沉默信息调节因子2(silent information regulator 2,SIRT2)作为sirtuins蛋白家族成员广泛存在于真核生物之中,是具备调节生物生成和代谢调控过程作用的Ⅲ类去乙酰化酶,参与调控细胞分化、新陈代谢、衰老、DNA损伤修复、维持基因组完整...沉默信息调节因子2(silent information regulator 2,SIRT2)作为sirtuins蛋白家族成员广泛存在于真核生物之中,是具备调节生物生成和代谢调控过程作用的Ⅲ类去乙酰化酶,参与调控细胞分化、新陈代谢、衰老、DNA损伤修复、维持基因组完整性以及肿瘤的发生等。SIRT2的异常表达与多种肿瘤的发生发展具有密切关系,例如其在乳腺癌、宫颈癌、肺癌等恶性肿瘤方面的研究进展,SIRT2在发挥原癌基因作用的基础上,也发挥抑癌基因的作用。该文主要就SIRT2在肿瘤发生发展中的分子机制及其研究进展进行综述,以期为研究者开发肿瘤治疗药物提供新视野。展开更多
Objective: To demonstrate whether the expression of silent mating type information regulation 2 homolog 1 (SIRT1) affects the level of TGF-β1 and Smad3 in HEK293 cells through regulating mTOR. Methods: First, recombi...Objective: To demonstrate whether the expression of silent mating type information regulation 2 homolog 1 (SIRT1) affects the level of TGF-β1 and Smad3 in HEK293 cells through regulating mTOR. Methods: First, recombinant plasmids DNA (rSIRT1) and siRNA targeting SIRT1 were constructed which were transfected into Human Embryonic Kidney 293 cell (HEK293) cells, respectively. Then, the generation of intracellular ROS in cells was examined by flow cytometry using the oxidation-sensitive probe. Last, the expressions of TGF-β1, smad3, P53, mTOR, p-mTOR, LC3-I and LC3-II in cells were detected to observe the effect of SIRT1 on TGF-β1 Pathway by western blot analysis. Results: We demonstrated that overexpressing of SIRT1 may decrease TGF-β1 and Smad3 expression in HEK293 cells through regulating mTOR. In addition, the result is the opposite when SIRT1 was silent in HEK293 cells. Conclusions: SIRT1 is closely related to TGF-β1/Smad3 pathway that correlates with the regulation of mTOR and ROS generation and causes diabetic nephropathy. The available evidence implies that SIRT1 has great potential as a clinical target for the prevention and treatment of renal fibrosis in the development of DN.展开更多
Objective:To study the effects of silent information regulator of transcription 2 (SIRT2) on inflammatory response and bone destruction in cartilage tissue of osteoarthritis.Methods: A total of 200 patients who underw...Objective:To study the effects of silent information regulator of transcription 2 (SIRT2) on inflammatory response and bone destruction in cartilage tissue of osteoarthritis.Methods: A total of 200 patients who underwent knee replacement due to knee osteoarthritis in Kashgar Prefecture First People's Hospital between September 2014 and September 2017 were selected as the osteoarthritis (OA) group of the research, and 80 patients who underwent knee replacement or meniscus operation due to trauma in Kashgar Prefecture First People's Hospital during the same period were selected as the control group. Articular cartilage tissue was collected after surgery to measure the expression of SIRT2 and bone destruction-related apoptosis molecules as well as the levels of inflammatory response molecules and bone destruction-related collagen metabolism molecules.Results: SIRT2 and Bcl-2 mRNA expression as well as SOX9 and Col-II levels in articular cartilage tissue of OA group were significantly lower than those of control group whereas TNF-α, bFGF, NO, IP-10, CCL2, PAR-2,β-catenin, OPN and MMP13 levels as well as Fas, GRP78, ATF6 and Caspase-3 mRNA expression were significantly higher than those of control group;SIRT2 mRNA expression in articular cartilage tissue of OA group was positively correlated with Bcl-2 mRNA expression as well as SOX9 and Col-II levels, and negatively correlated with TNF-α, bFGF, NO, IP-10, CCL2, PAR-2,β-catenin, OPN and MMP13 levels as well as Fas, GRP78, ATF6 and Caspase-3 mRNA expression.Conclusion: The lowly expressed SIRT2 in cartilage of osteoarthritis can aggravate inflammatory response and bone destruction.展开更多
