CN116718784A - Stc1作为胶质瘤标记物的应用 - Google Patents
Stc1作为胶质瘤标记物的应用 Download PDFInfo
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Abstract
本发明提出了STC1作为胶质瘤标记物的应用;所述胶质瘤标记物STC1可以作为胶质瘤生物标记物和治疗靶点,为制备胶质瘤辅助诊断、疗效检测、预后判断相关药物,以及为制备抑制胶质瘤增殖药物提供新的方向;同时还可以用于抑制胶质瘤药物的筛选提供更多的选择。
Description
技术领域
本发明属于胶质瘤标记物技术领域,具体涉及STC1作为胶质瘤标记物的应用。
背景技术
胶质瘤是常见的颅内肿瘤之一,其发病率占颅内原发性肿瘤的40%~60%,具有高侵袭性、高复发率、预后差等特点。胶质母细胞瘤(glioblastoma,GBM)是胶质瘤中恶性度最高的,其中位生存期仅有12个月,5年生存率不足5%;目前临床上对胶质瘤的治疗以综合治疗为主,手术切除后通常辅以放、化疗。替莫唑胺(Temozolomide;TMZ)作为第二代口服烷基化剂的抗肿瘤药物,可透过血脑屏障,主要用于治疗成人顽固性胶质瘤及复发或进展性的多形胶质母细胞瘤,其抗肿瘤的主要机制为烷基化导致DNA损伤及错配修复失败产生的细胞毒作用。内源性O6-甲基鸟嘌呤-DNA甲基转移酶(O6-methylguanine DNAmethyltransferase,MGMT)能与DNA鸟嘌呤第6位氧原子上的烷基化合物结合,将烷基转移到MGMT的第145号半胱胺酸活性位上,使DNA上烷基化的鸟嘌呤被还原,从而降低替莫唑胺对肿瘤细胞的杀伤效果,是替莫唑胺常见的耐药机制之一。
斯钙素-1(Stanniocalcin-1,STC1)是一种最早在鱼类中发现的糖激素蛋白,主要参与钙和磷酸盐平衡的调节。在随后的研究中发现,STC1(NCBI登录号NM_003155.3,GeneID:6781)在哺乳动物的多种组织中表达,并分泌至胞外后发挥多种功能,如抑制钙离子转运、刺激磷酸盐重吸收、抗凋亡、抗炎、致癌、促进血管生成、参与葡萄糖代谢等。而且,STC1在肿瘤组织中也会高表达,越来越多的研究表明STC1似乎在肿瘤进展中起着重要作用。STC1高表达可以通过JNK途径增加肝癌细胞和乳腺癌细胞的侵袭能力,促进转移的发生。
研究显示,STC1在胶质瘤发生发展中发挥着重要的作用。STC1可以通过自分泌途径,与NOTCH受体胞外端的EGF样重复序列结合并激活NOTCH1来促进GBM细胞的干细胞特征。研究表明,降低STC1的表达可以抑制GBM细胞的增殖和侵袭能力,而过表达STC1则会增加GBM细胞的增殖和侵袭能力,并且STC1作为microRNA调控的靶基因还参与了GBM细胞的转移调控。
发明内容
申请人在研究中发现胶质瘤患者中STC1异常高表达,且随着胶质瘤级别增加表达更显著,STC1高表达与胶质瘤患者不良预后显著相关。通过构建STC1慢病毒载体建立STC1敲降的细胞模型,研究其对胶质瘤细胞中生长和替莫唑胺疗效的影响。旨在探讨STC1的表达与胶质瘤发生发展、化疗耐药的关系,为进一步研究STC1对胶质瘤的靶向治疗及调控机制奠定基础。并且用shRNA敲降STC1表达对胶质瘤显著抑制,提示STC1可以作为胶质瘤生物标记物和治疗靶点,并用于胶质瘤辅助诊断、疗效检测、预后判断提供新的方向。
有鉴于此:
本发明的第一个目的在于,提供胶质瘤标记物STC1在制备胶质瘤辅助诊断、疗效预测及预后判断试剂中的应用。
所述的,胶质瘤辅助诊断包括但不限于胶质瘤的分级诊断、侵袭或转移诊断等;疗效预测包括但不限于胶质瘤减轻或加重、治愈或恶化、预判胶质瘤转移风险、诊断胶质瘤是否发生转移等预测;预后判断包括但不限于胶质瘤是否复发、判断胶质瘤转移是否复发。具体通过检测组织中STC1的表达水平来实现诊断、预测或判断。
