背景与目的:越来越多的研究表明,序列相似性为83的家族成员A(family with sequence similarity 83 member A,FAM83A)与肿瘤的侵袭和转移相关,但FAM83A在肺腺癌中的作用尚不清楚。探讨FAM83A在肺腺癌组织中的表达情况,与肺腺癌患者预后...背景与目的:越来越多的研究表明,序列相似性为83的家族成员A(family with sequence similarity 83 member A,FAM83A)与肿瘤的侵袭和转移相关,但FAM83A在肺腺癌中的作用尚不清楚。探讨FAM83A在肺腺癌组织中的表达情况,与肺腺癌患者预后的关系以及对肺腺癌细胞增殖、侵袭转移能力的影响。方法:运用人类蛋白质图谱(Human Protein Atlas,HPA)和基因表达谱动态分析(Gene Expression Profiling Interactive Analysis,GEPIA)数据库分析并选取与肺腺癌预后相关的基因;运用UALCAN和GEPIA数据库研究FAM83A的表达量对肺腺癌生存率的影响;采用UALCAN数据库分析FAM83A在肺腺癌组织中的表达及其与肺腺癌患者的性别、淋巴结的分期和转移的关系;采用蛋白质印迹法(Western blot)检测人正常肺上皮细胞BEAS-2B和肺腺癌细胞A549中FAM83A蛋白的表达情况及FAM83A的转染效率;采用细胞计数试剂盒(cell counting kit-8,CCK-8)检测转染后敲减FAM83A对肺腺癌细胞增殖能力的影响;采用transwell侵袭实验检测FAM83A对肺腺癌细胞侵袭能力的影响。结果:采用HPA、GEPIA和UALCAN数据库分析发现高表达的FAM83A与肺腺癌不良预后相关且FAM83A在肺腺癌组织中高表达,差异有统计学意义(P<0.05);FAM83A的表达与肺腺癌的临床分期和淋巴结转移相关;Western blot检测结果显示,FAM83A在肺腺癌细胞中高表达且敲减质粒转染成功;CCK-8细胞增殖实验结果提示FAM83A可促进细胞的增殖能力(P<0.05);Transwell侵袭实验提示FAM83A促进A549的侵袭能力(P<0.05)。结论:FAM83A在肺腺癌组织和细胞中高表达且促进肺腺癌细胞的增殖、侵袭和转移。展开更多
目的探索序列相似性83家族成员A(family with sequence similarity 83 member A,FAM83A)在结直肠癌的表达情况,和敲低FAM83A对结直肠癌细胞增殖能力的影响及相关机制。方法用免疫组化检测102例结直肠癌患者组织及其癌旁组织标本中FAM83...目的探索序列相似性83家族成员A(family with sequence similarity 83 member A,FAM83A)在结直肠癌的表达情况,和敲低FAM83A对结直肠癌细胞增殖能力的影响及相关机制。方法用免疫组化检测102例结直肠癌患者组织及其癌旁组织标本中FAM83A的表达情况。将HCT116细胞分为实验组和对照组,实验组细胞转染FAM83A-siRNA质粒,对照组细胞转染MOCK-siRNA质粒,荧光定量PCR检测各组细胞FAM83AmRNA含量,蛋白质印迹法检测各组细胞的FAM83A、PI3K、p-AKT、p-mTOR表达情况,CCK8实验和克隆形成实验检测各组细胞的增殖能力。结果果结直肠癌患者组织FAM83A阳性率为88.23%(90/102),癌旁组织中表达率为10.78%(11/102),FAM83A在结直肠癌组织中的表达率显著高于癌旁组织,差异具有统计学意义(P<0.001)。转染siRNA后,实验组和对照组HCT116细胞中FAM83AmRNA表达量分别为1.23±0.20和0.43±0.12,FAM83A蛋白表达量分别为1.19±0.11和0.23±0.08,PI3K的表达量分别为1.21±0.17、0.28±0.09,p-AKT的表达量分别为1.35±0.23、0.57±0.18,p-mTOR表达量分别为1.48±0.20、1.05±0.14,PI3K、p-AKT、p-mTOR表达均下调(均P<0.05)。实验组和对照组HCT116细胞CCK8实验120h的吸光度分别为1.09±0.22和2.21±0.27实验组和对照组HCT116细胞的克隆形成率分别为21.6%±2.4%和62.7%±4.1%,实验组细胞增殖能力明显下降(P<0.05)。结论FAM83A在结直肠癌组织中表达明显升高,可能与结直肠癌的恶性程度相关;且FAM83A通过PI3K/AKT/mTOR信号通路影响结直肠癌细胞的增殖能力。展开更多
Abnormal activation of Wnt/β-catenin-mediated transcription is closely associated with the malignancy of pancreatic cancer.Family with sequence similarity 83 member A(FAM83A)was shown recently to have oncogenic effec...Abnormal activation of Wnt/β-catenin-mediated transcription is closely associated with the malignancy of pancreatic cancer.Family with sequence similarity 83 member A(FAM83A)was shown recently to have oncogenic effects in a variety of cancer types,but the biological roles and molecular mechanisms of FAM83A in pancreatic cancer need further investigation.Here,we newly discovered that FAM83A binds directly toβ-catenin and inhibits the assembly of the cytoplasmic destruction complex thus inhibiting the subsequent phosphorylation and degradation.FAM83A is mainly phosphorylated by the SRC non-receptor kinase family member BLK(B-lymphoid tyrosine kinase)at tyrosine 138 residue within the DUF1669 domain that mediates the FAM83A-β-catenin interaction.Moreover,FAM83A tyrosine 138 phosphorylation enhances oncogenic Wnt/β-catenin-mediated transcription through promotingβ-catenin-TCF4 interaction and showed an elevated nucleus translocation,which inhibits the recruitment of histone deacetylases by TCF4.We also showed that FAM83A is a direct downstream target of Wnt/β-catenin signaling and correlates with the levels of Wnt target genes in human clinical pancreatic cancer tissues.Notably,the inhibitory peptides that target the FAM83A-β-catenin interaction significantly suppressed pancreatic cancer growth and metastasis in vitro and in vivo.Our results revealed that blocking the FAM83A cascade signaling defines a therapeutic target in human pancreatic cancer.展开更多
