Objectives:Pancreatic cancer(PC)is characterized by poor prognosis due to its limited treatment choices and delayed detection.S100A14 has been implicated in tumor progression,yet its regulatory hierarchy and functiona...Objectives:Pancreatic cancer(PC)is characterized by poor prognosis due to its limited treatment choices and delayed detection.S100A14 has been implicated in tumor progression,yet its regulatory hierarchy and functional interplay in PC remain unclear.This study aimed to define the role of S100A14 in PC progression.Methods:Integrated bioinformatic analyses of TCGA-PAAD and GSE22780 datasets identified candidate hub genes.Prognostic relevance was assessed via Kaplan-Meier and ROC analyses.Functional experiments were performed in PANC-1 and BxPC-3 cells,including qRT-PCR,CCK-8 assay,Western blotting,Transwell assay,and apoptosis assay.Co-immunoprecipitation(Co-IP)was used to verify S100A14-S100A16 interaction.CHX chase and dual-luciferase assays were employed to assess protein stability and transcriptional activity.Results:S100A14 was markedly upregulated in PC tissues and cell lines and identified as a key prognostic gene.Silencing S100A14 suppressed EMT,proliferation,invasion,and migration,while reversing S100A16-mediated p53 inhibition and enhancing apoptosis.Mechanistically,Co-IP assay confirmed the protein interaction between S100A14 and S100A16;S100A14 stabilized S100A16 protein through post-translational modification without transcriptional regulation;the S100A14/S100A16 axis reduced p53 protein stability and inhibited its transcriptional activity as well as the downstream p21 expression.Critically,knockdown of S100A14 abrogated the pro-metastatic phenotype of cancer cells.Conclusion:This study identifies S100A14 promotes PC progression by stabilizing S100A16 and suppressing the tumor-suppressive p53/p21 pathway;knockdown of S100A14 can reverse the above effects,restore p53 function,and enhance cancer cell apoptosis.Targeting the S100A14/S100A16/p53 regulatory axis could represent a promising therapeutic approach for PC.展开更多
目的探讨溶质载体家族39成员14(solute carrier family 39 member 14,SLC39A14)对铁过载诱导HepG2细胞胰岛素抵抗的影响及机制。方法体外培养HepG2细胞,加入不同浓度的柠檬酸铁铵(ferric ammonium citrate,FAC)通过CCK-8检测细胞活力、...目的探讨溶质载体家族39成员14(solute carrier family 39 member 14,SLC39A14)对铁过载诱导HepG2细胞胰岛素抵抗的影响及机制。方法体外培养HepG2细胞,加入不同浓度的柠檬酸铁铵(ferric ammonium citrate,FAC)通过CCK-8检测细胞活力、钙黄绿素(Calcein-AM)检测不稳定铁池(labile iron loop,LIP)、透射电镜观察线粒体、细胞葡萄糖消耗量检测胰岛素抵抗情况、以及SLC39A14蛋白和mRNA的表达;随后加入铁死亡抑制剂(Ferrostatin-1,Fer-1)后检测活性氧(reactive oxygen species,ROS)、丙二醛(malondialdehyde,MDA)和谷胱甘肽(glutathione,GSH);最后使用小干扰RNA(siRNA)技术敲低SLC39A14的表达,观察氧化应激及胰岛素抵抗指标是否变化。结果FAC干预后细胞活力下降、LIP水平增加、细胞葡萄糖消耗量减少,同时上调SLC39A14蛋白和mRNA的表达。与FAC组比较,加入铁死亡抑制剂联合处理后,通过减少ROS和MDA水平、增加GSH含量等方式改善细胞内氧化应激水平,下调SLC39A14蛋白和mRNA的表达,增加葡萄糖消耗量,进而缓解胰岛素抵抗的发生。敲低SLC39A14后,FAC+SLC39A14敲低组HepG2细胞氧化应激和胰岛素抵抗水平较FAC组进一步改善。结论FAC引起HepG2细胞胰岛素抵抗的发生,机制可能与激活SLC39A14有关。展开更多
BACKGROUND Hepatocellular carcinoma(HCC)is characterized by high morbidity and mortality owing to its mechanistic complexity and individual heterogeneity,making early diagnosis challenging.The role of SLC39A14,a bioma...BACKGROUND Hepatocellular carcinoma(HCC)is characterized by high morbidity and mortality owing to its mechanistic complexity and individual heterogeneity,making early diagnosis challenging.The role of SLC39A14,a biomarker for multiple tumors,in HCC remains to be elucidated.AIM To determine the tumor-suppressive role of SLC39A14 in HCC and its underlying molecular mechanisms.METHODS The expression pattern of SLC39A14 in HCC was evaluated using quantitative reverse transcriptase PCR and western blotting,and its association with tumor malignancy was further validated in C57BL/6J mouse HCC model.A series of functional assays,including cell counting kit-8,colony formation,apoptosis,scratch,and invasion tests,were conducted to assess how alterations in the SLC39A14 expression affect the oncological behavior of HCC cells.In addition,bioinformatics analysis was performed to investigate the potential regulatory mechanism underlying SLC39A14 expression in HCC.RESULTS The protein expression and RNA abundance of SLC39A14 in human HCC tissues were significantly lower than those in paracancerous tissues(P<0.01).In HCC model mice,the SLC39A14 expression decreased gradually as the disease progressed.The overexpression of SLC39A14 significantly inhibited the proliferation,migration,and invasion of Huh7 HCC cells while promoting their apoptosis.The knockdown of SLC39A14 exerted the opposite effect.Bioinformatics analysis suggested the involvement of the transcription factor STAT3 in regulating the SLC39A14 expression.CONCLUSION SLC39A14 dysregulation promotes HCC progression through BCL-2/BAX/Caspase-3 apoptotic axis and epithelial–mesenchymal transition transformation axis,suggesting potential therapeutic targets.展开更多
基金supported by the Yunnan Province Liu Liang Expert Workstation(No.202305AF150148)Famous Doctor Projects of Yunnan Province(No.XDYC-MY-2022-0032)+1 种基金Yunnan Health Training Project of High Level Talents(No.L-2024029)Innovation Team Special Program of Yunnan(No.202505AS350004).
