CN104122392A - Ccl15趋化因子在制备筛选甲状腺滤泡癌试剂中的应用 - Google Patents

Ccl15趋化因子在制备筛选甲状腺滤泡癌试剂中的应用 Download PDF

Info

Publication number
CN104122392A
CN104122392A CN201410286443.1A CN201410286443A CN104122392A CN 104122392 A CN104122392 A CN 104122392A CN 201410286443 A CN201410286443 A CN 201410286443A CN 104122392 A CN104122392 A CN 104122392A
Authority
CN
China
Prior art keywords
ccl15
thyroid follicular
thyroid
application
carcinoma
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201410286443.1A
Other languages
English (en)
Other versions
CN104122392B (zh
Inventor
宁光
黄凤娇
王曙
叶蕾
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Ruinjin Hospital Affiliated to Shanghai Jiaotong University School of Medicine Co Ltd
Original Assignee
Ruinjin Hospital Affiliated to Shanghai Jiaotong University School of Medicine Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ruinjin Hospital Affiliated to Shanghai Jiaotong University School of Medicine Co Ltd filed Critical Ruinjin Hospital Affiliated to Shanghai Jiaotong University School of Medicine Co Ltd
Priority to CN201410286443.1A priority Critical patent/CN104122392B/zh
Publication of CN104122392A publication Critical patent/CN104122392A/zh
Application granted granted Critical
Publication of CN104122392B publication Critical patent/CN104122392B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G01N33/57492—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds localized on the membrane of tumor or cancer cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Cell Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Oncology (AREA)
  • Biotechnology (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Hospice & Palliative Care (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

本发明属于生物医药领域，特别涉及CCL15趋化因子在制备筛选甲状腺滤泡癌试剂中的应用。本发明以确诊为甲状腺滤泡癌的新鲜甲状腺滤泡肿瘤组织标本为研究对象，对其进行了蛋白提取、细胞因子抗体芯片检测、细胞培养、细胞趋化实验和抗体中和实验等，验证了甲状腺滤泡癌中CCL15高表达，而且CCL15高表达组的巨噬细胞浸润密度显著大于CCL15低表达组，FTC培养上清可促进Thp-1单核细胞株的迁移。所以，CCL15可作为潜在的生物标记物，应用于甲状腺滤泡癌的筛选试剂中，同时也为CCL15应用在甲状腺滤泡癌的筛选试剂中提供了实验数据和理论基础。

