As a core member of the Hippo signaling pathway,Hpo plays a critical role in regulating growth and development.Previous studies reported that loss of function of Hpo results in increased proliferation,reduced apoptosi...As a core member of the Hippo signaling pathway,Hpo plays a critical role in regulating growth and development.Previous studies reported that loss of function of Hpo results in increased proliferation,reduced apoptosis and induction of tissue overgrowth in Drosophila.In this study,we used CRISPR/Cas9(Clustered Regularly Interspaced Short Palindromic Repeats/Casg)to study Hpo gene (BmHpo)function in the lepidopteran insect Bombyx mori,known commonly as the silkworm.Sequence analysis of BmHpo revealed an array of deletions in mutants.We found that BmHpo knockout resulted in defects in body size regnlation,in developmental defects and pigment accumulation and early death. Our data show that BmHpo is essential for regulation of insect growth and development and that CRISPR/Cas9technology can serve as a basis for functional analysis of target genes in lepidopteran insects.展开更多
Signal transduction plays important roles in biological systems. Unfortunately, our knowledge about signaling pathways is far from complete. Specifically, the direction of signaling flows is less known even though the...Signal transduction plays important roles in biological systems. Unfortunately, our knowledge about signaling pathways is far from complete. Specifically, the direction of signaling flows is less known even though the signaling molecules of some signaling pathways have been determined. In this paper, we propose a novel hybrid intelligent method, namely HISP （Hybrid Intelligent approach for identifying directed Signaling Pathways）, to determine both the topologies of signaling pathways and the direction of signaling flows within a pathway based on integer linear programming and genetic algorithm. By integrating the protein-protein interaction, gene expression, and gene knockout data, our HISP approach is able to determine the optimal topologies of signaling pathways in an accurate way. Benchmark results on yeast MAPK signaling pathways demonstrate the efficiency of our proposed approach. When applied to the EGFR/ErbB signaling pathway in human hepatocytes, HISP unveils a high-resolution signaling path- way, where many signaling interactions were missing by existing computational approaches.展开更多
Cells in plants, like in animals, constantly communicate with one another to coordinate their cellular activities in response to surrounding environmental conditions. Higher plants are multicellular organisms. A plant...Cells in plants, like in animals, constantly communicate with one another to coordinate their cellular activities in response to surrounding environmental conditions. Higher plants are multicellular organisms. A plant is originated from a single fertilized egg, which eventually develops into an intact individual with all necessary tissues and organs. Cell-to-cell communication is therefore especially critical throughout its life span, from embryo development, to tissue and organ formation, and finally to flowering and senescence, in addition.展开更多
Objective This study investigated the role of the STAT3/survivin signaling pathway in the EML4-ALK- positive lung adenocarcinoma cell line H2228 before and after crizotinib-induced resistance. The mecha- nism of resis...Objective This study investigated the role of the STAT3/survivin signaling pathway in the EML4-ALK- positive lung adenocarcinoma cell line H2228 before and after crizotinib-induced resistance. The mecha- nism of resistance was studied. Methods Cell viability was determined using the MTT assay. Crizotinib-induced apoptosis in H2228 and H2228 crizotinib-resistant cells treated with the indicated doses of crizotinib was measured at different times （24 h, 48 h, 72 h） using flow cytometry. The levels of p-ALK, ALK, p-STAT3, STAT3, and survivin after treatment of cells with 0, 0.3, and 1 pM crizotinib for 72 h were determined using Western blot analysis. DNA sequencing was used to identify mutations in H2228 crizotinib-resistant cells. Results The crizotinib IC50 values in H2228 and H2228 crizotinib-resistant cells at 72 h were 334.5 nM and 3418 nM, respectively. The resistance index of 1-12228 crizotinib-resistant cells was 10.20. Crizotinib induced apoptosis in H2228 cells and reduced the levels of p-ALK, p-STAT3, and survivin. In contrast, no changes in the levels of p-ALK, p-STAT3, and survivin were observed in H2228 crizotinib-resistant cells. The mutations 2067G--,A and 2182G--,C in EML4-ALK were present in the H2228 crizotinib-resistant cells. Conclusion Crizotinib decreased the viability of H2228 cells in a dose- and time-dependent manner. In the STAT3/survivin pathway, downregulation of p-ALK, p-STAT3, and survivin might contribute to crizo- tinib-induced apoptosis in H2228 ceils. However, the STAT3/survivin pathway in H2228 crizotinib-resistant cells was unaffected by crizotinib treatment. Acquired resistance in H2228 cells might be related to ALK mutations.展开更多
基金grants from the National Natural Science Foundation of China(31772517 and 31530072) Science and Technology Commission of Shanghai Municipality(No.18140902200).
