Background:Colorectal cancer(CRC)represents a substantial risk to public health.Bevacizumab,thefirst US FDA-approved antiangiogenic drug(AAD)for human CRC treatment,faces resistance in patients.The role of lipid metabo...Background:Colorectal cancer(CRC)represents a substantial risk to public health.Bevacizumab,thefirst US FDA-approved antiangiogenic drug(AAD)for human CRC treatment,faces resistance in patients.The role of lipid metabolism,particularly through OPA3-regulated lipid droplet production,in overcoming this resistance is under investigation.Methods:The protein expression pattern of OPA3 in CRC primary/normal tissues was evaluated by bioinformatics analysis.OPA3-overexpressed SW-480 and HCT-116 cell lines were established,and bevacizumab resistance and OPA3 effects on cell malignancy were examined.OPA3 protein/mRNA expression and lipid droplet-related genes were measured with Western blot and qRT-PCR.OPA3 subcellular localization was detected using immunofluorescence.Proliferation and apoptosis were assessed via colony formation andflow cytometry.Tube formation assays were conducted to assess the angiogenic potential of human umbilical vein endothelial cells(HUVECs).Lipid analysis was used to measure the phosphatidylcholine(PC)and lysophosphatidylcholine(LPC)levels in CRC cells.Results:Bioinformatics analysis revealed that OPA3 was downregulated in CRC.Overexpression of OPA3 inhibited CRC cell proliferation,stimulated apoptosis,and suppressed the angiogenic ability of HUVECs.OPA3 effectively reversed the resistance of CRC cells to bevacizumab and decreased lipid droplet production in CRC cells.Additionally,OPA3 reversed the bevacizumab-induced lipid droplet production in CRC cells,thereby increasing CRC cell sensitivity to bevacizumab treatment.Conclusion:This study suggests that OPA3 modulates lipid metabolism in CRC cells and reduces resistance to bevacizumab in CRC cells.Therefore,OPA3 may be a potential therapeutic target against the AAD resistance in CRC.展开更多
3型视神经萎缩蛋白(Optic atrophy type 3,OPA3)普遍存在于真菌、植物和动物中,但它在植物中的功能尚不清楚。本研究构建了番茄OPA3-like(SlOPA3L)基因的过表达载体35S∶∶SlOPA3L并遗传转化烟草。与野生型烟草叶片相比,过量表达SlOPA3...3型视神经萎缩蛋白(Optic atrophy type 3,OPA3)普遍存在于真菌、植物和动物中,但它在植物中的功能尚不清楚。本研究构建了番茄OPA3-like(SlOPA3L)基因的过表达载体35S∶∶SlOPA3L并遗传转化烟草。与野生型烟草叶片相比,过量表达SlOPA3L基因的转基因烟草的子叶呈花叶状、部分组织失绿,转基因烟草的少数早期真叶呈花叶表型,随着叶片发育后又逐渐恢复正常。转基因烟草的叶片细胞表型分析表明:真叶花叶的叶片的海绵组织和栅栏组织发育受到显著的影响。这些结果表明:SlOPA3L基因可能影植物叶片细胞的分裂和参与植物叶片发育早期的叶绿体形成。展开更多
Introduction. In 1961, Garcin et al. described a family with several members affected with optic atrophy associated with cataract, and neurological symptoms. The authors believed this condition to be distinct from oth...Introduction. In 1961, Garcin et al. described a family with several members affected with optic atrophy associated with cataract, and neurological symptoms. The authors believed this condition to be distinct from other diseases known at that time, e.g. the Behr syndrome, Marinesco-Sj gren syndrome and Friedreich’s ataxia. Method. This family was followed over a period of 40 years and genes known to be responsible for optic atrophy were sequenced. Results. The G277A mutation of OPA3 gene was responsible for this familial disease. Discussion. A new clinical entity is identified: autosomal dominant optic atrophy and cataract, due to a heterozygous mutation of the OPA3 gene, a nuclear gene encoding a mitochondrial protein.展开更多
