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
目的通过生物信息学分析SLC7A5在乳腺癌中的表达及其与临床病理特征的相关性,为乳腺癌的治疗及预后评估提供参考。方法在Timer2.0、癌症基因组图谱(the cancer genome atlas,TCGA)、UALAN在基因表达谱交互式分析(gene expression profil...目的通过生物信息学分析SLC7A5在乳腺癌中的表达及其与临床病理特征的相关性,为乳腺癌的治疗及预后评估提供参考。方法在Timer2.0、癌症基因组图谱(the cancer genome atlas,TCGA)、UALAN在基因表达谱交互式分析(gene expression profiling interactive analysis 2,GEPIA2)和人类蛋白质表达图谱(human protein atlas,HPA)等数据库中获取SLC7A5在乳腺癌中的表达特征。使用Western blot法检测SLC7A5基因在正常乳腺细胞MCF-10A和乳腺癌细胞株MCF-7中的表达情况。使用R软件分析SLC7A5在乳腺癌中的预后价值,在Kaplan-Meier Plotter和基因表达数据库(gene expression omnibus,GEO)中进行双重验证,同时分析其表达与乳腺癌患者不同病理特征之间的相关性。运用R语言分析SLC7A5与肿瘤突变负荷(tumor mutation burden,TMB)及免疫检测点相关基因的相关性,并进行基因集富集分析(gene set enrichment analysis,GSEA),并对分析结果进一步可视化处理。结果SLC7A5在包括乳腺癌在内的多种实体恶性肿瘤中表达升高,高表达SLC7A5的乳腺癌患者总生存期缩短。SLC7A5在乳腺癌中的表达水平与乳腺癌雌激素受体、孕激素受体、淋巴结转移及分子分型相关。TMB结果提示,SLC7A5在乳腺癌中的表达与TMB呈正相关(r=0.33,P<0.001)。免疫检查点分析结果表明,SLC7A5的表达量与程序性死亡受体1(programmed death-1,PD-1)、细胞毒性T淋巴细胞相关蛋白4(cytotoxic T lymphocyte-associated antigen 4,CTLA4)、B和T细胞衰减因子(B and T lymphocyte attenuator,BTLA)等呈正相关。GSEA富集分析结果显示SLC7A5主要与干扰素γ反应相关通路相关。结论SLC7A5在乳腺癌中表达上调,其表达水平与不良预后相关,可能通过参与干扰素γ反应相关通路促进免疫检查点的表达影响乳腺癌的进展,其或可作为为乳腺癌患者预后评估和指导治疗的生物学标志物。展开更多
Objectives:Triple-negative breast cancer(TNBC)is the breast cancer subtype with the poorest prognosis.This study aimed to elucidate the molecular pathways through which isoliquiritigenin(ISL),a natural chalcone compou...Objectives:Triple-negative breast cancer(TNBC)is the breast cancer subtype with the poorest prognosis.This study aimed to elucidate the molecular pathways through which isoliquiritigenin(ISL),a natural chalcone compound derived fromlicorice and other plant roots,targets interferon regulatory factor 5(IRF5)in TNBC.Methods:TNBC cell lines were cultured and subjected to IRF5 knockdown using short hairpin RNA.Cell proliferation was assessed by cell counting kit-8(CCK-8)assay and colony formation assays.Western blotting and quantitative reverse transcription polymerase chain reaction(RT-PCR)were employed to measure expression levels of IRF5,solute carrier family 7 member 5(SLC7A5),and indoleamine 2,3-dioxygenase 1(IDO1).Intracellular tryptophan and its metabolites were quantified using commercially available assay kits and high-performance liquid chromatography(HPLC).TNBC cells were treated with various concentrations of ISL to evaluate its effects on proliferation and tryptophanmetabolism.Results:IRF5 was highly expressed in TNBC cell lines.Silencing IRF5 significantly inhibited cellular proliferation and growth.Knockdown of IRF5 reduced the expression of SLC7A5 and IDO1,leading to decreased intracellular levels of tryptophan and its metabolites.ISL markedly suppressed TNBC cell proliferation and disrupted tryptophan metabolism in tumor cells.Conclusion:ISL may inhibit TNBC progression by downregulating IRF5 and interfering with SLC7A5/IDO1-mediated tryptophan metabolic reprogramming,suggesting a potential therapeutic mechanism for TNBC treatment.展开更多
