The speckle-type POZ protein (SPOP) is a tumor suppressor in prostate cancer (PCa). SPOP somatic mutations have been reported in up to 15% of PCa of those of European descent. However, the genetic roles of SPOP in...The speckle-type POZ protein (SPOP) is a tumor suppressor in prostate cancer (PCa). SPOP somatic mutations have been reported in up to 15% of PCa of those of European descent. However, the genetic roles of SPOP in African American (AA)-PCa are currently unknown. We sequenced the SPOP gene to identify somatic mutations in 49 AA prostate tumors and identified three missense mutations (p.Y87C, p.F102S, and p.G111E) in five AA prostate tumors (10%) and one synonymous variant (p.11061) in one tumor. Intriguingly, all of mutations and variants clustered in exon six, and all of the mutations altered conserved amino acids. Moreover, two mutations (p.F102S and p.G111E) have only been identified in AA-PCa to date. Quantitative real-time polymerase chain reaction analysis showed a lower level of SPOP expression in tumors carrying SPOP mutations than their matched normal prostate tissues. In addition, SPOP mutations and novel variants were detected in 5 of 27 aggressive PCa and one of 22 less aggressive PCa (P 〈 0.05). Further studies with increased sample size are needed to validate the clinicopathological significance of these SPOP mutations in AA-PCa.展开更多
目的:观察Rho激酶抑制剂法舒地尔(Fasudil)对人高侵袭潜能HCC细胞株3(human high metastatic liver cancer cells 3,HCCLM3)侵袭转移的影响,并且探讨其作用的机制。方法:应用100 mol/L Fasudil作用于HCCLM3细胞,采用肌动蛋白微丝荧光染...目的:观察Rho激酶抑制剂法舒地尔(Fasudil)对人高侵袭潜能HCC细胞株3(human high metastatic liver cancer cells 3,HCCLM3)侵袭转移的影响,并且探讨其作用的机制。方法:应用100 mol/L Fasudil作用于HCCLM3细胞,采用肌动蛋白微丝荧光染色和侵袭小室实验观察HCCLM3细胞的运动侵袭能力。HCCLM3细胞经过处理后分为阴性对照组、Fasudil作用组、BTBD7干扰组,通过Western印迹检测BTB/POZ结构域蛋白7(BR-C,ttk and bab/pox virus domain containing 7,BTBD7)、Ras同系物家族成员C(ras homolog family member C,Rho C)、Rho关联卷曲螺旋蛋白激酶2(Rhoassociated,coiled-coil containing protein kinase 2,ROCK2)、MMP2和MMP9蛋白表达水平,酶谱分析法检测MMP2和MMP9活性水平。BTBD7干扰组作为阳性对照。结果:Fasudil处理后HCCLM3侵袭运动能力下降,BTBD7,Rho C,ROCK2蛋白表达下调,MMP2和MMP9活性降低,与阴性对照组比较差异有统计学意义(均P<0.01)。结论:Fasudil具有干预BTBD7-ROCK2信号通路、抑制HCC侵袭转移的重要作用。展开更多
在胚胎发育中发挥重要作用的许多基因,其异常表达或突变常与疾病密切相关,斑点型BTB/POZ蛋白(speckle type BTB/POZ protein,SPOP)是其中之一。SPOP是E3泛素连接酶接头蛋白质,主要由MATH、BTB和BACK结构域构成,其功能正常发挥依赖于多...在胚胎发育中发挥重要作用的许多基因,其异常表达或突变常与疾病密切相关,斑点型BTB/POZ蛋白(speckle type BTB/POZ protein,SPOP)是其中之一。SPOP是E3泛素连接酶接头蛋白质,主要由MATH、BTB和BACK结构域构成,其功能正常发挥依赖于多个结构域各自不同的作用。SPOP主要通过泛素-蛋白酶体途径促进其靶蛋白质的降解来发挥作用。目前发现,SPOP底物蛋白质有30多种,其中大部分与前列腺癌、子宫内膜癌和肾癌的发生发展相关。SPOP在机体发育过程中也发挥重要作用,缺失或者突变SPOP导致小鼠出生后死亡。SPOP新生突变导致儿童神经发育障碍。SPOP调控机体发育也主要通过调控靶蛋白质降解来实现,包括作用于Gli2/3、PDX1、NANOG和SENP7等靶蛋白质来调控神经、骨骼和胰腺的发育以及衰老等过程。另有研究发现,SPOP与靶蛋白质会共定位到核斑(nuclear speckles)等无膜细胞器中,促进靶蛋白质的泛素化降解,并且SPOP寡聚体的形成以及其与底物多价互作引发的液-液相分离(liquid-liquid phase separation,LLPS)也在其中发挥了重要作用。SPOP寡聚化缺陷的BTB突变或BACK突变,可导致SPOP无法发生液-液相分离并定位于无膜细胞器。本文综合了最新研究进展,详细探讨了SPOP在机体发育过程中的重要作用。展开更多