文摘As a core member of the Hippo signaling pathway,Hpo plays a critical role in regulating growth and development.Previous studies reported that loss of function of Hpo results in increased proliferation,reduced apoptosis and induction of tissue overgrowth in Drosophila.In this study,we used CRISPR/Cas9(Clustered Regularly Interspaced Short Palindromic Repeats/Casg)to study Hpo gene (BmHpo)function in the lepidopteran insect Bombyx mori,known commonly as the silkworm.Sequence analysis of BmHpo revealed an array of deletions in mutants.We found that BmHpo knockout resulted in defects in body size regnlation,in developmental defects and pigment accumulation and early death. Our data show that BmHpo is essential for regulation of insect growth and development and that CRISPR/Cas9technology can serve as a basis for functional analysis of target genes in lepidopteran insects.
文摘Signal transduction plays important roles in biological systems. Unfortunately, our knowledge about signaling pathways is far from complete. Specifically, the direction of signaling flows is less known even though the signaling molecules of some signaling pathways have been determined. In this paper, we propose a novel hybrid intelligent method, namely HISP （Hybrid Intelligent approach for identifying directed Signaling Pathways）, to determine both the topologies of signaling pathways and the direction of signaling flows within a pathway based on integer linear programming and genetic algorithm. By integrating the protein-protein interaction, gene expression, and gene knockout data, our HISP approach is able to determine the optimal topologies of signaling pathways in an accurate way. Benchmark results on yeast MAPK signaling pathways demonstrate the efficiency of our proposed approach. When applied to the EGFR/ErbB signaling pathway in human hepatocytes, HISP unveils a high-resolution signaling path- way, where many signaling interactions were missing by existing computational approaches.
文摘Cells in plants, like in animals, constantly communicate with one another to coordinate their cellular activities in response to surrounding environmental conditions. Higher plants are multicellular organisms. A plant is originated from a single fertilized egg, which eventually develops into an intact individual with all necessary tissues and organs. Cell-to-cell communication is therefore especially critical throughout its life span, from embryo development, to tissue and organ formation, and finally to flowering and senescence, in addition.
基金Supported by grants from the Bureau of Science and Technology,Guangxi Zhuang Autonomous Zone,China(No.201017)National Natural Science Foundation of China(No.81060188 and 81260357)
文摘Objective This study investigated the role of the STAT3/survivin signaling pathway in the EML4-ALK- positive lung adenocarcinoma cell line H2228 before and after crizotinib-induced resistance. The mecha- nism of resistance was studied. Methods Cell viability was determined using the MTT assay. Crizotinib-induced apoptosis in H2228 and H2228 crizotinib-resistant cells treated with the indicated doses of crizotinib was measured at different times （24 h, 48 h, 72 h） using flow cytometry. The levels of p-ALK, ALK, p-STAT3, STAT3, and survivin after treatment of cells with 0, 0.3, and 1 pM crizotinib for 72 h were determined using Western blot analysis. DNA sequencing was used to identify mutations in H2228 crizotinib-resistant cells. Results The crizotinib IC50 values in H2228 and H2228 crizotinib-resistant cells at 72 h were 334.5 nM and 3418 nM, respectively. The resistance index of 1-12228 crizotinib-resistant cells was 10.20. Crizotinib induced apoptosis in H2228 cells and reduced the levels of p-ALK, p-STAT3, and survivin. In contrast, no changes in the levels of p-ALK, p-STAT3, and survivin were observed in H2228 crizotinib-resistant cells. The mutations 2067G--,A and 2182G--,C in EML4-ALK were present in the H2228 crizotinib-resistant cells. Conclusion Crizotinib decreased the viability of H2228 cells in a dose- and time-dependent manner. In the STAT3/survivin pathway, downregulation of p-ALK, p-STAT3, and survivin might contribute to crizo- tinib-induced apoptosis in H2228 ceils. However, the STAT3/survivin pathway in H2228 crizotinib-resistant cells was unaffected by crizotinib treatment. Acquired resistance in H2228 cells might be related to ALK mutations.
