CN107937529A - 胶质瘤诊断标志物hsa_circ_0135404及应用 - Google Patents
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Abstract
本发明属于生物技术领域，公开了一种胶质瘤诊断标志物hsa_circ_0135404及应用。本发明通过提取胶质瘤患者血清外泌体，并对其中hsa_circ_0135404的含量进行实时荧光定量PCR分析，发现胶质瘤患者血清外泌体中hsa_circ_0135404表达水平明显上升(p＝0.0033)，该发现对临床用于胶质瘤的早期诊断具有很高价值，AUC＝0.879，p<0.0001，其敏感度和特异度分别为89.7％和75％。
Description
技术领域
本发明属于生物技术领域，涉及一种用于胶质瘤诊断的血清外泌体circRNA标志物、以及检测该标志物的试剂用于制备胶质瘤诊断制剂的应用、还有试剂盒。
背景技术
胶质瘤是颅内发病率最高的恶性肿瘤，占颅内肿瘤的40.49％，虽然临床上采用手术、化疗结合的方法进行治疗，但由于其浸润性，对化疗药物低敏感性等原因，经常导致患者术后复发，严重威胁人类生命健康。而且一旦确诊，大部分都为胶质瘤中晚期，术后生存率不高。另外，在WHO3-4级即高级别脑胶质瘤患者中，预后较差，平均生存寿命不超过五年。因此，寻找胶质瘤诊断标志物对患者进行诊断分析，尽早确诊病情，并相应地选择合理的后续治疗方案，提高生存率，是神经科学领域亟待解决的研究任务。
circRNA是一类广泛且多样地存在于哺乳动物细胞中、具有调控基因表达作用的内源性非编码RNA分子，具有共价闭合的环形结构，广泛存在于各种细胞中，也是继microRNA(miRNA)后RNA家族的最新研究热点。近年来，随着深度测序技术的广泛应用和生物物理和信息学技术的快速发展，人们发现人类许多外显子的转录本可被非线性地反向剪接或通过基因重排而形成circRNA，且它们在所有剪接转录本中占了相当大的比例。目前，由于其具有丰富性、稳定性、高保守性和时空特异性等特点，在肿瘤诊断标志物方面正发挥越来越大的作用。
发明内容
本发明的第一个目的是：提供一种用于胶质瘤诊断的血清外泌体circRNA标志物。
主要内容包括：一种用于胶质瘤诊断的血清外泌体circRNA标志物hsa_circ_0135404，其序列如SEQ NO:1所示。
本发明的第二个目的是，提供检测所述的circRNA标志物在血清外泌体中表达量的试剂在制备胶质瘤诊断制剂中的应用。
本发明的第三个目的是，提供一种胶质瘤诊断试剂盒，其能够测定血清外泌体中的hsa_circ_0135404的含量。
所述的胶质瘤诊断试剂盒含有检测hsa_circ_0135404含量的PCR引物。优选引物的序列如SEQ NO:2和3所示。
所述的胶质瘤诊断试剂盒，除hsa_circ_0135404的引物外，还含有从血清中提取外泌体、由外泌体中提取RNA并进行逆转录及荧光定量PCR的所有试剂。包括：(1)提取血清外泌体所需试剂：Total Exosome Isolation Reagent(from serum)，可由Invitrogen公司购得，货号4478360；(2)提取外泌体RNA所需试剂：Trizol试剂、三氯甲烷、异丙醇、75％乙醇、无酶水；(3)逆转录所需试剂：随机引物(Random Primer)、无酶水、5×逆转录缓冲液、三磷酸碱基脱氧核苷酸、RNA酶抑制剂、MMLV逆转录酶；(4)荧光定量PCR所需试剂：hsa_circ_0135404上下游引物、GAPDH内参上下游引物、SYBR染料、无酶水。
本发明的有益效果在于：首次发现hsa_circ_0135404这种血清外泌体中的环状RNA，并发现其对胶质瘤具有较高的诊断价值；通过血清circRNA标志物和诊断试剂盒的研制和应用，可以使得胶质瘤的诊断更加方便易行，为临床医生快速准确掌握患者病情，为提高临床治疗效果奠定基础，并为发现具有潜在治疗价值的新型小分子药物靶标提供帮助。
附图说明
图1为健康组与胶质瘤组血清外泌体中CircRNA表达水平比较。
图2为健康组与胶质瘤组血清外泌体中CircRNA表达水平ROC曲线。
具体实施方式
一、研究对象
病例组为在湖南省肿瘤医院收集的29例病例，均经病理组织学确诊，采血前未经过手术和放化疗。对照组为同期进行社区疾病筛查的健康个体12例，按性别和年龄(±5岁)与病例进行频数匹配。用于研究的样本为同期收取，采样、分装、保存条件均一，通过对样品资料的整理。
二、研究方法
(1)血清制备：使用EDTA抗凝管采肘静脉血5ml，采血后轻轻摇动抗凝管使抗凝剂与血液混匀，4℃静止24小时后3000g常温离心5分钟。用微量移液器抽取200μl上层血清分装至新的600μl离心管中，-80℃保存备用。由实验员每天定时记录冰箱温度。
(2)血清外泌体的制备：取上述血清200μl于2000g常温离心30分钟，用微量移液器抽取上层清液至新的600μl离心管，加入40μl外泌体提取试剂(Total Exosome IsolationReagent(from serum)，货号4478360，Invitrogen公司)轻轻上下颠倒摇匀，4℃孵化45分钟。孵化结束后10000g常温离心10分钟，弃掉上清液，于沉淀中加入200μl Trizol(MRC公司)使沉淀重悬，将悬液移至新的1.5mltube管，补Trizol至1ml。