文摘Objectives:Pancreatic cancer(PC)is characterized by poor prognosis due to its limited treatment choices and delayed detection.S100A14 has been implicated in tumor progression,yet its regulatory hierarchy and functional interplay in PC remain unclear.This study aimed to define the role of S100A14 in PC progression.Methods:Integrated bioinformatic analyses of TCGA-PAAD and GSE22780 datasets identified candidate hub genes.Prognostic relevance was assessed via Kaplan-Meier and ROC analyses.Functional experiments were performed in PANC-1 and BxPC-3 cells,including qRT-PCR,CCK-8 assay,Western blotting,Transwell assay,and apoptosis assay.Co-immunoprecipitation(Co-IP)was used to verify S100A14-S100A16 interaction.CHX chase and dual-luciferase assays were employed to assess protein stability and transcriptional activity.Results:S100A14 was markedly upregulated in PC tissues and cell lines and identified as a key prognostic gene.Silencing S100A14 suppressed EMT,proliferation,invasion,and migration,while reversing S100A16-mediated p53 inhibition and enhancing apoptosis.Mechanistically,Co-IP assay confirmed the protein interaction between S100A14 and S100A16;S100A14 stabilized S100A16 protein through post-translational modification without transcriptional regulation;the S100A14/S100A16 axis reduced p53 protein stability and inhibited its transcriptional activity as well as the downstream p21 expression.Critically,knockdown of S100A14 abrogated the pro-metastatic phenotype of cancer cells.Conclusion:This study identifies S100A14 promotes PC progression by stabilizing S100A16 and suppressing the tumor-suppressive p53/p21 pathway;knockdown of S100A14 can reverse the above effects,restore p53 function,and enhance cancer cell apoptosis.Targeting the S100A14/S100A16/p53 regulatory axis could represent a promising therapeutic approach for PC.
文摘目的探讨溶质载体家族39成员14(solute carrier family 39 member 14,SLC39A14)对铁过载诱导HepG2细胞胰岛素抵抗的影响及机制。方法体外培养HepG2细胞,加入不同浓度的柠檬酸铁铵(ferric ammonium citrate,FAC)通过CCK-8检测细胞活力、钙黄绿素(Calcein-AM)检测不稳定铁池(labile iron loop,LIP)、透射电镜观察线粒体、细胞葡萄糖消耗量检测胰岛素抵抗情况、以及SLC39A14蛋白和mRNA的表达;随后加入铁死亡抑制剂(Ferrostatin-1,Fer-1)后检测活性氧(reactive oxygen species,ROS)、丙二醛(malondialdehyde,MDA)和谷胱甘肽(glutathione,GSH);最后使用小干扰RNA(siRNA)技术敲低SLC39A14的表达,观察氧化应激及胰岛素抵抗指标是否变化。结果FAC干预后细胞活力下降、LIP水平增加、细胞葡萄糖消耗量减少,同时上调SLC39A14蛋白和mRNA的表达。与FAC组比较,加入铁死亡抑制剂联合处理后,通过减少ROS和MDA水平、增加GSH含量等方式改善细胞内氧化应激水平,下调SLC39A14蛋白和mRNA的表达,增加葡萄糖消耗量,进而缓解胰岛素抵抗的发生。敲低SLC39A14后,FAC+SLC39A14敲低组HepG2细胞氧化应激和胰岛素抵抗水平较FAC组进一步改善。结论FAC引起HepG2细胞胰岛素抵抗的发生,机制可能与激活SLC39A14有关。
基金Supported by Postgraduate Research&Practice Innovation Program of Jiangsu Province,No.SJCX23-0857.
文摘BACKGROUND Hepatocellular carcinoma(HCC)is characterized by high morbidity and mortality owing to its mechanistic complexity and individual heterogeneity,making early diagnosis challenging.The role of SLC39A14,a biomarker for multiple tumors,in HCC remains to be elucidated.AIM To determine the tumor-suppressive role of SLC39A14 in HCC and its underlying molecular mechanisms.METHODS The expression pattern of SLC39A14 in HCC was evaluated using quantitative reverse transcriptase PCR and western blotting,and its association with tumor malignancy was further validated in C57BL/6J mouse HCC model.A series of functional assays,including cell counting kit-8,colony formation,apoptosis,scratch,and invasion tests,were conducted to assess how alterations in the SLC39A14 expression affect the oncological behavior of HCC cells.In addition,bioinformatics analysis was performed to investigate the potential regulatory mechanism underlying SLC39A14 expression in HCC.RESULTS The protein expression and RNA abundance of SLC39A14 in human HCC tissues were significantly lower than those in paracancerous tissues(P<0.01).In HCC model mice,the SLC39A14 expression decreased gradually as the disease progressed.The overexpression of SLC39A14 significantly inhibited the proliferation,migration,and invasion of Huh7 HCC cells while promoting their apoptosis.The knockdown of SLC39A14 exerted the opposite effect.Bioinformatics analysis suggested the involvement of the transcription factor STAT3 in regulating the SLC39A14 expression.CONCLUSION SLC39A14 dysregulation promotes HCC progression through BCL-2/BAX/Caspase-3 apoptotic axis and epithelial–mesenchymal transition transformation axis,suggesting potential therapeutic targets.