进一步地,所述试剂包括特异性结合STC1蛋白的抗体或蛋白质芯片,特异性检测STC1 mRNA的引物、探针、核酸芯片。
进一步地,所述胶质瘤为新鲜的、冷冻的或石蜡固定包埋的组织。
本发明的第二个目的在于,提供STC1表达抑制剂在制备抑制胶质瘤细胞增殖药物中的应用。
进一步地,所述抑制剂选自以STC1基因或其转录本为靶序列、且能够抑制其蛋白表达或基因转录的反义核酸;或能表达或形成所述反义核酸的构建物。
优选地,所述抑制剂具备STC1静默或敲低功能,包括miRNA、siRNA、dsRNA或shRNA;或包含所述miRNA、siRNA、dsRNA或shRNA的过表达质粒载体或慢病毒。
进一步地,所述抑制胶质瘤细胞增殖药物包括胶质瘤化疗药物和/或药学上可接受的载体。优选地,所述胶质瘤化疗药物包括替莫唑胺;替莫唑胺和STC1抑制剂具有协同抑制胶质瘤细胞增殖的效果。
本发明的第三个目的在于,提供胶质瘤标记物STC1在体外筛选治疗和/或预防胶质瘤药物、预防胶质瘤转移、侵袭药物中的应用。
进一步地,所述治疗和/或预防胶质瘤药物、预防胶质瘤转移、预防胶质瘤侵袭药物能够抑制STC1高表达。
在本发明的上下文中,“STC1基因”包括STC1基因以及STC1基因的任何功能等同物的多核苷酸。STC1基因包括与NCBI登录号NM_003155.3,Gene ID:6781的DNA序列具有70%以上同源性,且编码相同功能蛋白质的DNA序列;
优选地,STC1基因的编码序列包括以下任意一种DNA分子:
(1)序列表中SEQ ID NO:1所示的DNA序列;
(2)在严格条件下与(1)限定的DNA序列杂交且编码相同功能蛋白质的DNA序列;
与(1)或(2)限定的DNA序列具有70%、优选地,90%以上同源性,且编码相同功能蛋白质的DNA分子。
在本发明的具体实施方案中,所述STC1基因的编码序列是SEQ ID NO:1所示的DNA序列。
在本发明的上下文中,STC1基因表达产物包括STC1蛋白以及STC1蛋白的部分肽。
“STC1蛋白”包括STC1蛋白以及STC1蛋白的任何功能等同物。所述功能等同物包括STC1蛋白保守性变异蛋白质、或其活性片段,或其活性衍生物或其突变体。突变体包括等位变异体、天然突变体、诱导突变体、其氨基酸序列通过缺失、替代、增加和/或插入发生变异的突变体、与修饰的氨基酸序列功能相同的突变体及在高或低的严格条件下能与STC1的DNA杂交的DNA所编码的蛋白质。
相比现有技术,本发明的有益效果在于:
1.本发明提供的标记物STC1可以作为胶质瘤生物标记物和治疗靶点,为制备胶质瘤辅助诊断、疗效检测、预后判断相关药物提供新的方向。
2.本发明提供的标记物STC1为制备抑制胶质瘤增殖药物提供新的方向;同时还可以用于抑制胶质瘤药物的筛选,为胶质瘤临床治疗应用提供更多的选择。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1为CGGA数据库中原发胶质瘤患者STC1高表达和低表达患者的生存曲线的比较(A),CGGA数据库中复发胶质瘤患者STC1高表达和低表达患者的生存曲线的比较(B),CGGA数据库中替莫唑胺化疗患者和未经替莫唑胺化疗患者STC1表达量比较(C)。
图2为RT-qPCR检测STC1在胶质瘤临床样品与癌旁组织临床样品的表达量比较(A),CGGA数据库中不同级别胶质瘤患者STC1表达量比较(B),免疫荧光所示STC1在不同级别胶质瘤中的表达量差异(C)。
图3为RT-qPCR验证STC1过表达细胞模型(A),蛋白免疫印迹实验验证STC1过表达细胞模型(B),RT-qPCR验证STC1敲降细胞模型(C),蛋白免疫印迹实验验证STC1敲降细胞模型(D)。
图4为克隆形成实验检测替莫唑胺处理STC1过表达/野生型细胞克隆形成率的比较(A),克隆形成实验检测替莫唑胺处理STC1敲降/野生型细胞克隆形成率的比较(B)。
