CN116426637B - Actn2在预测或检测胃癌骨髓转移中的应用 - Google Patents
Actn2在预测或检测胃癌骨髓转移中的应用 Download PDFInfo
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Abstract
本发明公开了ACTN2在预测或检测胃癌骨髓转移中的应用,涉及生物技术领域。本发明证实了ACTN2表达上调与胃癌骨髓转移有关;发现α‑Actinin‑2是丝状伪足形成所必需的;α‑Actinin‑2直接结合肌动蛋白并促进其组装成丝状伪足;α‑Actinin‑2参与了丝状伪足的形成,其通过取代α‑Actinin‑1形成α‑Actinin‑2:α‑Actinin‑4复合物来促进肌动蛋白丝的交联。本发明将ACTN2及α‑Actinin‑2应用于预测或检测胃癌骨髓转移中,为胃癌骨髓转移提供新的潜在治疗靶点,同时也发掘出了ACTN2及α‑Actinin‑2新的医药价值。
Description
技术领域
本发明涉及生物技术领域,具体涉及ACTN2在预测或检测胃癌骨髓转移中的应用。
背景技术
肝、腹膜和肺是胃癌最常见的转移部位,通过尸检发现在骨髓中也存在高达20%的胃癌细胞,这表明骨髓转移可能在晚期胃癌中被低估。骨髓转移已被用于指代广泛的骨髓浸润,从微转移到明显的播散性癌病。具有明显骨髓转移的晚期胃癌经常导致严重复杂的血液学异常,例如弥散性血管内凝血,其表现为侵袭性但可控制的疾病状态。
α-Actinin-2(由ACTN2基因编码)是F-Actin交联蛋白的α-Actinin家族的成员。α-肌动蛋白-2(由ACTN2基因编码)是F-肌动蛋白交联蛋白α-肌动蛋白家族的成员。人类α-肌动蛋白成员有四种:α-肌动蛋白-1、2、3和4。α-肌动蛋白-1和α-肌动蛋白-2主要在心肌、骨骼肌和大脑中表达,而α-肌动蛋白-3主要在骨骼肌中表达。α-肌动蛋白-2包括三个功能结构域:介导与肌动蛋白相互作用的NH2末端区域、由四个光谱蛋白样重复组成的中心区域以及包含EF手的COOH末端。既往的研究报道中,ACTN2基因突变导致扩张型心肌病。然而在肿瘤研究领域,目前尚无有关α-肌动蛋白-2在癌症发展过程中的确切功能的研究报道。
发明内容
本发明的目的在于克服现有技术的不足,提供ACTN2在预测或检测胃癌骨髓转移中的应用。
为实现上述目的,本发明采取的技术方案为:ACTN2在制备预测或检测胃癌骨髓转移的产品中的应用;所述应用为非疾病诊断和治疗的应用。
伴有骨髓转移的胃癌经常导致严重复杂的血液学异常,如弥漫性血管内凝血和微血管病性溶血性贫血,它们构成了高度侵袭性的胃癌亚型。与常见的晚期胃癌相比,高度侵袭性的胃癌的预后非常差,具有特殊的临床和病理特征。本申请发明人根据高度侵袭性胃癌和常见的晚期胃癌之间的转录组差异阐明了骨髓转移的关键机制,并发现编码蛋白α-Actinin-2的ACTN2基因增强了小鼠模型中胃癌细胞的转移能力并诱导了骨髓转移。
作为本发明所述应用的优选实施方式,所述产品包括检测样本中ACTN2的表达量的试剂。
作为本发明所述应用的优选实施方式,所述样本中ACTN2的表达量上调。
作为本发明所述应用的优选实施方式,所述试剂包括通过PCR或免疫检测检测ACTN2表达水平的试剂。
作为本发明所述应用的优选实施方式,所述产品包括芯片或试剂盒。
本发明还提供ACTN2编码的蛋白在制备预测或检测胃癌骨髓转移的产品中的应用。