Insects produce silk to form cocoons,nests,and webs,which are important for their survival and reproduction.However,little is known about the molecular mecha-nism of silk protein synthesis at the translation level.The...Insects produce silk to form cocoons,nests,and webs,which are important for their survival and reproduction.However,little is known about the molecular mecha-nism of silk protein synthesis at the translation level.The solute carrier family 7(SLC7)genes are involved in activating the target of rapamycin complex 1(TORC1)signaling pathway and protein translation process,but the physiological roles of SLC7 genes in silk-producing insects have not been reported.Here,we found that amino acid signaling regulates silk protein synthesis and larval development via the L-type amino acid trans-porter 1(LAT1;also known as SLC7A5)in Bombyx mori.A total of 12 SLC7 homologs were identified in the silkworm genome,among which BmSLC7A5 was found to be a silk gland-enriched gene and may be involved in leucine transport.Bioinformatics analy-sis indicated that SLC7A5 displays high homology and a close phylogenetic relationship in silk-producing insects.Subsequently,we found that leucine treatment significantly in-creased silk protein synthesis by improving the transcription and protein levels of silk genes.Furthermore,systemic and silk gland-specific knockout of BmSLC7A5 led to de-creased silk protein synthesis by inhibiting TORC1 signaling,and somatic mutation also resulted in arrested development from the 5th instar to the early pupal stage.Altogether,our study reveals that BmSLC7A5 is involved in regulating silk protein synthesis and larval development by affecting the TORC1 signaling pathway,which provides a new strategy and target for improving silk yield.展开更多
目的探讨实时荧光定量PCR SYBR Green I探针技术用于骨髓增生异常综合征(MDS)患者基因检测的可行性。方法运用实时荧光定量PCR SYBR Green I技术,检测21例MDS患者的SLC7A5基因表达情况,并以IDA、ITP、增贫及其他贫血19例为对照组。结果...目的探讨实时荧光定量PCR SYBR Green I探针技术用于骨髓增生异常综合征(MDS)患者基因检测的可行性。方法运用实时荧光定量PCR SYBR Green I技术,检测21例MDS患者的SLC7A5基因表达情况,并以IDA、ITP、增贫及其他贫血19例为对照组。结果实验组和对照组SLC7A5基因的表达无明显的统计学差异。结论real-time PCR技术检测目的基因的表达是非常方便、快捷、准确的方法。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。
文摘目的通过生物信息学分析SLC7A5在乳腺癌中的表达及其与临床病理特征的相关性,为乳腺癌的治疗及预后评估提供参考。方法在Timer2.0、癌症基因组图谱(the cancer genome atlas,TCGA)、UALAN在基因表达谱交互式分析(gene expression profiling interactive analysis 2,GEPIA2)和人类蛋白质表达图谱(human protein atlas,HPA)等数据库中获取SLC7A5在乳腺癌中的表达特征。使用Western blot法检测SLC7A5基因在正常乳腺细胞MCF-10A和乳腺癌细胞株MCF-7中的表达情况。使用R软件分析SLC7A5在乳腺癌中的预后价值,在Kaplan-Meier Plotter和基因表达数据库(gene expression omnibus,GEO)中进行双重验证,同时分析其表达与乳腺癌患者不同病理特征之间的相关性。运用R语言分析SLC7A5与肿瘤突变负荷(tumor mutation burden,TMB)及免疫检测点相关基因的相关性,并进行基因集富集分析(gene set enrichment analysis,GSEA),并对分析结果进一步可视化处理。结果SLC7A5在包括乳腺癌在内的多种实体恶性肿瘤中表达升高,高表达SLC7A5的乳腺癌患者总生存期缩短。SLC7A5在乳腺癌中的表达水平与乳腺癌雌激素受体、孕激素受体、淋巴结转移及分子分型相关。TMB结果提示,SLC7A5在乳腺癌中的表达与TMB呈正相关(r=0.33,P<0.001)。免疫检查点分析结果表明,SLC7A5的表达量与程序性死亡受体1(programmed death-1,PD-1)、细胞毒性T淋巴细胞相关蛋白4(cytotoxic T lymphocyte-associated antigen 4,CTLA4)、B和T细胞衰减因子(B and T lymphocyte attenuator,BTLA)等呈正相关。GSEA富集分析结果显示SLC7A5主要与干扰素γ反应相关通路相关。结论SLC7A5在乳腺癌中表达上调,其表达水平与不良预后相关,可能通过参与干扰素γ反应相关通路促进免疫检查点的表达影响乳腺癌的进展,其或可作为为乳腺癌患者预后评估和指导治疗的生物学标志物。
基金supported by the Science and Technology Program of Guangzhou(No.202103000020).