AIM： To detect the expression of miR-211 in age-related cataract tissue, explore the effects of miR-211 on lens epithelial cell proliferation and apoptosis, and identify its target gene.METHODS： This study used real...AIM： To detect the expression of miR-211 in age-related cataract tissue, explore the effects of miR-211 on lens epithelial cell proliferation and apoptosis, and identify its target gene.METHODS： This study used real-time quantitative polymerase chain reaction（RT-q PCR） to measure the expression of miR-211 and its predicted target gene [silent matingtype information regulation 2 homolog 1（SIRT1）] in 46 anterior lens capsules collected from age-related cataract patients. Human lens epithelial cell line（SRA01/04） cells were transfected with either miR-211 mimics, mimic controls, miR-211 inhibitors or inhibitor controls, 72 h after transfection, miR NA and protein expression of SIRT1 were measured using RT-qP CR and Western blotting; then cells were exposed to 200 μmol/L H2O2 for 1h, whereupon cell viability was measured by MTS assay, caspase-3 assay was performed. Dual luciferase reporter assay was performed to verify the relationship between miR-211 of SIRT1.RESULTS： Compared to the control group, expression of miR-211 was significantly increased（P〈0.001）, the miR NA and protein expression of SIRT1 were significantly decreased（P〈0.001） in the anterior lens capsules of patients with age-related cataracts. Relative to the control group, SIRT1 miR NA and protein levels in the miR-211 mimic group were significantly reduced, cell proliferation activity significantly decreased, and caspase-3 activity was significantly increased（P〈0.001）. In the miR-211 inhibitor group, SIRT1 miRNA and protein expression were significantly increased, cell proliferation activity significantly increased, and caspase-3 activity was significantly decreased（P〈0.001）. A dual luciferase reporter assay confirmed that SIRT1 is a direct target of miR-211.CONCLUSION： miR-211 is highly expressed in the anterior lens capsules of patients with age-related cataracts. By negatively regulating the expression of SIRT1, miR-211 promotes lens epithelial cell apoptosis and inhibits lens epithelial cell proliferation.展开更多
目的:通过生物信息学方法预测和分析人沉默信息调节因子2(Silent Information Regulator 2,SIRT2)基因结构和功能,利用分子生物学方法构建SIRT2重组质粒并进行原核表达。方法:利用NCBI、STRING、ProtParam、Genecards和SWISS-MODEL等软...目的:通过生物信息学方法预测和分析人沉默信息调节因子2(Silent Information Regulator 2,SIRT2)基因结构和功能,利用分子生物学方法构建SIRT2重组质粒并进行原核表达。方法:利用NCBI、STRING、ProtParam、Genecards和SWISS-MODEL等软件和数据库对SIRT2蛋白的理化性质、跨膜结构域、亚细胞定位、翻译后修饰、二级结构、三级结构及相互作用网络等生物学特性进行了预测分析,采用PCR扩增技术获得SIRT2基因的cDNA序列全长,通过同源重组的方法构建p ET-28aSIRT2重组表达质粒并进行原核表达。结果:人SIRT2蛋白由389个氨基酸组成,是一种呈酸性、不稳定的、亲水的胞内蛋白质。SIRT2主要分布在细胞质,具有38个潜在的磷酸化修饰位点,其二级结构主要是无规则卷曲。成功构建了重组质粒p ET-28aSIRT2,并通过IPTG诱导大肠杆菌BL21后成功获得SIRT2蛋白的表达,SIRT2主要存在于细菌裂解液离心之后的沉淀中。结论:本研究成功克隆了人SIRT2的编码区,结合多种生物信息软件对SIRT2的性质和功能进行了了解,为进一步研究SIRT2蛋白的生物学功能和肿瘤的靶向治疗作用机制提供思路。展开更多
文摘沉默信息调节因子2(silent information regulator 2,SIRT2)作为sirtuins蛋白家族成员广泛存在于真核生物之中,是具备调节生物生成和代谢调控过程作用的Ⅲ类去乙酰化酶,参与调控细胞分化、新陈代谢、衰老、DNA损伤修复、维持基因组完整性以及肿瘤的发生等。SIRT2的异常表达与多种肿瘤的发生发展具有密切关系,例如其在乳腺癌、宫颈癌、肺癌等恶性肿瘤方面的研究进展,SIRT2在发挥原癌基因作用的基础上,也发挥抑癌基因的作用。该文主要就SIRT2在肿瘤发生发展中的分子机制及其研究进展进行综述,以期为研究者开发肿瘤治疗药物提供新视野。
文摘Objective: To demonstrate whether the expression of silent mating type information regulation 2 homolog 1 (SIRT1) affects the level of TGF-β1 and Smad3 in HEK293 cells through regulating mTOR. Methods: First, recombinant plasmids DNA (rSIRT1) and siRNA targeting SIRT1 were constructed which were transfected into Human Embryonic Kidney 293 cell (HEK293) cells, respectively. Then, the generation of intracellular ROS in cells was examined by flow cytometry using the oxidation-sensitive probe. Last, the expressions of TGF-β1, smad3, P53, mTOR, p-mTOR, LC3-I and LC3-II in cells were detected to observe the effect of SIRT1 on TGF-β1 Pathway by western blot analysis. Results: We demonstrated that overexpressing of SIRT1 may decrease TGF-β1 and Smad3 expression in HEK293 cells through regulating mTOR. In addition, the result is the opposite when SIRT1 was silent in HEK293 cells. Conclusions: SIRT1 is closely related to TGF-β1/Smad3 pathway that correlates with the regulation of mTOR and ROS generation and causes diabetic nephropathy. The available evidence implies that SIRT1 has great potential as a clinical target for the prevention and treatment of renal fibrosis in the development of DN.
