AIM: To investigate the association between the configurational and compositional changes of nuclear matrix and the differentiation of carcinoma cells. METHODS: Cells cultured with or without 5 × 10^-3 mmol/L o...AIM: To investigate the association between the configurational and compositional changes of nuclear matrix and the differentiation of carcinoma cells. METHODS: Cells cultured with or without 5 × 10^-3 mmol/L of hexamethylene bisacetamide (HMBA) on Nickel grids were treated by selective extraction and prepared for whole mount observation under electron microscopy. The samples were examined under transmission electron microscope. Nuclear matrix proteins were selectively extracted and subjected to subcellular proteomics study. The protein expression patterns were analyzed by PDQuest software. Spots of differentially expressed nuclear matrix proteins were excised and subjected to in situ digestion with trypsin. The peptides were analyzed by matrix-assisted laser- desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Data were submitted for database searching using Mascot tool (www.matrixscience.com). RESULTS: The nuclear matrix (NM) and intermediate filament (IF) in SMMC-7721 hepatocarcinoma cells were found relatively sparse and arranged irregularly. The nuclear lamina was non-uniform, and two kinds of filaments were not tightly connected. After induction for differentiation by HMBA, the NM-IF filaments were concentrated and distributed uniformly. The heterogeneous population of filaments, including highly branched utrathin filaments could also be seen in the regular meshwork. The connection between the two kinds of filaments and the relatively thin, condensed and sharply demarcated lamina composed of intermediate- sized filaments was relatively fastened. Meanwhile, 21 NM proteins changed remarkably during SMMC-7721 cell differentiation. Four proteins, i.e. mutant Pystl, hypothetical protein, nucleophosminl, and LBP were downregulated, whereas four other proteins, eIF6, p44 subunit, 13-tubulin, and SIN3B were upregulated with the last one, SR2/ASF found only in the differentiated SMMC-7721 cells. CONCLUSION: The induced differentiation of SMMC-7721 cells by HMBA is accompanied by the configurational changes of nuclear matrix-intermediate filament (NM-IF) system and the compositional changes of nuclear matrix protein expression. These changes may be important morphological or functional indications of the cancer cell reversion.展开更多
Traditional Chinese medicine(TCM)was formed two thousand years ago,and developed in the following centuries.TCM has a unique way of looking at health and illness.Zheng(syndrome)is the basic unit and key term in TCM th...Traditional Chinese medicine(TCM)was formed two thousand years ago,and developed in the following centuries.TCM has a unique way of looking at health and illness.Zheng(syndrome)is the basic unit and key term in TCM theory.Zheng is the pathological generalization of a certain stage in the development of disease.Treatment based on syndrome differentiation and disease differentiation of TCM is the basic principle to know and treat diseases,and a kind of special method to study diseases in TCM.In Han Dynasty(200 AD),doctor Zhang Zhongjing proposed Liujing Bianzheng(the differential diagnosis in accordance with the theory of six channels)to diagnose and treat diseases in Treatise on Febrile and Miscellaneous Diseases.In other words,he divided diseases into six channels diseases based on patient's Zhengs.Six channels diseases almost include all of the disease according to his classification standards.In addition,there are development rules in the progress of six channels diseases.Therefore,we hypothesized that combination of six channels disease and Zheng on the basic of differentiation of six channels is a new pattern of diagnosis and treatment diseases in TCM,which is beneficial to the understanding of a certain disease and makes disease treatment more convenient,fast and effective.展开更多
基金Supported by the National Natural Science Foundation of China, No. 30470877
文摘AIM: To investigate the association between the configurational and compositional changes of nuclear matrix and the differentiation of carcinoma cells. METHODS: Cells cultured with or without 5 × 10^-3 mmol/L of hexamethylene bisacetamide (HMBA) on Nickel grids were treated by selective extraction and prepared for whole mount observation under electron microscopy. The samples were examined under transmission electron microscope. Nuclear matrix proteins were selectively extracted and subjected to subcellular proteomics study. The protein expression patterns were analyzed by PDQuest software. Spots of differentially expressed nuclear matrix proteins were excised and subjected to in situ digestion with trypsin. The peptides were analyzed by matrix-assisted laser- desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Data were submitted for database searching using Mascot tool (www.matrixscience.com). RESULTS: The nuclear matrix (NM) and intermediate filament (IF) in SMMC-7721 hepatocarcinoma cells were found relatively sparse and arranged irregularly. The nuclear lamina was non-uniform, and two kinds of filaments were not tightly connected. After induction for differentiation by HMBA, the NM-IF filaments were concentrated and distributed uniformly. The heterogeneous population of filaments, including highly branched utrathin filaments could also be seen in the regular meshwork. The connection between the two kinds of filaments and the relatively thin, condensed and sharply demarcated lamina composed of intermediate- sized filaments was relatively fastened. Meanwhile, 21 NM proteins changed remarkably during SMMC-7721 cell differentiation. Four proteins, i.e. mutant Pystl, hypothetical protein, nucleophosminl, and LBP were downregulated, whereas four other proteins, eIF6, p44 subunit, 13-tubulin, and SIN3B were upregulated with the last one, SR2/ASF found only in the differentiated SMMC-7721 cells. CONCLUSION: The induced differentiation of SMMC-7721 cells by HMBA is accompanied by the configurational changes of nuclear matrix-intermediate filament (NM-IF) system and the compositional changes of nuclear matrix protein expression. These changes may be important morphological or functional indications of the cancer cell reversion.
文摘Traditional Chinese medicine(TCM)was formed two thousand years ago,and developed in the following centuries.TCM has a unique way of looking at health and illness.Zheng(syndrome)is the basic unit and key term in TCM theory.Zheng is the pathological generalization of a certain stage in the development of disease.Treatment based on syndrome differentiation and disease differentiation of TCM is the basic principle to know and treat diseases,and a kind of special method to study diseases in TCM.In Han Dynasty(200 AD),doctor Zhang Zhongjing proposed Liujing Bianzheng(the differential diagnosis in accordance with the theory of six channels)to diagnose and treat diseases in Treatise on Febrile and Miscellaneous Diseases.In other words,he divided diseases into six channels diseases based on patient's Zhengs.Six channels diseases almost include all of the disease according to his classification standards.In addition,there are development rules in the progress of six channels diseases.Therefore,we hypothesized that combination of six channels disease and Zheng on the basic of differentiation of six channels is a new pattern of diagnosis and treatment diseases in TCM,which is beneficial to the understanding of a certain disease and makes disease treatment more convenient,fast and effective.