图5为彗星电泳实验检测替莫唑胺处理STC1过表达/野生型细胞后DNA损伤水平的比较(D),彗星电泳实验检测替莫唑胺处理STC1敲降/野生型细胞后DNA损伤水平的比较(D)。
图6为流式细胞术检测替莫唑胺处理STC1过表达/野生型细胞后细胞凋亡比率的比较(A),流式细胞术检测替莫唑胺处理STC1敲降/野生型细胞后细胞凋亡比率的比较(B)。
图7为蛋白免疫印迹实验检测替莫唑胺处理STC1过表达/野生型细胞后细胞凋亡、DNA损伤相关蛋白的表达量比较(A-B),蛋白免疫印迹实验检测替莫唑胺处理STC1敲降/野生型细胞后细胞凋亡、DNA损伤相关蛋白的表达量比较(C-D)。
图8为化学发光法检测STC1过表达/野生型细胞在裸鼠颅内原位建模后经60mk/Kg替莫唑胺处理不同时间点肿瘤增殖的代表性图片(A)及肿瘤细胞增殖速率的比较(B),化学发光法检测STC1敲降/野生型细胞在裸鼠颅内原位建模后经60mk/Kg替莫唑胺处理不同时间点肿瘤增殖的代表性图片(C)及肿瘤细胞增殖速率的比较(D)。
图9为HE染色比较STC1过表达/野生型细胞在裸鼠颅内原位建模后经60mk/Kg替莫唑胺治疗的肿瘤体积(A),HE染色比较STC1敲降/野生型细胞在裸鼠颅内原位建模后经60mk/Kg替莫唑胺治疗的肿瘤体积(B)。
具体实施方式
下面将结合本发明实施例对本发明的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
若未特别指明,实施例中所用的实验手段为本领域技术人员所熟知的常规手段。
实施例1:通过CGGA数据库的数据分析,挖掘STC1基因在胶质瘤患者中的表达差异与生存期的关系,STC1表达与胶质瘤患者化疗状态的关系
采用CGGA标准方法下载胶质瘤患者组织样本的RNA-seq测序数据和临床资料,根据原发性和复发性将胶质瘤患者分为两类,每一类根据STC1表达水平的高低对患者的生存期进行统计分析;另一方面,根据患者是否接受替莫唑胺化疗将其分为两组,统计分析各组患者组织中STC1的表达差异。
结果:见图1A和1B分析发现与STC1低表达患者相比较,STC1高表达患者生存率显著降低,考虑可作为胶质瘤的预后判断指标,图1C显示STC1在替莫唑胺化疗的胶质瘤患者中表达量高于未经替莫唑胺化疗的患者。
实施例2:RT-qPCR和免疫荧光验证STC1基因在胶质瘤组织和非肿瘤组织中的表达差异
通过测定三对胶质瘤组织和癌旁组织中的STC1基因表达状态,胶质瘤组织通过外科手术切除并已获得病理诊断证实。
首先,进行组织细胞总RNA提取,所有操作及相关试剂置于冰浴中操作。使用Trizol试剂提取细胞总RNA,步骤如下:在组织中加入适量Trizol试剂,置于冰上用高速电动研磨仪将组织破碎,充分裂解后,使用高速冷冻离心机4℃12000rpm离心5min,弃沉淀。按200ul氯仿/ml Trizol加入氯仿,振荡混匀15min,室温放置15min。再次使用高速冷冻离心机4℃12000g离心15min。然后吸取上层水相,至另一离心管中。按0.5ml异丙醇Trizol加入异丙醇混匀,室温放置10min。再次使用高速冷冻离心机4℃12000g离心10min,弃上清,RNA沉于管底。加入1ml 75%乙醇,温和振荡离心管,悬浮沉淀。使用高速冷冻离心机4℃8000g离心5min,尽量弃上清,将残留的RNA在超净工作台下晾干3min,并加入30ul H2O溶解RNA样品。通过测OD值测定RNA的质量和浓度,A260/A280值在1.6-1.8之能够确保RNA的纯度。
接着用HiScript II Q RT SuperMix for qPCR试剂盒(诺维赞)合成cDNA。具体步骤如下:按照试剂盒说明书按浓度加入PCR管中最后加入RNA样品。在PCR仪中设置反应条件为:50℃15min(反转录),85℃15s(热激终止反应)得到cDNA样品。