Description

CCL15趋化因子在制备筛选甲状腺滤泡癌试剂中的应用
技术领域
本发明属于生物医药领域，特别涉及CCL15趋化因子在制备筛选甲状腺滤泡癌试剂中的应用。
背景技术
甲状腺滤泡癌(follicular thyroid carcinoma,FTC)作为分化型甲状腺癌的类型之一，其发病率仅次于甲状腺乳头状癌。大多数分化好的FTC和甲状腺滤泡性腺瘤(follicular adenoma,FA)无论在临床表现还是影像学特点上几乎无法鉴别；在细胞学形态上，细针穿刺(fine needleaspiration,FNA)不能可靠地判断滤泡性肿瘤是否浸润包膜外或侵犯包膜血管，因此不能用于区分滤泡性癌或腺瘤。
同样，在术中行冷冻切片诊断时，也不可能对包膜做彻底检查，因此只有在多取材后才能作出明确诊断。在组织病理学上，这两种肿瘤的实质成分基本相同，唯一能区别的是包膜外和(或)血管侵犯。由于甲状腺滤泡性肿瘤大多为孤立性、有包膜，即使石蜡切片证实包膜外有微小浸润，术中行甲状腺腺叶切除术已能达到治疗目的，预后很好，10年存活率为70％～100％。而广泛浸润的甲状腺滤泡癌通常在大体和光镜下容易证实浸润性生长，肿瘤可侵犯到甲状腺外，手术切除后易复发和血道转移，预后较差，10年存活率仅25％～45％。
但是，对于冷冻切片病理报告诊断为“甲状腺滤泡性肿瘤，须等石蜡切片确定肿瘤有无包膜和/或血管侵犯”的患者，面临着二次手术的可能。正是由于甲状腺滤泡癌和甲状腺腺瘤的鉴别困难，外科医生难以制定合适的手术方案。
因此，如果能对甲状腺滤泡癌进行有效地检测和筛选，对病人的及时治疗有着至关重要的作用。
发明内容
本发明的目的是研究CCL15细胞因子在甲状腺滤泡癌(FTC)中的作用，为临床上治疗和检测甲状腺滤泡癌提供理论依据和实验数据。
甲状腺滤泡癌的病理特征是显示侵袭性的滤泡细胞肿瘤，同时缺少乳头状癌诊断性的核特征。肿瘤微环境是指除肿瘤实质细胞以外的其他所有细胞总称，其中数量最多的是经血液运输迁移至肿瘤部位的巨噬细胞，为肿瘤微环境中的主要成分。巨噬细胞来源于骨髓CD34+祖细胞，首先在骨髓中增殖分裂，再以单核细胞的形式经血液运输迁移。
本研究发现，巨噬细胞浸润散在分布于甲状腺滤泡性肿瘤的滤泡间质，在癌旁正常甲状腺组织中几乎不存在。在甲状腺滤泡癌中，巨噬细胞的浸润密度显著高于其在甲状腺瘤中的密度。甲状腺滤泡癌中的巨噬细胞表型倾向于M2型。
在本课题中，我们分析了甲状腺滤泡癌(FTC)和甲状腺滤泡性腺瘤(FA)的细胞因子谱，二者均分泌MCP-1、CSF-1、VEGF等细胞因子，其中CCL15的表达差异最为显著。
CCL15是巨噬细胞炎症蛋白家族的一员，对单核细胞、T淋巴细胞及内皮细胞具有趋化作用。在我们的研究中，甲状腺滤泡癌细胞上CCL15的表达显著高于其在FA中的表达，并与巨噬细胞的浸润密度显著相关。在细胞功能实验中，CCL15中和抗体可抑制FTC133的上清对Thp-1的趋化作用。结合组织芯片上FTC浸润的巨噬细胞倾向于M2表型，显示甲状腺滤泡癌细胞通过分泌CCL15招募血液中的单核细胞迁移至肿瘤中心，促进肿瘤的发生发展。
基于CCL15在FTC中的高表达，CCL15可作为潜在的生物标记物，应用于FTC患者血清或甲状腺结节细针穿刺细胞盥洗液的检验，为鉴别FTC奠定一定的理论基础。
与现有技术相比，本发明的有益效果在于：1、本发明验证了甲状腺滤泡癌中CCL15高表达，甲状腺滤泡癌细胞通过分泌CCL15招募血液中的单核细胞迁移至肿瘤中心，促进肿瘤的发生发展。所以，CCL15可作为潜在的生物标记物，应用于甲状腺滤泡癌的检查。2、本发明为CCL15应用在甲状腺滤泡癌的筛选试剂中提供了实验数据和理论基础。
附图说明
图1为CCL15在甲状腺滤泡癌中的表达图,其中，图中的A为CCL15高表达组，图中的B为CCL15低表达组。
图2为CCL15对Thp-1细胞株的趋化作用图。
具体实施方式
下面结合实施例，对本发明作进一步说明：
实施例1
将确诊为甲状腺滤泡癌的新鲜手术组织标本，迅速放入组织冻存管，保存于液氮罐中备用。
1、甲状腺滤泡癌分泌细胞因子谱分析
本组实验中，我们提取了FTC和FA的液氮冷冻标本的蛋白，进行细胞因子谱分析。购自RayBio公司的人细胞因子抗体芯片G5含80种细胞因子。共对6个样品进行了芯片分析，4例来自FTC，2例FA。从细胞因子芯片原始检测图中可以看到无论是甲状腺滤泡癌还是甲状腺腺瘤，都表达CCL2、CSF-1、VEGF等趋化因子，但是表达差异最显著的是巨噬细胞炎症蛋白1δ(macrophage inflammatory protein1δ,MIP-1δ)，又名CCL15(chemokine C-C motif ligand15,CCL15)。
2、CCL15在FTC中的表达
CCL15是一种单核细胞趋化因子。在观察到FTC和FA细胞因子谱中CCL15的表达差异后，我们在甲状腺滤泡性肿瘤组织芯片上进行免疫组化染色验证发现，76.7％的FTC病例高表达CCL15,具体如图1所述，图中的A为CCL15高表达组，图中的B为CCL15低表达组。此外,通过分析FTC和FA中CCL15的表达与CD68阳性的巨噬细胞浸润密度的关系发现，CCL15高表达组的巨噬细胞浸润密度显著大于CCL15低表达组。
3、CCL15促进甲状腺滤泡癌细胞对巨噬细胞的招募
基于上述CCL15高表达与巨噬细胞的密度显著相关，我们进一步验证了FTC细胞是否通过分泌CCL15招募单核细胞。FTC133是人的甲状腺滤泡癌细胞株，Thp-1是人的单核细胞株，细胞表面表达CCR1受体。我们收集FTC133的条件培养基(conditional medium,CM)，与Thp-1共培养24h后，观察Thp-1生物学行为的改变。如图2所示，与空白对照组相比FTC133的细胞培养上清能促进Thp-1细胞的迁移。在FTC133-CM的刺激下，Thp-1穿过细胞小室上层粘附在鼠尾胶上，说明FTC133可以通过其分泌的细胞因子直接作用于单核细胞。
综上所述，甲状腺滤泡癌中CCL15高表达，而且CCL15高表达组的巨噬细胞浸润密度显著大于CCL15低表达组，FTC培养上清可促进Thp-1单核细胞株的迁移。所以，CCL15可作为潜在的生物标记物，应用于甲状腺滤泡癌的筛选试剂中，同时也为CCL15应用在甲状腺滤泡癌的筛选试剂中提供了实验数据和理论基础。