文摘目的探索序列相似性83家族成员A(family with sequence similarity 83 member A,FAM83A)在结直肠癌的表达情况,和敲低FAM83A对结直肠癌细胞增殖能力的影响及相关机制。方法用免疫组化检测102例结直肠癌患者组织及其癌旁组织标本中FAM83A的表达情况。将HCT116细胞分为实验组和对照组,实验组细胞转染FAM83A-siRNA质粒,对照组细胞转染MOCK-siRNA质粒,荧光定量PCR检测各组细胞FAM83AmRNA含量,蛋白质印迹法检测各组细胞的FAM83A、PI3K、p-AKT、p-mTOR表达情况,CCK8实验和克隆形成实验检测各组细胞的增殖能力。结果果结直肠癌患者组织FAM83A阳性率为88.23%(90/102),癌旁组织中表达率为10.78%(11/102),FAM83A在结直肠癌组织中的表达率显著高于癌旁组织,差异具有统计学意义(P<0.001)。转染siRNA后,实验组和对照组HCT116细胞中FAM83AmRNA表达量分别为1.23±0.20和0.43±0.12,FAM83A蛋白表达量分别为1.19±0.11和0.23±0.08,PI3K的表达量分别为1.21±0.17、0.28±0.09,p-AKT的表达量分别为1.35±0.23、0.57±0.18,p-mTOR表达量分别为1.48±0.20、1.05±0.14,PI3K、p-AKT、p-mTOR表达均下调(均P<0.05)。实验组和对照组HCT116细胞CCK8实验120h的吸光度分别为1.09±0.22和2.21±0.27实验组和对照组HCT116细胞的克隆形成率分别为21.6%±2.4%和62.7%±4.1%,实验组细胞增殖能力明显下降(P<0.05)。结论FAM83A在结直肠癌组织中表达明显升高,可能与结直肠癌的恶性程度相关;且FAM83A通过PI3K/AKT/mTOR信号通路影响结直肠癌细胞的增殖能力。
基金We thank the Hubei University of Technology for the research equipment and technical support for this research.This work was supported by the National Natural Science Foundation of China(32070726 and 82273970 to J.F.T.,32270768 to C.F.Z.,31871176 to X.Z.C.,82173043 to W.Z.T.)Wuhan Science and Technology Project(2019020701011475 to J.F.T.,2022020801020272 to C.F.Z.)+1 种基金National Natural Science Foundation of Hubei(2020CFA073 to J.F.T.,2022EHB038 to C.F.Z.)Doctoral Start-up Foundation of Hubei University of Technology(BSQD2020035 to C.F.Z.).
文摘Abnormal activation of Wnt/β-catenin-mediated transcription is closely associated with the malignancy of pancreatic cancer.Family with sequence similarity 83 member A(FAM83A)was shown recently to have oncogenic effects in a variety of cancer types,but the biological roles and molecular mechanisms of FAM83A in pancreatic cancer need further investigation.Here,we newly discovered that FAM83A binds directly toβ-catenin and inhibits the assembly of the cytoplasmic destruction complex thus inhibiting the subsequent phosphorylation and degradation.FAM83A is mainly phosphorylated by the SRC non-receptor kinase family member BLK(B-lymphoid tyrosine kinase)at tyrosine 138 residue within the DUF1669 domain that mediates the FAM83A-β-catenin interaction.Moreover,FAM83A tyrosine 138 phosphorylation enhances oncogenic Wnt/β-catenin-mediated transcription through promotingβ-catenin-TCF4 interaction and showed an elevated nucleus translocation,which inhibits the recruitment of histone deacetylases by TCF4.We also showed that FAM83A is a direct downstream target of Wnt/β-catenin signaling and correlates with the levels of Wnt target genes in human clinical pancreatic cancer tissues.Notably,the inhibitory peptides that target the FAM83A-β-catenin interaction significantly suppressed pancreatic cancer growth and metastasis in vitro and in vivo.Our results revealed that blocking the FAM83A cascade signaling defines a therapeutic target in human pancreatic cancer.
