目的探究溶质载体家族7成员5(solute carrier family 7 member 5,SLC7A5)促进转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导的人肺成纤维细胞活化增殖的作用。方法人肺成纤维细胞用TGF-β1孵育24h,使用PCR-Array方法...目的探究溶质载体家族7成员5(solute carrier family 7 member 5,SLC7A5)促进转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导的人肺成纤维细胞活化增殖的作用。方法人肺成纤维细胞用TGF-β1孵育24h,使用PCR-Array方法筛选靶基因。使用沉默慢病毒sh-SLC和重表达慢病毒r-SLC转染成纤维细胞,利用Western Blot实验和qRT-PCR实验观察目标基因SLC7A5和成纤维细胞活化指标α平滑肌肌动蛋白(alpha smooth muscle actin,α-SMA)、成纤维细胞特异性蛋白(fibroblast specific protein,FSP)、Ⅰ型胶原(typeⅠcollagen,COL-1)合成。qRT-PCR实验检测mTOR信号通路分子。CCK-8方法检测各组成纤维细胞增殖速度。流式细胞术检测各组细胞周期变化情况。结果与对照组PBS的细胞相比,TGF-β1实验组的HLFs中SLC7A5高表达,沉默SLC7A5基因降低SLC7A5、α-SMA、FSP和COL-1表达量以及mTOR和4EBP1的转录水平,抑制成纤维细胞增殖速度,G1期比例增加,S期降低,而回复实验表明重新表达SLC7A5基因的r-SLC能够逆转以上现象。结论SLC7A5可能通过mTOR信号通路促进TGF-β1诱导的人肺成纤维细胞活化增殖,SLC7A5有望成为肺纤维化疾病中抑制成纤维细胞活化增殖的靶点。展开更多
Progressive photoreceptor cell death is one of the main pathological features of age-related macular degeneration and eventually leads to vision loss.Ferroptosis has been demonstrated to be associated with retinal deg...Progressive photoreceptor cell death is one of the main pathological features of age-related macular degeneration and eventually leads to vision loss.Ferroptosis has been demonstrated to be associated with retinal degenerative diseases.However,the molecular mechanisms underlying ferroptosis and photoreceptor cell death in age-related macular degeneration remain largely unexplored.Bioinformatics and biochemical analyses in this study revealed xC^(–),solute carrier family 7 member 11-regulated ferroptosis as the predominant pathological process of photoreceptor cell degeneration in a light-induced dry age-related macular degeneration mouse model.This process involves the nuclear factor-erythroid factor 2-related factor 2-solute carrier family 7 member 11-glutathione peroxidase 4 signaling pathway,through which cystine depletion,iron ion accumulation,and enhanced lipid peroxidation ultimately lead to photoreceptor cell death and subsequent visual function impairment.We demonstrated that solute carrier family 7 member 11 overexpression blocked this process by inhibiting oxidative stress in vitro and in vivo.Conversely,solute carrier family 7 member 11 knockdown or the solute carrier family 7 member 11 inhibitor sulfasalazine and ferroptosis-inducing agent erastin aggravated H_(2)O_(2)-induced ferroptosis of 661W cells.These findings indicate solute carrier family 7 member 11 may be a potential therapeutic target for patients with retinal degenerative diseases including age-related macular degeneration.展开更多
Objective:Breast cancer is the most common malignancy in women and is characterized by a high recurrence rate that severely impacts patient survival.Regulatory T cells(Tregs)in the tumor microenvironment(TME)promote i...Objective:Breast cancer is the most common malignancy in women and is characterized by a high recurrence rate that severely impacts patient survival.Regulatory T cells(Tregs)in the tumor microenvironment(TME)promote immune evasion and metastasis,increasing recurrence risk.This study determined how the epigenetic regulators,DNMT3A and METTL7A,modulate Treg infiltration via the DDR1/STAT3/CXCL5 axis and influence breast cancer recurrence and prognosis.Methods:RNA sequencing(RNA-seq)was used to identify differentially expressed genes(DEGs),followed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment.Machine learning algorithms,including least absolute shrinkage and selection operator(LASSO),supported vector machine-recursive feature elimination(SVM-RFE)and ElasticNet identified DDR1 as a key gene.Validation included RT-qPCR,western blot,MSP,MeRIP-qPCR,and Co-IP to assess epigenetic regulation.Functional assays(CCK-8,Transwell,and Treg differentiation/chemotaxis)and xenograft models evaluated the role of DDR1 in tumor progression and recurrence.Results:DNMT3A upregulated DDR1 via DNA methylation,while METTL7A enhanced DDR1 mRNA stability via m6A modification.Co-regulation activated the DDR1/STAT3/CXCL5 axis,which boosted cancer cell proliferation,migration,and invasion.CXCL5 secretion increased Treg infiltration and accelerated tumor growth in vivo.DDR1 silencing reversed these effects,confirming that DDR1 has a pivotal role in breast cancer recurrence.Conclusion:DNMT3A and METTL7A were shown to cooperatively regulate DDR1 via DNA/m6A methylation,which drives Tregmediated immune suppression and recurrence.This study provided novel insights and therapeutic targets for breast cancer prognosis and treatment.展开更多