然后用ChamQ SYBR qPCR Master Mix试剂盒(诺维赞)鉴定STC1基因的表达量。具体操作步骤如下:按照表1所示试剂及浓度加入PCR反应管中最后加入cDNA样品。在PCR仪中设置反应条件为:95℃30s(变性),95℃10s(退火),60℃30s(延申),共40个循环,以GAPDH作为相对定量内参,PCR引物见表2.
表1.RT-PCR鉴定STC1基因表达量的方法
表2.RT-PCR鉴定STC1基因的引物序列
数据处理与分析:用2-ΔΔCt法计算基因相对表达量。
结果:如图2A,我们检测了STC1在胶质瘤组织和癌旁组织中的mRNA表达量,发现STC1 mRNA在肿瘤组织中高表达,验证了数据库的结论。
实施例3:通过免疫荧光染色,挖掘STC1表达与胶质瘤患者临床分级的关系。
将临床收集到的胶质瘤样本置于10%的中性缓冲福尔马林,一般固定液的量为组织块体积的4~10倍,在常温条件下固定8~24h后放入30%的蔗糖溶液中脱水24h,用冰冻切片包埋机包埋后切片,载玻片选择粘附载玻片,厚度5μm,破膜0.1%Triton x-100浸泡10min,PBS浸泡5min×2次,取出切片,擦干组织周围液体后滴加10%BSA封闭液,37℃孵育40分钟。滴加一抗(浓度配比,PBS稀释1:150,20ul,保持组织湿润状态但表面不能有液体,放入专用孵育盒内,4℃冰箱过夜后取出切片,PBS缓冲液中清洗3分钟×3次,擦干组织周围液体后滴加荧光二抗,37℃温箱内孵育40分钟;PBS缓冲液中清洗3分钟×3次,擦干组织周围液体后滴加含有DAPI的抗荧光淬灭封片剂。
结果:如图2C免疫荧光染色结果显示STC1在胶质瘤中表达显著高于癌旁组织,并且随分级增加表达量也增加;与CGGA数据库中的结论一致(图2B)。
实施例4:利用慢病毒过表达和敲降STC1基因,探究其表达水平对胶质瘤细胞增殖及化疗的影响。
STC1过表达/敲降GBM细胞模型构建及验证;将shSTC1靶向序列(见表3)和STC1的表达片段(见上述STC1序列信息)经过酶切、连接等步骤分别克隆至21915载体(Dox诱导表达型载体)和pLvx-puro载体;获得重组质粒后,通过目的质粒与慢病毒助包装质粒共转染HEK-293T细胞获得慢病毒:
用500μl opti-MEM稀释三种质粒,共10μg:
PsPAX2 3.3μg
pcmv-VSVG 2.2μg
核心质粒4.4μg
用500mL opti-MEM稀释30μL PEI(1mg/ml);
将质粒混合液加入稀释后的PEI中,轻柔地混匀,静置5min;将质粒、转染试剂混合液均匀地滴加在细胞密度为80%的HEK-293T细胞培养液中;6h后HEK-293T细胞更换10mL完全培养液,培养箱中培养(37℃、5%二氧化碳),48h后收集HEK-293T细胞的上清液至15ml离心管中,2000r/min离心10min,用0.22μm滤器过滤后取1mL病毒液加入目标细胞培养液中,同时加入1μl聚凝胺(polyberen,1mg/mL),混合均匀后培养箱中培养(37℃、5%二氧化碳),48h后观察转染效率,后续进行流式细胞仪分选或嘌呤霉素筛选,获得STC1过表达/敲降GBM细胞模型。
表3.慢病毒载体表达shRNA敲降STC1基因的引物序列
结果:如图3A-B显示,利用慢病毒过表达载体构建的STC1过表达细胞模型中,STC1基因在mRNA水平和蛋白质水平均显著上调;利用shRNA干扰技术敲降STC1的细胞模型中,STC1基因在mRNA水平和蛋白质水平均显著下降,如图3C-D。
实施例5:利用克隆形成实验比较STC1表达水平对细胞增殖和替莫唑胺化疗的影响。
400个细胞/孔,铺12孔板,每组细胞各3个重复孔;细胞贴壁后,根据实验设计每组细胞分别给予安慰剂/TMZ处理,每隔72h更换一次药物,10-14天后对细胞用10%中性福尔马林固定10min、PBS洗一遍,加入4%结晶紫染色液室温染色15min后去离子水清洗一遍,晾干后拍照统计分析各组细胞克隆形成率,评估STC1表达水平对TMZ抑制细胞生长的影响。