目的:探讨BTB/POZ结构域蛋白7(BTBD7)假基因1(BTBD7P1)在肝细胞癌(HCC)中的表达及功能。方法:检测106例配对的HCC组织与癌旁组织标本中BTBD7P1 m RNA的表达,分析BTBD7P1m RNA表达与HCC患者临床病理特征及预后的关系。用BTBD7P1过表达慢...目的:探讨BTB/POZ结构域蛋白7(BTBD7)假基因1(BTBD7P1)在肝细胞癌(HCC)中的表达及功能。方法:检测106例配对的HCC组织与癌旁组织标本中BTBD7P1 m RNA的表达,分析BTBD7P1m RNA表达与HCC患者临床病理特征及预后的关系。用BTBD7P1过表达慢病毒载体转染HCC细胞系Bel7404后,检测细胞增殖率以及BTBD7 m RNA与蛋白的表达。结果:HCC组织中BTBD7P1相对表达量明显低于癌旁组织为(0.71 vs.2.14,P<0.05);BTBD7P1m RNA低表达与肿瘤大小、卫星灶、分化程度、静脉血管侵犯、出血坏死、HCC分期明显有关(均P<0.05);BTBD7P1 m RNA低表达患者的1、3、5年总体生存率及无瘤生存率均明显低于BTBD7P1m RNA高表达患者(均P<0.05)。与转染空载体质粒的对照组Bel7404细胞比较,转染BTBD7P1过表达慢病毒载体的Bel7404细胞,细胞增殖能力明显减低,BTBD7 m RNA表达明显下调(均P<0.05),但BTBD7蛋白表达无明显变化(P>0.05)。结论:BTBD7P1可能在m RNA水平对亲本基因BTBD7表达进行调控,从而参与了HCC发生与发展。展开更多
文摘The speckle-type POZ protein (SPOP) is a tumor suppressor in prostate cancer (PCa). SPOP somatic mutations have been reported in up to 15% of PCa of those of European descent. However, the genetic roles of SPOP in African American (AA)-PCa are currently unknown. We sequenced the SPOP gene to identify somatic mutations in 49 AA prostate tumors and identified three missense mutations (p.Y87C, p.F102S, and p.G111E) in five AA prostate tumors (10%) and one synonymous variant (p.11061) in one tumor. Intriguingly, all of mutations and variants clustered in exon six, and all of the mutations altered conserved amino acids. Moreover, two mutations (p.F102S and p.G111E) have only been identified in AA-PCa to date. Quantitative real-time polymerase chain reaction analysis showed a lower level of SPOP expression in tumors carrying SPOP mutations than their matched normal prostate tissues. In addition, SPOP mutations and novel variants were detected in 5 of 27 aggressive PCa and one of 22 less aggressive PCa (P 〈 0.05). Further studies with increased sample size are needed to validate the clinicopathological significance of these SPOP mutations in AA-PCa.
文摘目的:观察Rho激酶抑制剂法舒地尔(Fasudil)对人高侵袭潜能HCC细胞株3(human high metastatic liver cancer cells 3,HCCLM3)侵袭转移的影响,并且探讨其作用的机制。方法:应用100 mol/L Fasudil作用于HCCLM3细胞,采用肌动蛋白微丝荧光染色和侵袭小室实验观察HCCLM3细胞的运动侵袭能力。HCCLM3细胞经过处理后分为阴性对照组、Fasudil作用组、BTBD7干扰组,通过Western印迹检测BTB/POZ结构域蛋白7(BR-C,ttk and bab/pox virus domain containing 7,BTBD7)、Ras同系物家族成员C(ras homolog family member C,Rho C)、Rho关联卷曲螺旋蛋白激酶2(Rhoassociated,coiled-coil containing protein kinase 2,ROCK2)、MMP2和MMP9蛋白表达水平,酶谱分析法检测MMP2和MMP9活性水平。BTBD7干扰组作为阳性对照。结果:Fasudil处理后HCCLM3侵袭运动能力下降,BTBD7,Rho C,ROCK2蛋白表达下调,MMP2和MMP9活性降低,与阴性对照组比较差异有统计学意义(均P<0.01)。结论:Fasudil具有干预BTBD7-ROCK2信号通路、抑制HCC侵袭转移的重要作用。
文摘目的:探讨BTB/POZ结构域蛋白7(BTBD7)假基因1(BTBD7P1)在肝细胞癌(HCC)中的表达及功能。方法:检测106例配对的HCC组织与癌旁组织标本中BTBD7P1 m RNA的表达,分析BTBD7P1m RNA表达与HCC患者临床病理特征及预后的关系。用BTBD7P1过表达慢病毒载体转染HCC细胞系Bel7404后,检测细胞增殖率以及BTBD7 m RNA与蛋白的表达。结果:HCC组织中BTBD7P1相对表达量明显低于癌旁组织为(0.71 vs.2.14,P<0.05);BTBD7P1m RNA低表达与肿瘤大小、卫星灶、分化程度、静脉血管侵犯、出血坏死、HCC分期明显有关(均P<0.05);BTBD7P1 m RNA低表达患者的1、3、5年总体生存率及无瘤生存率均明显低于BTBD7P1m RNA高表达患者(均P<0.05)。与转染空载体质粒的对照组Bel7404细胞比较,转染BTBD7P1过表达慢病毒载体的Bel7404细胞,细胞增殖能力明显减低,BTBD7 m RNA表达明显下调(均P<0.05),但BTBD7蛋白表达无明显变化(P>0.05)。结论:BTBD7P1可能在m RNA水平对亲本基因BTBD7表达进行调控,从而参与了HCC发生与发展。