(3)外泌体中RNA的抽提：将上述重悬液于冰上静止裂解15分钟。裂解结束后4℃12000rpm离心10min，上清液移至新的tube管。加氯仿200μl/ml Trizol于Tube中，用手震荡15-30s，冰上放置5min，4℃12000rpm离心15min；小心取上层水相入新tube中，加入预冷的异丙醇0.5ml/mlTrizol混匀，-20℃冰箱静置20min，4℃12000rpm离心10min；弃上清，加入75％DEPC水稀释的乙醇1-2ml混匀，4℃7500rpm离心5min，尽量弃上清，室温干燥5-10min，加入DEPC水10μl溶解RNA。-80℃保存。
(4)cDNA制备：按照RNA逆转录试剂盒(Thermo公司)说明书进行逆转录反应。反应总体积为20μl(总RNA10μl，Random primer1μl,核酸酶自由水1μl，5×Reaction Buffer 4μl，RI 1μl，RT 1μl和10mM dNTP 2μl)。
| 成分 | 剂量/管 |
| 随机逆转录引物(1μM) | 1μl |
| RNA样本 | 10ul |
| 无酶水 | To 12μl |
逆转录第一步条件：65℃5分钟
| 成分 | 剂量/管 |
| 5×逆转录缓冲液 | 4μl |
| 三磷酸碱基脱氧核苷酸(10mM) | 2μl |
| RNA酶抑制剂(40U/μl) | 1μl |
| MMLV逆转录酶(200U/μl) | 1μl |
| 第一步逆转录的产物 | 12μl |
| 总体积 | 20μl |
逆转录第二步程序：25℃5分钟，42℃60分钟，70℃5分钟。
(5)实时荧光定量PCR：广州吉赛生物工程有限公司合成的特异性引物进行实时定量PCR：先将逆转录产物稀释10倍，混匀。20μL反应体系如下：
| 成分 | 剂量/管 |
| SYBR Premix Ex Taq | 10μl |
| 特异性引物(10μM) | 0.5μl |
| cDNA产物(稀释后) | 5μl |
| 无酶水 | To 20μl |
表1引物序列表
实时荧光定量PCR反应程序：95℃3分钟,40个循环，95℃10秒，60℃30秒。
(6)数据分析：：用2-ΔΔCT表示对于对照组变化的倍数，其中△CT＝CT样本–CT内参，△△CT＝△CT病例–△CT对照。本实验数据采用相对定量的分析方法，GAPDH作为内参基因，数据利用软件GraphPad Prism及SPSS 17.0进行分析。
用于胶质瘤诊断的试剂盒包括如表1所示的引物探针，可由广州吉赛公司购得；还可以有相应PCR技术所需的常用试剂，如：Taq酶，逆转录酶，缓冲液，dNTPs，MgCl2和DEPC水等，这些都是本领域技术人员熟知的；另外还可以含有标准品和/或对照品。
三、研究结果
病例组血清hsa_circ_0135404表达水平较正常组显著上调(p＝0.0033)。具体数据如图1所示。
ROC曲线分析显示，hsa_circ_0135404作为生物标记物对胶质瘤具有较高诊断价值(AUC＝0.879，p<0.0001，敏感度和特异度分别为89.7％和75％)。详细结果见图2。
序列表
<110> 中南大学湘雅医院
<120> 胶质瘤诊断标志物hsa_circ_0135404及应用
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<213> 未知(Unknown)
<400> 5
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Claims (6)
1.一种胶质瘤诊断标志物hsa_circ_0135404，其序列如SEQ NO:1所示。
2.检测权利要求1所述的标志物在血清外泌体中表达量的试剂在制备胶质瘤诊断制剂中的应用。
3.一种胶质瘤诊断试剂盒，其特征在于，能够测定血清外泌体中的hsa_circ_0135404的含量。
4.根据权利要求3所述的胶质瘤诊断试剂盒，其特征在于，含有检测hsa_circ_0135404含量的PCR引物。
5.根据权利要求4所述的胶质瘤诊断试剂盒，其特征在于，引物的序列如SEQ NO:2和3所示。
6.根据权利要求3或4或5所述的胶质瘤诊断试剂盒，其特征在于，除hsa_circ_0135404的引物外，还含有从血清中提取外泌体、由外泌体中提取RNA并进行逆转录及荧光定量PCR的所有试剂。
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| CN108660255A (zh) * | 2018-05-31 | 2018-10-16 | 广州医科大学附属第五医院 | 一种用于诊断rsv感染的环状rna及其应用 |
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Non-Patent Citations (2)
| Title |
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| JESSICA C.ET AL.: "Arlm1 is a male-specific modifier of astrocytoma resistance on mouse Chr12", 《NEURO-ONCOLOGY》 * |
| JULIA SALZMAN ET AL.: "Cell-type specific features of circular RNA expression", 《PLOS GENET》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108660255A (zh) * | 2018-05-31 | 2018-10-16 | 广州医科大学附属第五医院 | 一种用于诊断rsv感染的环状rna及其应用 |
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