文摘目的探究溶质载体家族7成员5(solute carrier family 7 member 5,SLC7A5)促进转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导的人肺成纤维细胞活化增殖的作用。方法人肺成纤维细胞用TGF-β1孵育24h,使用PCR-Array方法筛选靶基因。使用沉默慢病毒sh-SLC和重表达慢病毒r-SLC转染成纤维细胞,利用Western Blot实验和qRT-PCR实验观察目标基因SLC7A5和成纤维细胞活化指标α平滑肌肌动蛋白(alpha smooth muscle actin,α-SMA)、成纤维细胞特异性蛋白(fibroblast specific protein,FSP)、Ⅰ型胶原(typeⅠcollagen,COL-1)合成。qRT-PCR实验检测mTOR信号通路分子。CCK-8方法检测各组成纤维细胞增殖速度。流式细胞术检测各组细胞周期变化情况。结果与对照组PBS的细胞相比,TGF-β1实验组的HLFs中SLC7A5高表达,沉默SLC7A5基因降低SLC7A5、α-SMA、FSP和COL-1表达量以及mTOR和4EBP1的转录水平,抑制成纤维细胞增殖速度,G1期比例增加,S期降低,而回复实验表明重新表达SLC7A5基因的r-SLC能够逆转以上现象。结论SLC7A5可能通过mTOR信号通路促进TGF-β1诱导的人肺成纤维细胞活化增殖,SLC7A5有望成为肺纤维化疾病中抑制成纤维细胞活化增殖的靶点。
基金supported by the National Natural Science Foundation of China,Nos.82171076(to XS)and U22A20311(to XS),82101168(to TL)Shanghai Science and technology Innovation Action Plan,No.23Y11901300(to JS)+1 种基金Science and Technology Commission of Shanghai Municipality,No.21ZR1451500(to TL)Shanghai Pujiang Program,No.22PJ1412200(to BY)。
文摘Progressive photoreceptor cell death is one of the main pathological features of age-related macular degeneration and eventually leads to vision loss.Ferroptosis has been demonstrated to be associated with retinal degenerative diseases.However,the molecular mechanisms underlying ferroptosis and photoreceptor cell death in age-related macular degeneration remain largely unexplored.Bioinformatics and biochemical analyses in this study revealed xC^(–),solute carrier family 7 member 11-regulated ferroptosis as the predominant pathological process of photoreceptor cell degeneration in a light-induced dry age-related macular degeneration mouse model.This process involves the nuclear factor-erythroid factor 2-related factor 2-solute carrier family 7 member 11-glutathione peroxidase 4 signaling pathway,through which cystine depletion,iron ion accumulation,and enhanced lipid peroxidation ultimately lead to photoreceptor cell death and subsequent visual function impairment.We demonstrated that solute carrier family 7 member 11 overexpression blocked this process by inhibiting oxidative stress in vitro and in vivo.Conversely,solute carrier family 7 member 11 knockdown or the solute carrier family 7 member 11 inhibitor sulfasalazine and ferroptosis-inducing agent erastin aggravated H_(2)O_(2)-induced ferroptosis of 661W cells.These findings indicate solute carrier family 7 member 11 may be a potential therapeutic target for patients with retinal degenerative diseases including age-related macular degeneration.
基金supported by the National Natural Science Foundation of China(Grant No.82060479)Key Research and Development Program of Ningxia Hui Autonomous Region(Grant No.2021BEG03062)Ningxia Natural Science Fund Key Project(Grant No.2024AAC02080).
文摘Objective:Breast cancer is the most common malignancy in women and is characterized by a high recurrence rate that severely impacts patient survival.Regulatory T cells(Tregs)in the tumor microenvironment(TME)promote immune evasion and metastasis,increasing recurrence risk.This study determined how the epigenetic regulators,DNMT3A and METTL7A,modulate Treg infiltration via the DDR1/STAT3/CXCL5 axis and influence breast cancer recurrence and prognosis.Methods:RNA sequencing(RNA-seq)was used to identify differentially expressed genes(DEGs),followed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment.Machine learning algorithms,including least absolute shrinkage and selection operator(LASSO),supported vector machine-recursive feature elimination(SVM-RFE)and ElasticNet identified DDR1 as a key gene.Validation included RT-qPCR,western blot,MSP,MeRIP-qPCR,and Co-IP to assess epigenetic regulation.Functional assays(CCK-8,Transwell,and Treg differentiation/chemotaxis)and xenograft models evaluated the role of DDR1 in tumor progression and recurrence.Results:DNMT3A upregulated DDR1 via DNA methylation,while METTL7A enhanced DDR1 mRNA stability via m6A modification.Co-regulation activated the DDR1/STAT3/CXCL5 axis,which boosted cancer cell proliferation,migration,and invasion.CXCL5 secretion increased Treg infiltration and accelerated tumor growth in vivo.DDR1 silencing reversed these effects,confirming that DDR1 has a pivotal role in breast cancer recurrence.Conclusion:DNMT3A and METTL7A were shown to cooperatively regulate DDR1 via DNA/m6A methylation,which drives Tregmediated immune suppression and recurrence.This study provided novel insights and therapeutic targets for breast cancer prognosis and treatment.