文摘Objectives:Triple-negative breast cancer(TNBC)is the breast cancer subtype with the poorest prognosis.This study aimed to elucidate the molecular pathways through which isoliquiritigenin(ISL),a natural chalcone compound derived fromlicorice and other plant roots,targets interferon regulatory factor 5(IRF5)in TNBC.Methods:TNBC cell lines were cultured and subjected to IRF5 knockdown using short hairpin RNA.Cell proliferation was assessed by cell counting kit-8(CCK-8)assay and colony formation assays.Western blotting and quantitative reverse transcription polymerase chain reaction(RT-PCR)were employed to measure expression levels of IRF5,solute carrier family 7 member 5(SLC7A5),and indoleamine 2,3-dioxygenase 1(IDO1).Intracellular tryptophan and its metabolites were quantified using commercially available assay kits and high-performance liquid chromatography(HPLC).TNBC cells were treated with various concentrations of ISL to evaluate its effects on proliferation and tryptophanmetabolism.Results:IRF5 was highly expressed in TNBC cell lines.Silencing IRF5 significantly inhibited cellular proliferation and growth.Knockdown of IRF5 reduced the expression of SLC7A5 and IDO1,leading to decreased intracellular levels of tryptophan and its metabolites.ISL markedly suppressed TNBC cell proliferation and disrupted tryptophan metabolism in tumor cells.Conclusion:ISL may inhibit TNBC progression by downregulating IRF5 and interfering with SLC7A5/IDO1-mediated tryptophan metabolic reprogramming,suggesting a potential therapeutic mechanism for TNBC treatment.
基金Thiswork was supported bygrants fromthe NationalNaturalScience FoundationofChina(31772532)the China Postdoctoral Science Foundation(2022MD713704)the Chongqing Science and Technology Bureau(cstc2021ljcyj-bshX0222 and jbky20210004).
文摘Insects produce silk to form cocoons,nests,and webs,which are important for their survival and reproduction.However,little is known about the molecular mecha-nism of silk protein synthesis at the translation level.The solute carrier family 7(SLC7)genes are involved in activating the target of rapamycin complex 1(TORC1)signaling pathway and protein translation process,but the physiological roles of SLC7 genes in silk-producing insects have not been reported.Here,we found that amino acid signaling regulates silk protein synthesis and larval development via the L-type amino acid trans-porter 1(LAT1;also known as SLC7A5)in Bombyx mori.A total of 12 SLC7 homologs were identified in the silkworm genome,among which BmSLC7A5 was found to be a silk gland-enriched gene and may be involved in leucine transport.Bioinformatics analy-sis indicated that SLC7A5 displays high homology and a close phylogenetic relationship in silk-producing insects.Subsequently,we found that leucine treatment significantly in-creased silk protein synthesis by improving the transcription and protein levels of silk genes.Furthermore,systemic and silk gland-specific knockout of BmSLC7A5 led to de-creased silk protein synthesis by inhibiting TORC1 signaling,and somatic mutation also resulted in arrested development from the 5th instar to the early pupal stage.Altogether,our study reveals that BmSLC7A5 is involved in regulating silk protein synthesis and larval development by affecting the TORC1 signaling pathway,which provides a new strategy and target for improving silk yield.
文摘目的探讨实时荧光定量PCR SYBR Green I探针技术用于骨髓增生异常综合征(MDS)患者基因检测的可行性。方法运用实时荧光定量PCR SYBR Green I技术,检测21例MDS患者的SLC7A5基因表达情况,并以IDA、ITP、增贫及其他贫血19例为对照组。结果实验组和对照组SLC7A5基因的表达无明显的统计学差异。结论real-time PCR技术检测目的基因的表达是非常方便、快捷、准确的方法。