作为本发明所述应用的优选实施方式,所述蛋白为α-Actinin-2。
α-Actinin-2由ACTN2基因编码。丝状伪足呈很薄(直径0.1-0.3μm)的手指状,富含肌动蛋白的质膜突起,参与许多细胞过程,包括癌细胞迁移和神经元生长锥寻路。丰富的丝状伪足是肿瘤细胞的标志,并且在许多侵袭性癌细胞类型中得到证实。神经元树突上的棘突代表了丝状突起的形态,而α-Actinin-2在增加海马神经元树突突起的长度和密度中起着关键作用。本申请发明人研究发现,α-Actinin-2是丝状伪足形成所必需的;α-Actinin-2直接结合肌动蛋白并促进其组装成丝状伪足;α-Actinin-2参与了丝状伪足的形成,其通过取代α-Actinin-1形成α-Actinin-2:α-Actinin-4复合物来促进肌动蛋白丝的交联。
作为本发明所述应用的优选实施方式,所述产品包括检测样本中α-Actinin-2的表达量的试剂。
作为本发明所述应用的优选实施方式,所述样本中α-Actinin-2的表达量上调。
本发明还提供一种用于预测或检测胃癌骨髓转移的试剂盒,所述试剂盒含有能检测ACTN2或α-Actinin-2表达量的试剂。
本发明的有益效果:本发明提供了ACTN2在预测或检测胃癌骨髓转移中的应用。本发明证实了ACTN2表达上调与胃癌骨髓转移有关;揭示了α-Actinin-2通过丝状伪足形成促进胃癌中骨髓转移的新作用。本发明将ACTN2及α-Actinin-2应用于预测或检测胃癌骨髓转移中,为胃癌骨髓转移提供新的潜在治疗靶点,同时也发掘出了ACTN2及α-Actinin-2新的医药价值。
附图说明
图1:A为高度侵袭性胃癌患者和非骨髓转移晚期胃癌患者的全身PET-CT结果;B为高度侵袭性胃癌患者和非骨髓转移晚期胃癌患者的组织的转录组分析上调基因结果;C为高度侵袭性胃癌患者与非骨髓转移晚期胃癌患者的基因GO)分析结果;D为差异候选基因在高度侵袭性胃癌患者与非骨髓转移晚期胃癌患者的表达情况;E~F为transwell小室实验和伤口愈合实验检测候选基因对胃癌细胞侵袭迁移能力的影响。
图2:A为胃癌细胞(AGS)中过表达和未过表达ACTN2时,丝状伪足形成情况;B为胃癌细胞中α-Actinin-2表达情况;C为胃癌细胞(AGS)中过表达和未过表达ACTN2时,丝状伪足和板状伪足的分布情况;D为胃癌细胞中降低α-Actinin-2表达时伪足的形成情况。
图3为胃癌细胞(SNU-16)中过表达和未过表达ACTN2时,丝状伪足形成情况。
图4:A为胃癌细胞(AGS)中过表达和未过表达ACTN2时,α-Actinin-2和F-Actin的表达情况;B为胃癌细胞(AGS)中降低α-Actinin-2表达时α-Actinin-2和F-Actin的表达比例;C为α-Actinin 1-4羟基端和羧基端的氨基酸序列;D为胃癌细胞(AGS)中分别过表达α-Actinin 1-4后的表达情况;E为胃癌细胞(AGS)中过表达α-Actinin 2,检测其在细胞膜、细胞质和细胞核中的表达情况。
图5:A为胃癌细胞系和组织中α-Actinin 1、3和4的表达情况;B为胃癌细胞系AGS中敲降α-Actinin 1和4的表达时细胞伪足的数量;C为胃癌细胞系AGS中过表达α-Actinin2时,α-Actinin 1和4的表达量和定位情况;D为胃癌细胞系AGS中过表达α-Actinin 2时,α-Actinin 1、2、4的结合情况;E为α-Actinin 1、4与F-Actin/G-Actin的结合情况;F为α-Actinin 2、4与F-Actin/G-Actin的结合情况。