结果:如图4A所示,过表达STC1,促进胶质瘤细胞的生长,而且表现出对替莫唑胺化疗的耐受;而敲降STC1会抑制胶质瘤细胞的增殖,同时会提高替莫唑胺对胶质瘤细胞的抑制作用,如图4B。
实施例6:利用彗星电泳实验比较STC1表达水平对替莫唑胺化疗的影响。
将各组经安慰剂和TMZ处理后细胞消化计数,制备成2*10^4个/mL单细胞悬液;取45℃水浴保温的0.5%正常熔点琼脂糖100μL铺于载玻片上,盖玻片推匀,放入4℃凝固后形成底胶;水平取下盖片,取37℃水浴中保温的0.5%低熔点琼脂糖100μL与20μL细胞悬液混匀,立即加上盖玻片,4℃凝固5至8min;将制备好的胶板去掉盖玻片后,浸于4℃预冷的细胞裂解液中裂解2-3h;然后浸泡在4℃预冷的电泳液中解旋20min;载玻片水平放置在电泳槽阳极端附近,4℃电泳20-25min(25V,300mA);将胶板进行中和置于染色缸中,在2μg/ml的EB染色液中,暗处染色5-10min后拍照观察,统计DNA损伤细胞的比例,评估STC1的表达水平对TMZ化疗导致细胞DNA损伤的影响。
结果:如图5A显示,STC1高表达对细胞DNA没有影响,经过相同浓度的替莫唑胺处理相同时间,野生型细胞表现出较高水平的DNA损伤,STC1过表达的细胞中DNA损伤比率相对较低,而且这种差异具有同统计学意义;另一方面,敲降STC1对DNA没有影响,经过相同浓度的替莫唑胺处理相同时间,野生型细胞表现出较高水平的DNA损伤,STC1敲降的细胞中DNA损伤比率相对更高,表明:敲降STC1可以提高替莫唑胺的化疗疗效(如图5B)。
实施例7:利用流式细胞仪分析STC1表达水平对替莫唑胺诱导的细胞凋亡比例的差异。
将各组细胞消化计数后铺在12孔板中,1*10^5个/孔,分别给予安慰剂和TMZ处理72小时,然后根据凋亡染色分析试剂盒说明书进行凋亡细胞染色,通过流式细胞仪分析各组细胞经安慰剂和TMZ处理后的凋亡比例评估GBM细胞过表达/敲降STC1对TMZ化疗诱导细胞凋亡的影响。
结果:如图6A所示,经过相同浓度的替莫唑胺处理相同时间,野生型细胞模型中细胞凋亡比例显著升高,而STC1高表达细胞模型中细胞凋亡比例升高不显著;当敲降STC1基因表达后,替莫唑胺诱导的细胞凋亡水平是野生型细胞的1.5倍(图6B)。以上结果表明,过表达STC1导致胶质瘤细胞对替莫唑胺耐受,敲降STC1可以显著提升替莫唑胺的化疗疗效。
实施例8:利用蛋白免疫印迹实验比较STC1表达水平对细胞增殖、化疗耐受相关蛋白表达的差异。
按照蛋白提取试剂盒说明书提取细胞样品中的总蛋白,定量检测各组细胞蛋白浓度,变性处理后10μg/孔上样;按照80V,30min,110V,60min电泳对目的蛋白进行分离,然后20V,60min将蛋白通过半干式转印仪转印至PVDF膜上,5%的脱脂奶粉封闭2h后,4℃过夜孵育第一抗体,洗涤PVDF膜后,常温孵育对应种属的第二抗体,再次洗涤PVDF膜,然后通过化学发光检测各组细胞中目的蛋白的表达,以GAPDH为内参计算目的蛋白的相对表达量。
结果:如图7A-B所示,在野生型/高表达STC1细胞模型中给予替莫唑胺治疗,凋亡蛋白Caspase-3和PARP1在野生型细胞表达显著上调,DNA损伤修复蛋白γ-H2AX表达增加,而STC1高表达细胞中无显著变化;如图7C-D所示,野生型/敲降STC1的细胞给予替莫唑胺治疗后,凋亡蛋白Caspase-3和PARP1、DNA损伤修复蛋白γ-H2AX在野生型细胞表达均上调,但在STC1敲降的细胞模型中升高更加显著。
实施例9:应用STC1过表达/敲降细胞构建的裸鼠颅内原位成瘤模型,研究STC1的表达水平对TMZ疗效的影响。
