDC2(CUE domain containging protein 2)是近年来发现的一个功能尚未十分明确的蛋白质。实验室前期的研究表明,CUEDC2通过影响孕激素受体PR抑制乳腺癌细胞生长。此外,研究还发现CUEDC2通过招募PP1磷酸酶,促进IKK复合体的去磷酸化,抑制NF-kB信号通路的激活。为了深入研究CUEDC2的功能,构建了CUED2可诱导表达载体(p617-neo-T-CUEDC2-I-tTA4),并通过逆转录病毒系统获得tet-off可诱导表达CUEDC2的稳定细胞系。该诱导表达细胞株的建立为CUEDC2功能基因的研究提供了必要的手段。展开更多
The basal activity of JNK is low in normal growing cells and inactivated JNK targets p53 for ubiquitination. To elucidate if the C-terminal part of JNK is responsible for its binding to p53, the low background tet-off...The basal activity of JNK is low in normal growing cells and inactivated JNK targets p53 for ubiquitination. To elucidate if the C-terminal part of JNK is responsible for its binding to p53, the low background tet-off inducible NIH3T3 cell line was selected by luciferase reporter gene and a double stable C-JNK Aa (203-424) cell line was established. After withdrawing tetracycline, the C-JNK fragment expression was induced and cell growth was dramati- cally inhibited 24 h later. However, the expresion of p53 was found to be increased after the induction of C-JNK fragment, evaluated by transfecting p21waf-luciferase reporter genes. Our further studies showed that C-JNK fragment could form complex with p53 both in vivo and in vitro. Induction of C-JNK fragment in vivo can increase p53 stability by inhibiting p53 ubiquitination.展开更多
Liver cancer is one of the most threatening diseases in Chinese population. Just like in other tissues, tumor initiation and development in liver involve multiple steps of genetic and epigenetic alterations with sever...Liver cancer is one of the most threatening diseases in Chinese population. Just like in other tissues, tumor initiation and development in liver involve multiple steps of genetic and epigenetic alterations with several unknown details. However, unlike in other tissues, a tissue specific inducible Cre recombinase system that allows temporal and spatial deletion of a target DNA fragment is still not available for in vivo functional gene annotation in hepatocytes. In our pursuit to establish such a mouse model, we designed a dual inducible Cre transgene system and tested it in cultured cells. By combining a CCAAT/enhancer binding protein β （C/EBP β） promoter derived Tet-off expression system and the estrogen receptor （ER） mediated functional control, we show a desirable profile of both hepatocyte-specificity and regulability of the Cre expression in a series of critical assessments in the cell culture system, which provides confidence in continuation of our ongoing pursuit in mouse.展开更多
文摘CUEDC2(CUE domain containging protein 2)是近年来发现的一个功能尚未十分明确的蛋白质。实验室前期的研究表明,CUEDC2通过影响孕激素受体PR抑制乳腺癌细胞生长。此外,研究还发现CUEDC2通过招募PP1磷酸酶,促进IKK复合体的去磷酸化,抑制NF-kB信号通路的激活。为了深入研究CUEDC2的功能,构建了CUED2可诱导表达载体(p617-neo-T-CUEDC2-I-tTA4),并通过逆转录病毒系统获得tet-off可诱导表达CUEDC2的稳定细胞系。该诱导表达细胞株的建立为CUEDC2功能基因的研究提供了必要的手段。
基金supported by National Natural Science Foundation of China(No.30270556)The National Basic Research Program(No.2002CB513004).
文摘The basal activity of JNK is low in normal growing cells and inactivated JNK targets p53 for ubiquitination. To elucidate if the C-terminal part of JNK is responsible for its binding to p53, the low background tet-off inducible NIH3T3 cell line was selected by luciferase reporter gene and a double stable C-JNK Aa (203-424) cell line was established. After withdrawing tetracycline, the C-JNK fragment expression was induced and cell growth was dramati- cally inhibited 24 h later. However, the expresion of p53 was found to be increased after the induction of C-JNK fragment, evaluated by transfecting p21waf-luciferase reporter genes. Our further studies showed that C-JNK fragment could form complex with p53 both in vivo and in vitro. Induction of C-JNK fragment in vivo can increase p53 stability by inhibiting p53 ubiquitination.
基金supported by Shanghai Science Foundation grants,National Science Foundation of China(No.30570850 and 10574134)National Research Program for Basic Research of China(No.2004CB518804)+1 种基金National Research Program for High Technology(No.2006AA02Z-320 and 2006AA 02Z197)European 6th Program(LSHBCT-2005-019067).
文摘Liver cancer is one of the most threatening diseases in Chinese population. Just like in other tissues, tumor initiation and development in liver involve multiple steps of genetic and epigenetic alterations with several unknown details. However, unlike in other tissues, a tissue specific inducible Cre recombinase system that allows temporal and spatial deletion of a target DNA fragment is still not available for in vivo functional gene annotation in hepatocytes. In our pursuit to establish such a mouse model, we designed a dual inducible Cre transgene system and tested it in cultured cells. By combining a CCAAT/enhancer binding protein β （C/EBP β） promoter derived Tet-off expression system and the estrogen receptor （ER） mediated functional control, we show a desirable profile of both hepatocyte-specificity and regulability of the Cre expression in a series of critical assessments in the cell culture system, which provides confidence in continuation of our ongoing pursuit in mouse.