图6:A为常见转移相关通路中关键转录因子对ACTN2启动子区域活性的影响;B为筛选NF-κB与ACTN2启动子区域结合区域;C为验证NF-κB与ACTN2启动子区域结合位点;D为高度侵袭性胃癌患者和非骨髓转移晚期胃癌患者的肿瘤组织中α-Actinin 2和NF-κB的表达和定位情况;E为α-Actinin 2或/和NF-κB的高表达在高度侵袭性胃癌患者和非骨髓转移晚期胃癌患者的占比及相关性分析;F为α-Actinin 2或/和NF-κB的表达高低对高度侵袭性胃癌患者和非骨髓转移晚期胃癌患者预后的影响;G为p-RelA抗体在IP实验中的特异性和高效性;H为p-RelA与ACTN2基因在细胞核内的结合。
图7:A为敲低α-Actinin 2或RelA对ACTN2转录的影响;B为改变α-Actinin2或RelA表达对NF-κB转录的影响;C为α-Actinin 2入核;D为α-Actinin 2与p-RelA在核内结合;E为降低α-Actinin 2表达时α-Actinin 2与p-RelA在核内结合情况;F为p-RelA与ACTN2基因在核内结合。
图8:A-B为过表达α-Actinin 2的胃癌AGS细胞经心脏注射后在小鼠的骨转移灶;C-D为过表达α-Actinin 2的胃癌AGS细胞经腹腔注射后在小鼠的骨转移灶;E为应用trap染色检测过表达α-Actinin 2的胃癌AGS细胞经心脏注射后形成小鼠的骨转移灶;F为应用trap染色检测过表达α-Actinin 2的胃癌AGS细胞经腹腔注射后在小鼠的骨转移灶;G为经心脏注射形成的骨中trap阳性细胞数;H为经腹腔注射形成的骨中trap阳性细胞数。
具体实施方式
以下通过实施例形式的具体实施方式,对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
实施例1ACTN2与胃癌患者的骨髓转移相关性研究
本实施例筛选了一系列来自伴有弥漫性骨髓转移的高度侵袭性胃癌患者的癌组织(如图1A所示)。随即对高度侵袭性胃癌患者和非骨髓转移的晚期胃癌患者的组织进行了转录组分析。在高度侵袭性胃癌患者中,以阈值FDR□<0.05鉴定了23个基因,这些基因在相邻正常组织和肿瘤样本之间显著过表达。而在晚期胃癌患者的相邻正常组织和肿瘤样本之间以相同的阈值识别出11个显著过表达的基因(如图1B和表1所示)。为了确定与骨转移相关的特定功能过程,进行了GO分析,以可视化高度侵袭性胃癌患者与晚期胃癌患者相比的16个差异表达基因(DEGs)的功能作用(如表1所示)。GO富集分析表明,高度侵袭性胃癌患者上调的DEGs主要富集在角蛋白丝、中间丝细胞骨架和中间丝中(如图1C所示)。在分子功能方面,高度侵袭性胃癌患者上调的DEGs在结构分子活性和细胞动力学结构中显著富集,上述提示细胞骨架和细胞骨架相关蛋白的异常表达是癌细胞导致骨髓转移的关键因素。
表1 Up-regulated genes in HAGC and/or NAGC