①颅内原位成瘤建模:取生长良好的STC1过表达/敲降的GBM细胞,弃去原培养基,PBS漂洗2次。用胰蛋白酶(0.25%,含EDTA)约1mL消化细胞,DMEM完全培养基终止消化后离心去除上清,加入PBS重悬洗涤细胞,计数后离心,再加入适量PBS重悬以调整细胞浓度,放于冰盒中备用。1%戊巴比妥钠按50mg/kg给裸鼠进行腹腔注射,待裸鼠麻醉后,通过耳杆和上齿固定器固定于小鼠脑立体定位仪上,消毒后切开头顶皮肤,暴露前囟,取其后方3mm,右侧距中线2mm处钻孔。使用10μL微量进样器吸取4μL细胞悬液(含5×104个细胞)固定于立体定向仪且对准钻孔处,缓慢垂直进针3mm,退针0.5mm,缓慢注入细胞悬液,每注射1μL暂停1min。注射完毕后暂停3min再缓慢拔针,以骨蜡封闭骨窗,逐层缝合关颅。
②TMZ治疗及动物模型检测:模型建立后第10天,过表达/敲降STC1的GBM细胞构建的动物模型中(n=8),分别给予安慰剂和60mg/kg的TMZ灌胃治疗,每周给药5天,休息两天,连续治疗4周;对照组给予相同剂量的生理盐水灌胃,治疗周期同实验组。检测颅内原位成瘤模型时,提前30min打开小动物活体成像仪,然后给各组裸鼠称量体重,根据裸鼠体重腹腔注射荧光素(Luciferase的底物,按每克注射10μL 15mg/mL的荧光素),等待8-10min使酶和底物充分反应后将裸鼠置于通有异氟烷气体的麻醉箱内麻醉,有必要可调整空气与异氟烷气体的流入量。裸鼠被完全麻醉后,关闭通入麻醉箱的麻醉气体,打开成像仪暗箱的麻醉气体通道,将裸鼠放置于暗箱平台中(裸鼠头应嵌入异氟烷导流槽内,防止其中途醒来影响成像结果),在腹腔注射荧光素后的第12min检测裸鼠体内发出的生物发光,最后用软件获取和整理图像,记录ROI值。各组动物模型在治疗的同时,每隔4天称量体重、通过小动物活体成像检测颅内化学发光值,绘制各组裸鼠的生存曲线。
结果::STC1高表达细胞原位移植瘤荧光强度大小见图8A,如图8B,STC1高表达的动物模型中ROI值统计曲线增值速率显著高于对照组动物模型;而同样给予60mg/kg的替莫唑胺治疗,STC1过表达组的肿瘤增殖曲线仍然高于对照组动物模型,表明STC1高表达促进胶质瘤的增殖和替莫唑胺耐受。STC1敲降细胞原位移植瘤荧光强度大小见图8C,如图8D,STC1敲降细胞的动物模型中ROI值统计曲线增值速率显著低于对照组动物模型;给予60mg/Kg的替莫唑胺治疗,STC1敲降细胞的肿瘤增殖曲线显著低于对照组动物模型,表明敲降STC1表达,可以抑制胶质瘤细胞增殖、增强替莫唑胺化疗疗效。
实施例10:通过HE染色比较各组颅内原位动物模型中肿瘤体积的差异。
将各组动物实验获取的脑组织置于10%的中性缓冲福尔马林,在常温条件下固定24h后脱水,切片,载玻片选择粘附载玻片,厚度2μm,烤片温度设置65~68℃,时间为2h,依次将切片放入:二甲苯I10min→二甲苯II 10min→无水乙醇Ⅰ5min→无水乙醇Ⅱ5min→95%酒精5min→90%酒精5min→80%酒精5min→70%酒精5min→蒸馏水洗;然后切片入Harris苏木素染3~8min,自来水洗,1%的盐酸酒精分化数秒,自来水冲洗,0.6%氨水返蓝,流水冲洗;切片入伊红染液中染色1~3min;再将切片依次放入95%酒精I 5min→95%酒精II 5min→无水乙醇I 5min→无水乙醇II 5min→二甲苯I 5min→二甲苯II 5min中脱水透明,将切片从二甲苯拿出来稍晾干,中性树胶封片。
结果:如图9A所示,STC1过表达的肿瘤细胞体积显著大于野生型细胞,同样经过60mg/Kg的替莫唑胺治疗,STC1过表达的肿瘤细胞体积仍显著大于野生型细胞组。当敲降STC1基因表达,如图9B所示,与野生型细胞相比较,肿瘤细胞体积显著减小而且经过60mg/kg的替莫唑胺治疗后,肿瘤体积显著小于野生型细胞组。
结合上述结果,STC1在胶质瘤中显著高表达,且高级别胶质瘤的STC1表达量高于低级别胶质瘤;另一方面,STC1高表达可以用于胶质瘤的辅助诊断、化疗疗效预测及预后判断,并在临床样本中对以上结论进行了验证。