目的筛选肿瘤细胞中与生物钟分子振荡系统的重要组成部分PERl蛋白（period circadian protein homolog1）相互作用的蛋白质分子，为生物钟基因在肿瘤发生发展过程中的功能研究提供条件。方法应用细菌双杂交技术与人宫颈癌Hela细胞cDNA...目的筛选肿瘤细胞中与生物钟分子振荡系统的重要组成部分PERl蛋白（period circadian protein homolog1）相互作用的蛋白质分子，为生物钟基因在肿瘤发生发展过程中的功能研究提供条件。方法应用细菌双杂交技术与人宫颈癌Hela细胞cDNA文库，以pBT为载体，构建pBT-PERl融合表达诱饵质粒，与pTRG连接的人宫颈癌HeIa细胞cDNA文库质粒共转化双杂交系统报告菌株，利用培养基的特殊选择性，筛选出阳性克隆并测序分析。结果筛选出14个蛋白编码基因，其中4个包含完整的蛋白编码序列，包括与线粒体动力学相关的OPA3蛋白，与铜代谢相关的CUTA蛋白，与细胞运动、定位等细胞事件相关的蛋白，与物质合成、代谢等生物化学反应相关的蛋白，还有在信号转导中发挥作用的相关蛋白。结论肿瘤细胞中与PER1蛋白存在潜在相互作用新蛋白的筛选和鉴定为研究人生物钟基因PERI在肿瘤发生发展中的功能提供了条件。展开更多
文摘Background:Colorectal cancer(CRC)represents a substantial risk to public health.Bevacizumab,thefirst US FDA-approved antiangiogenic drug(AAD)for human CRC treatment,faces resistance in patients.The role of lipid metabolism,particularly through OPA3-regulated lipid droplet production,in overcoming this resistance is under investigation.Methods:The protein expression pattern of OPA3 in CRC primary/normal tissues was evaluated by bioinformatics analysis.OPA3-overexpressed SW-480 and HCT-116 cell lines were established,and bevacizumab resistance and OPA3 effects on cell malignancy were examined.OPA3 protein/mRNA expression and lipid droplet-related genes were measured with Western blot and qRT-PCR.OPA3 subcellular localization was detected using immunofluorescence.Proliferation and apoptosis were assessed via colony formation andflow cytometry.Tube formation assays were conducted to assess the angiogenic potential of human umbilical vein endothelial cells(HUVECs).Lipid analysis was used to measure the phosphatidylcholine(PC)and lysophosphatidylcholine(LPC)levels in CRC cells.Results:Bioinformatics analysis revealed that OPA3 was downregulated in CRC.Overexpression of OPA3 inhibited CRC cell proliferation,stimulated apoptosis,and suppressed the angiogenic ability of HUVECs.OPA3 effectively reversed the resistance of CRC cells to bevacizumab and decreased lipid droplet production in CRC cells.Additionally,OPA3 reversed the bevacizumab-induced lipid droplet production in CRC cells,thereby increasing CRC cell sensitivity to bevacizumab treatment.Conclusion:This study suggests that OPA3 modulates lipid metabolism in CRC cells and reduces resistance to bevacizumab in CRC cells.Therefore,OPA3 may be a potential therapeutic target against the AAD resistance in CRC.
文摘3型视神经萎缩蛋白(Optic atrophy type 3,OPA3)普遍存在于真菌、植物和动物中,但它在植物中的功能尚不清楚。本研究构建了番茄OPA3-like(SlOPA3L)基因的过表达载体35S∶∶SlOPA3L并遗传转化烟草。与野生型烟草叶片相比,过量表达SlOPA3L基因的转基因烟草的子叶呈花叶状、部分组织失绿,转基因烟草的少数早期真叶呈花叶表型,随着叶片发育后又逐渐恢复正常。转基因烟草的叶片细胞表型分析表明:真叶花叶的叶片的海绵组织和栅栏组织发育受到显著的影响。这些结果表明:SlOPA3L基因可能影植物叶片细胞的分裂和参与植物叶片发育早期的叶绿体形成。
文摘Introduction. In 1961, Garcin et al. described a family with several members affected with optic atrophy associated with cataract, and neurological symptoms. The authors believed this condition to be distinct from other diseases known at that time, e.g. the Behr syndrome, Marinesco-Sj gren syndrome and Friedreich’s ataxia. Method. This family was followed over a period of 40 years and genes known to be responsible for optic atrophy were sequenced. Results. The G277A mutation of OPA3 gene was responsible for this familial disease. Discussion. A new clinical entity is identified: autosomal dominant optic atrophy and cataract, due to a heterozygous mutation of the OPA3 gene, a nuclear gene encoding a mitochondrial protein.
文摘目的筛选肿瘤细胞中与生物钟分子振荡系统的重要组成部分PERl蛋白（period circadian protein homolog1）相互作用的蛋白质分子，为生物钟基因在肿瘤发生发展过程中的功能研究提供条件。方法应用细菌双杂交技术与人宫颈癌Hela细胞cDNA文库，以pBT为载体，构建pBT-PERl融合表达诱饵质粒，与pTRG连接的人宫颈癌HeIa细胞cDNA文库质粒共转化双杂交系统报告菌株，利用培养基的特殊选择性，筛选出阳性克隆并测序分析。结果筛选出14个蛋白编码基因，其中4个包含完整的蛋白编码序列，包括与线粒体动力学相关的OPA3蛋白，与铜代谢相关的CUTA蛋白，与细胞运动、定位等细胞事件相关的蛋白，与物质合成、代谢等生物化学反应相关的蛋白，还有在信号转导中发挥作用的相关蛋白。结论肿瘤细胞中与PER1蛋白存在潜在相互作用新蛋白的筛选和鉴定为研究人生物钟基因PERI在肿瘤发生发展中的功能提供了条件。