进一步通过与《人类蛋白质图谱》的数据进行比较,排除了高度侵袭性胃癌的16个DEGs(https://www.proteinatlas.org/)。ADIPOQ、SPINK6、ACTN2和SYT12在高度侵袭性胃癌中特异性上调且平均FPKM阈值□<1,但在人类蛋白质图谱的数据中未检测到(如图1D和表1所示)。考虑到高度侵袭性胃癌和晚期胃癌之间的明显差异,其中高度侵袭性胃癌细胞具有较强的远处转移能力,将重点放在ADIPOQ、SPINK6、ACTN2、SYT12和FGL1(因为人类蛋白图谱没有显示FGL1的蛋白水平,所以我们将其包括在本实验中)对促进细胞迁移和侵袭的作用上。这五个基因在GC细胞系中过表达(AGS为原位胃癌;SNU-16为腹水胃癌)。在transwell实验和伤口愈合实验中,ACTN2的过度表达增加了细胞运动,而ADIPOQ、FGL1、SPINK6和SYT12对胃癌细胞运动无影响(如图1E和图1F所示)。上述表明ACTN2是胃癌细胞转移的关键调节因子。
实施例2α-Actinin-2通过促进丝状伪足的形成和成熟来促进胃癌细胞迁移
为了评估细胞骨架是否参与α-Actinin-2介导的细胞迁移,本实施例用F-肌动蛋白特异性探针TRITC缀合的磷脂标记细胞骨架的主要成分纤维肌动蛋白(F-Actin)(如图2A所示)。结果表明,内源性α-肌动蛋白-2在粗制小鼠海马中大量表达,但在AGS(人胃腺癌细胞)、SNU-16细胞(人胃癌细胞)中未检测到(如图2B所示)。F-肌动蛋白染色显示α-肌动蛋白-2的过度表达显著增加了几种细胞中丝状伪足的数量和长度,包括AGS(如图2A所示)和SNU-16(如图3所示)。在AGS细胞中,只有13.46%的细胞有一些短丝状伪足(数量:0.199U/细胞;平均长度:1.47μm;)。在α-肌动蛋白-2过度表达的AGS细胞中,超过96%的细胞显示出许多细长的丝状伪足(数量:8.41U/细胞;平均长度:4.69μm;)。α-肌动蛋白-2的过度表达使该蛋白在全细胞分布,α-肌动蛋白-2分布在细胞核、细胞质、膜和丝状伪足中(如图2C所示)。然而,α-肌动蛋白-2定位于丝状足近端,并从近端到远端逐渐减少(如图2C所示)。这些结果表明,α-肌动蛋白-2是丝状伪足形成所必需的。
在α-Actinin-2过表达2天后,用siRNA敲低α-Actinin-2抑制了丝状伪足的增加。对三个随机选择的细胞组的分析显示,在α-Actinin-2敲低后,丝状伪足的数量和长度显著减少(如图2D所示)。这表明α-Actinin-2是丝状伪足成熟所必需的。
使用激光共聚焦检测,观察到除细胞膜和丝状伪足近端区域外,在细胞的任何部分均未观察到过表达的α-Actinin-2和磷脂标记的F-肌动蛋白的显著共定位(如图4A)。F-肌动蛋白在GC细胞的细胞膜上均匀分布,在α-肌动蛋白-2过度表达时,在丝状足的近端部分局部聚集(如图4A所示)。F-肌动蛋白复合物是由球状肌动蛋白单体(G-Actin)组装而成的聚合物。为了确认α-Actinin-2是否通过G-肌动蛋白组装促进丝状伪足的形成和成熟,测量了这种蛋白的两种形式的比率。发现α-Actinin-2的过度表达增加了F-肌动蛋白与G-肌动素的比率,而α-Actinin-2的敲除降低了这一比率(如图4B所示)。上述表明,具有骨向性的胃癌细胞在F-肌动蛋白增强的情况下更为坚硬。进一步采用免疫共沉淀法分析α-Actinin-2与肌动蛋白之间的相互作用。使用了对α-Actinin 1、2、3或4具有特异性的抗体(这些抗体的免疫原如图4C和4D所示),并且在通过过度表达GFP-α-Actinin1、2、3或4。使用特异性抗体,免疫共沉淀分析显示,α-Actinin-2和肌动蛋白主要在细胞的质膜成分中形成稳定的复合物,但在细胞质和细胞核中仅形成少量α-肌蛋白-2/肌动蛋白复合物(如图4E所示)。这些数据表明,α-肌动蛋白-2直接结合肌动蛋白并促进其组装成丝状伪足。