以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (10)
1.胶质瘤标记物STC1在制备胶质瘤辅助诊断、疗效预测及预后判断试剂中的应用。
2.根据权利要求1所述的应用,其特征在于,所述试剂包括特异性结合STC1蛋白的抗体或蛋白质芯片,特异性检测STC1 mRNA的引物、探针、核酸芯片。
3.根据权利要求1所述的应用,其特征在于,所述胶质瘤为新鲜的、冷冻的或石蜡固定包埋的组织。
4.STC1表达抑制剂在制备抑制胶质瘤细胞增殖药物中的应用。
5.根据权利要求4所述的应用,其特征在于,所述抑制剂选自以STC1基因或其转录本为靶序列、且能够抑制其蛋白表达或基因转录的反义核酸;或能表达或形成所述反义核酸的构建物。
6.根据权利要求5所述的应用,其特征在于,所述抑制剂包括miRNA、siRNA、dsRNA或shRNA;或包含所述miRNA、siRNA、dsRNA或shRNA的过表达质粒载体或慢病毒。
7.根据权利要4所述的应用,其特征在于,所述抑制胶质瘤细胞增殖药物包括胶质瘤化疗药物和/或药学上可接受的载体。
8.根据权利要7所述的应用,其特征在于,所述胶质瘤化疗药物包括替莫唑胺。
9.胶质瘤标记物STC1在体外筛选治疗和/或预防胶质瘤药物、预防胶质瘤转移、预防胶质瘤侵袭药物中的应用。
10.根据权利要求9所述的应用,其特征在于,所述药物能够抑制STC1高表达。
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2015133771A1 (ko) * | 2014-03-06 | 2015-09-11 | 주식회사 노바셀테크놀로지 | 신규 신경교종의 바이오마커 및 그의 용도 |
CN108469522A (zh) * | 2017-02-23 | 2018-08-31 | 北京师范大学 | 神经胶质瘤的蛋白标志物及其用途 |
WO2019057913A1 (en) * | 2017-09-21 | 2019-03-28 | Fundación Para La Investigación Médica Aplicada | METHOD AND KITS FOR THE PROGNOSIS OF SQUAMOUS CARCINOMA PULMONARY (SCC) |
WO2019057919A1 (en) * | 2017-09-21 | 2019-03-28 | Fundación Para La Investigación Médica Aplicada | METHODS AND KITS FOR THE PROGNOSIS OF LUNG ADENOCARCINOMA |
KR20200070890A (ko) * | 2018-12-10 | 2020-06-18 | 고려대학교 산학협력단 | 상처치료제 스크리닝용 조성물 및 이를 이용한 상처 치료제 스크리닝 방법 |
KR20220170379A (ko) * | 2021-06-22 | 2022-12-29 | 동아대학교 산학협력단 | 암의 진단용 신규한 바이오마커 및 이의 용도 |
WO2022270926A1 (ko) * | 2021-06-22 | 2022-12-29 | 동아대학교 산학협력단 | 다양한 암의 진단, 전이 또는 예후 예측용 신규한 바이오마커 및 이의 용도 |
-
2023
- 2023-06-19 CN CN202310733743.9A patent/CN116718784A/zh active Pending