文摘Objective:To study the effects of silent information regulator of transcription 2 (SIRT2) on inflammatory response and bone destruction in cartilage tissue of osteoarthritis.Methods: A total of 200 patients who underwent knee replacement due to knee osteoarthritis in Kashgar Prefecture First People's Hospital between September 2014 and September 2017 were selected as the osteoarthritis (OA) group of the research, and 80 patients who underwent knee replacement or meniscus operation due to trauma in Kashgar Prefecture First People's Hospital during the same period were selected as the control group. Articular cartilage tissue was collected after surgery to measure the expression of SIRT2 and bone destruction-related apoptosis molecules as well as the levels of inflammatory response molecules and bone destruction-related collagen metabolism molecules.Results: SIRT2 and Bcl-2 mRNA expression as well as SOX9 and Col-II levels in articular cartilage tissue of OA group were significantly lower than those of control group whereas TNF-α, bFGF, NO, IP-10, CCL2, PAR-2,β-catenin, OPN and MMP13 levels as well as Fas, GRP78, ATF6 and Caspase-3 mRNA expression were significantly higher than those of control group;SIRT2 mRNA expression in articular cartilage tissue of OA group was positively correlated with Bcl-2 mRNA expression as well as SOX9 and Col-II levels, and negatively correlated with TNF-α, bFGF, NO, IP-10, CCL2, PAR-2,β-catenin, OPN and MMP13 levels as well as Fas, GRP78, ATF6 and Caspase-3 mRNA expression.Conclusion: The lowly expressed SIRT2 in cartilage of osteoarthritis can aggravate inflammatory response and bone destruction.
基金Supported by the National Natural Science Foundation of China(No.81170836No.81570838)+1 种基金the Natural Science Foundation of Liaoning Province,China(No.2015020474)the Liaoning Provincial Hospital Program for Building Treatment Capacity in Key Clinical Departments(No.LNCCC-D15-2015)
文摘AIM： To detect the expression of miR-211 in age-related cataract tissue, explore the effects of miR-211 on lens epithelial cell proliferation and apoptosis, and identify its target gene.METHODS： This study used real-time quantitative polymerase chain reaction（RT-q PCR） to measure the expression of miR-211 and its predicted target gene [silent matingtype information regulation 2 homolog 1（SIRT1）] in 46 anterior lens capsules collected from age-related cataract patients. Human lens epithelial cell line（SRA01/04） cells were transfected with either miR-211 mimics, mimic controls, miR-211 inhibitors or inhibitor controls, 72 h after transfection, miR NA and protein expression of SIRT1 were measured using RT-qP CR and Western blotting; then cells were exposed to 200 μmol/L H2O2 for 1h, whereupon cell viability was measured by MTS assay, caspase-3 assay was performed. Dual luciferase reporter assay was performed to verify the relationship between miR-211 of SIRT1.RESULTS： Compared to the control group, expression of miR-211 was significantly increased（P〈0.001）, the miR NA and protein expression of SIRT1 were significantly decreased（P〈0.001） in the anterior lens capsules of patients with age-related cataracts. Relative to the control group, SIRT1 miR NA and protein levels in the miR-211 mimic group were significantly reduced, cell proliferation activity significantly decreased, and caspase-3 activity was significantly increased（P〈0.001）. In the miR-211 inhibitor group, SIRT1 miRNA and protein expression were significantly increased, cell proliferation activity significantly increased, and caspase-3 activity was significantly decreased（P〈0.001）. A dual luciferase reporter assay confirmed that SIRT1 is a direct target of miR-211.CONCLUSION： miR-211 is highly expressed in the anterior lens capsules of patients with age-related cataracts. By negatively regulating the expression of SIRT1, miR-211 promotes lens epithelial cell apoptosis and inhibits lens epithelial cell proliferation.