实施例3α-Actinin-2通过替换α-Actinin-1形成α-Actinin-2:α-Actinin-4复合物来增强F-Actin结合能力
在人类中发现了α-肌动蛋白的多种同工型,并由至少四个不同的基因编码。常见的GC细胞含有两种非肌肉α-Actinin同种型,α-Actinin-1和α-Actinin-4(如图5A所示)。但是,使用免疫印迹未检测到骨骼肌特异性的α-肌动蛋白同工型α-Actinin-2和α-Actinin-3(如图5A所示)。而在α-Actinin-2过表达2天后,用特异性siRNA敲低α-Actinin-4而不是α-Actinin-1抑制了丝状伪足的数量和长度的增加(如图5B所示),表明α-Actinin-4是α-Actinin-2诱导的丝状伪足形成所必需的。
实验发现,α-Actinin-2过表达后α-Actinin-1和α-Actinin-4/F-Actin之间的共定位降低(如图5C所示);α-Actinin-2的过表达诱导了α-Actinin-2,α-Actinin-4和F-Actin的显著共定位(如图4A和图5C所示)。同时,α-Actinin-1和α-Actinin-2之间的共定位也发生在细胞质膜上(如图5C所示)。进一步测试了α-肌动蛋白之间的直接相互作用。在α-Actinin-1下拉后检测到α-Actinin-4和少量肌动蛋白,反之亦然(如图5D所示)。在α-Actinin-2过表达条件下,α-Actinin-1下拉后检测到α-Actinin-2,但肌动蛋白和α-Actinin-4低于检测限;在α-Actinin-2下拉后,检测到广泛的α-Actinin-4,Actin和少量α-Actinin-1;在α-Actinin-4下拉后检测到丰富的α-Actinin-2和Actin,但未检测到α-Actinin-1。因此,α-Actinin-2的上调导致α-Actinin-二聚体组成从α-Actinin-1:α-Actinin-4转变为α-Actinin-2:α-Actinin-4和α-Actinin-2:α-Actinin-1。
进一步使用通常用于鉴定肌动蛋白相关蛋白的肌动蛋白共沉淀测定法评估了α-Actinin-2:α-Actinin-4复合物是否发挥了与F-Actin结合的基本作用。在α-Actinin-2:α-Actinin-4复合物中检测到强F-肌动蛋白结合,而在α-Actinin-1:α-Actinin-4复合物中检测到低F-Actin结合活性(如图5E所示)。此外,在α-Actinin-1:α-Actinin-4和α-Actinin-2:α-Actinin-4复合物中均未检测到G-Actin的广泛结合(如图5E所示)。结果表明,α-Actinin-2通过取代α-Actinin-1形成α-Actinin-2:α-Actinin-4复合物来增强F-Actin结合能力。
实施例4ACTN2是GC细胞NF-κB信号传导的直接靶基因
研究发现,AKT/GSK3β/c-Fos/NFATc1,IGF-1/AKT/NF-κB(RelA)或TGF-β/Smad3/4信号级联在许多癌症的BMM中起关键调节作用。为了证明哪种转录因子可以调节ACTN2基因启动子的激活,本实施例将c-Fos,NFATc1,RelA,Smad3或Smad4与ACTN2启动子(相对于转录起始位点的-1918至+2455的核苷酸)报告基因共转染到胃癌细胞中。结果显示,只有RelA,显著激活ACTN2的启动子活性(如图6A所示),证明了NF-κB依赖性调节α-Actinin-2表达的方式。