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2015133771A1 (ko) * | 2014-03-06 | 2015-09-11 | 주식회사 노바셀테크놀로지 | 신규 신경교종의 바이오마커 및 그의 용도 |
CN108469522A (zh) * | 2017-02-23 | 2018-08-31 | 北京师范大学 | 神经胶质瘤的蛋白标志物及其用途 |
WO2019057913A1 (en) * | 2017-09-21 | 2019-03-28 | Fundación Para La Investigación Médica Aplicada | METHOD AND KITS FOR THE PROGNOSIS OF SQUAMOUS CARCINOMA PULMONARY (SCC) |
WO2019057919A1 (en) * | 2017-09-21 | 2019-03-28 | Fundación Para La Investigación Médica Aplicada | METHODS AND KITS FOR THE PROGNOSIS OF LUNG ADENOCARCINOMA |
KR20200070890A (ko) * | 2018-12-10 | 2020-06-18 | 고려대학교 산학협력단 | 상처치료제 스크리닝용 조성물 및 이를 이용한 상처 치료제 스크리닝 방법 |
KR20220170379A (ko) * | 2021-06-22 | 2022-12-29 | 동아대학교 산학협력단 | 암의 진단용 신규한 바이오마커 및 이의 용도 |
WO2022270926A1 (ko) * | 2021-06-22 | 2022-12-29 | 동아대학교 산학협력단 | 다양한 암의 진단, 전이 또는 예후 예측용 신규한 바이오마커 및 이의 용도 |
Non-Patent Citations (5)
Title |
---|
JINGYUAN SU等: "Stanniocalcin-1, a new biomarker of glioma progression, is associated with prognosis of patients", TUMOR BIOLOGY, vol. 36, pages 6333 - 6339, XP036218533, DOI: 10.1007/s13277-015-3319-0 * |
JUNICHI SAKATA等: "MicroRNA regulating stanniocalcin-1 is a metastasis and dissemination promoting factor in glioblastoma", JOURNAL OF NEURO-ONCOLOGY, vol. 142, pages 241 - 251, XP036750901, DOI: 10.1007/s11060-019-03113-2 * |
WEIJIAN LUO等: "Stanniocalcin 1 is a prognostic biomarker in glioma", ONCOLOGY LETTERS, vol. 20, pages 2248 - 2256 * |
YAN XIONG等: "STC1 regulates glioblastoma migration and invasion via the TGF‑β/SMAD4 signaling pathway", MOLECULAR MEDICINE REPORTS, vol. 20, pages 3055 - 3064 * |
仲丽芸等主编: "脑胶质瘤百科问答", 30 September 2021, 中国协和医科大学出版社, pages: 86 * |
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