本实施例进一步鉴定了启动子活化所需的核心序列。将包含ACTN2启动子5'侧翼区域缺失的一系列报道基因构建体克隆到报道基因载体中。所有截短的ACTN2启动子构建体均具有相同的3'端。其中一个缺失构建体(核苷酸+331至+2455)显示出与原始构建体(核苷酸-1918至+2455)相同的响应,而另一个缺失构建体(核苷酸+616至+2455)显示几乎完全消除了RelA诱导的启动子活动(如图6B所示)。因此,主要的ACTN2转录控制元件被缩小到跨越核苷酸+331至+616的区域。启动子分析鉴定了人ACTN2启动子核心区域中潜在的RelA结合位点+541/+550(TGC AAT TTC C),并且该位点的突变显示响应于RelA处理的ACTN2启动子活化的明显破坏(如图6C所示)。同时,进行免疫染色以检测胃癌组织中α-Actinin-2和Ser276-磷酸化RelA(p-RelA)的细胞共定位。与常规晚期胃癌组相比,高度侵袭性胃癌组显示出显著更高的α-Actinin-2和p-RelA表达,并且α-Actinin-2阳性细胞与p-RelA高表达细胞重叠,其中活化的p-RelA亚基定位于细胞核和α-Actinin-2蛋白主要位于同一细胞的膜中(如图6D和图6E所示)。详细的患者特征和临床病理参数显示在表2中。基于单变量分析,相关性分析显示任何α-Actinin-2和p-RelA的高表达与较差的存活相关(如图6E所示)。α-Actinin-2和p-RelA的同时高染色与高度侵袭性胃癌中的总体存活显着相关,但与常规晚期胃癌中的总体存活无关。具有α-Actinin-2和p-RelA高表达的高度侵袭性胃癌患者的预后最差(如图6F所示)。这些结果表明p-RelA和α-Actinin-2之间存在密切关联。
为了证实体内RelA和ACTN2启动子之间的直接相互作用,在存在或不存在RelA过表达的情况下,在胃癌细胞中使用ChIP测定。染色质用p-RelA抗体沉淀。在RelA过表达细胞中容易检测到与DNA交联的p-RelA蛋白(如图G所示)。通过PCR用跨越RelA结合位点+541/+550(引物:Fs/Rs)或ACTN2启动子上远离该位点的引物(引物:Ff/Rf)的引物通过PCR测定沉淀的DNA。来自RelA过表达细胞的DNA片段通过PCR引物Fs/Rs强烈扩增,但不通过PCR引物Ff/Rf强烈扩增(如图H所示)。上述表明,ACTN2是GC细胞中RelA/NF-κB信号传导的直接靶基因。
表2
Clinical characteristics of patients detected by multipleimmunofluorescence
staining ofα-Actinin-2and p-RelA
Chi-square test or Fisher’s exact test were used in Table,boldindicates P<0.05.ECOG PS,Eastern Cooperative Oncology Group PerformanceStatus.*Grading according to WHO:G1(Well differentiated),G2(Moderatelydifferentiated),G3(Poorly differentiated or undifferentiated).
实施例5NF-κB激活α-Actinin-2正向调节环路增强ACTN2基因转录
实验发现,全长α-Actinin-2的过表达仅引起ACTN2启动子或RelA反应报告基因的弱激活(如图7A和图7B所示)。然而,与单独的RelA相比,α-Actinin-2与RelA的共同表达增强,而α-Actinin-2的敲除显著降低了荧光素酶活性,并且RelA敲除几乎可以完全阻断α-Actinin-2和RelA诱导的荧光素活性。这些数据表明,α-Actinin-2特异性共激活RelA,并可能潜在地促进其自身转录的阳性自动调节环。
α-Actinin-2作为转录辅激活物的功能必须与核分布相关。在高度侵袭性组织细胞和α-Actinin-2过表达细胞中观察到α-Actinin-2的区域性或点状核定位(如图7C和图2C所示)。通过从细胞中提取核蛋白,我们确认了α-Actinin-2在细胞核中定位并与p-RelA形成复合物(如图7D所示)。
为了确定α-Actinin-2通过ACTN2基因的自动调节是直接的还是间接的,使用ChIP分析研究了存在或不存在RelA过度表达、α-Actinin-2过度表达和/或α-Actinin-2特异性siRNA的胃癌细胞。用RelA抗体沉淀染色质。α-Actinin-2和与DNA交联的RelA蛋白很容易在沉淀物中检测到(如图7E所示)。我们发现RelA结合位点周围的DNA片段可以从RelA过表达细胞中扩增出来。α-Actinin-2的共表达增强,但α-Actinin-2特异性siRNA降低了这种扩增(如图7F所示),证实NF-κB中α-Actinin-2通过正向自动调节环路触发ACTN2基因转录。
实施例6α-Actinin-2在体内促进原发性肿瘤生长和骨髓转移
本实施例将组成型表达α-Actinin-2的生物发光AGS细胞(1×106个细胞)注射到无胸腺裸鼠的左心室中。左心室的心内注射导致肿瘤细胞最初分布在整个动脉系统中并测试肿瘤细胞定殖所有器官的能力。每周用IVIS成像系统定期监测小鼠。与对照相比,α-Actinin-2在实验过程中通过生物发光成像确定显著增加了原发肿瘤的大小(如图8A所示)。鉴于α-Actinin-2对体外胃癌细胞迁移的强大扩展作用,认为它也可能促进体内转移。心内注射后2周,α-Actinin-2诱导骨自发转移,而对照细胞未观察到转移(如图8A和图8B所示)。α-Actinin-2过表达细胞生长积极,在骨盆和股骨中观察到转移性病变,但在其他器官中没有观察到转移性病变,表明α-Actinin-2的高效率和特异性,诱导胃癌细胞的骨髓转移。为了证实α-Actinin-2促进骨髓转移的特性,将组成型表达α-Actinin-2的细胞腹膜内注射到无胸腺裸鼠中(如图8C所示)。实际上,α-Actinin-2仅在腹膜内注射后11天诱导骨转移,而对照细胞未观察到转移(如图8C和图8D所示)。此外,在骨盆和股骨中也观察到α-Actinin-2过表达细胞,但在其他器官中未观察到。
采用破骨细胞的TRAP染色鉴定α-Actinin-2驱动的骨髓转移。与对照组相比,心内注射小鼠和腹膜内注射小鼠的α-Actinin-2组中的TRAP阳性细胞增加(如图8E-H所示)。α-Actinin-2心内和腹腔注射组的破骨细胞分布相似,但心内注射显著增加破骨细胞(如图8G和图8H所示)。这些结果不仅表明α-Actinin-2在促进骨髓转移方面非常有效,而且还表明α-Actinin-2可以诱导骨髓转移而不依赖于血行转移。
最后应当说明的是,以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制,尽管参照较佳实施例对本发明作了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的实质和范围。
Claims (9)
1.检测ACTN2的试剂在制备预测或检测胃癌骨髓转移的产品中的应用;所述应用为非疾病诊断和治疗的应用。
2.根据权利要求1所述的应用,其特征在于,所述产品包括检测样本中ACTN2的表达量的试剂。
3.根据权利要求2所述的应用,其特征在于,所述样本中ACTN2的表达量上调。
4.根据权利要求2所述的应用,其特征在于,所述试剂包括通过PCR或免疫检测检测ACTN2表达水平的试剂。
5.根据权利要求1所述的应用,其特征在于,所述产品包括芯片或试剂盒。
6.检测ACTN2编码的蛋白的试剂在制备预测或检测胃癌骨髓转移的产品中的应用。
7.根据权利要求6所述的应用,其特征在于,所述蛋白为α-Actinin-2。
8.根据权利要求7所述的应用,其特征在于,所述产品包括检测样本中α-Actinin-2的表达量的试剂。
9.根据权利要求8所述的应用,其特征在于,所述样本中α-Actinin-2的表达量上调。
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