Claims (1)

1.CCL15趋化因子在制备筛选甲状腺滤泡癌试剂中的应用。
CN201410286443.1A 2014-06-24 2014-06-24 检测ccl15趋化因子的试剂在制备筛选甲状腺滤泡癌试剂中的应用 Active CN104122392B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201410286443.1A CN104122392B (zh) 2014-06-24 2014-06-24 检测ccl15趋化因子的试剂在制备筛选甲状腺滤泡癌试剂中的应用

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201410286443.1A CN104122392B (zh) 2014-06-24 2014-06-24 检测ccl15趋化因子的试剂在制备筛选甲状腺滤泡癌试剂中的应用

Publications (2)

Publication Number Publication Date
CN104122392A true CN104122392A (zh) 2014-10-29
CN104122392B CN104122392B (zh) 2016-05-25

Family

ID=51767882

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201410286443.1A Active CN104122392B (zh) 2014-06-24 2014-06-24 检测ccl15趋化因子的试剂在制备筛选甲状腺滤泡癌试剂中的应用

Country Status (1)

Country Link
CN (1) CN104122392B (zh)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1668614A (zh) * 2002-07-18 2005-09-14 辉瑞产品公司 用作ccr1趋化因子受体拮抗剂双环哌啶衍生物
CN102746404A (zh) * 2004-11-12 2012-10-24 赞科股份有限公司 对FcRn的结合被改变的Fc变体

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1668614A (zh) * 2002-07-18 2005-09-14 辉瑞产品公司 用作ccr1趋化因子受体拮抗剂双环哌啶衍生物
CN102746404A (zh) * 2004-11-12 2012-10-24 赞科股份有限公司 对FcRn的结合被改变的Fc变体

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DAGMARA RUSINEK，ET AL.: "Gene expression profile of human thyroid cancer in relation to its mutational status", 《JOURNAL OF MOLECULAR ENDOCRINOLOGY》 *
DAGMARA RUSINEK，ET AL.: "Gene expression profile of human thyroid cancer in relation to its mutational status", 《JOURNAL OF MOLECULAR ENDOCRINOLOGY》, vol. 47, no. 3, 2 November 2011 (2011-11-02) *
弓雪莲 等: "CC趋化因子受体5在滤泡状甲状腺癌中的表达", 《第二军医大学学报》 *
李悦国 等: "趋化因子 CCL15 在肝细胞肝癌组织中的表达及其临床意义", 《中国肿瘤临床》 *

Also Published As

Publication number Publication date
CN104122392B (zh) 2016-05-25

Similar Documents

Publication Publication Date Title
Balic et al. Circulating tumor cells: from bench to bedside
Zhang et al. Circulating tumor cells in hepatocellular carcinoma: detection techniques, clinical implications, and future perspectives
Zhang et al. Matrix metalloproteinase-9 as a prognostic factor in gastric cancer: a meta-analysis
Hou et al. Circulating tumor cells as a window on metastasis biology in lung cancer
Kolostova et al. Circulating tumor cells in localized prostate cancer: isolation, cultivation in vitro and relationship to T-stage and Gleason score
Wang et al. Histologic assessment of tumor-associated CD45+ cell numbers is an independent predictor of prognosis in small cell lung cancer
Flaig et al. Detection of circulating tumor cells in metastatic and clinically localized urothelial carcinoma
Feng et al. Circulating tumor cells in the early detection of human cancers
Rossi et al. Noninvasive follicular thyroid neoplasm with papillary‐like nuclear features in the pediatric age group
Wang et al. Cultured circulating tumor cells and their derived xenografts for personalized oncology
CN111665357A (zh) 结外鼻型nk/t细胞淋巴瘤预后标志物及其应用和预后预测模型及其构建方法
Serrano et al. Detection of circulating tumor Cells in the context of treatment: Prognostic value in breast cancer patients
Kiss et al. Correlation between disease stage and the presence of viable circulating tumor cells in endometrial cancer
Jaeschke et al. Cancer-associated fibroblasts of the prostate promote a compliant and more invasive phenotype in benign prostate epithelial cells
Ren et al. Malignant characteristics of circulating tumor cells and corresponding primary tumor in a patient with esophageal squamous cell carcinoma before and after surgery
Hu et al. Enrichment and detection of circulating tumor cells by immunomagnetic beads and flow cytometry
Wong et al. Levels of a subpopulation of platelets, but not circulating endothelial cells, predict early treatment failure in prostate cancer patients after prostatectomy
Suvilesh et al. Preclinical models to study patient-derived circulating tumor cells and metastasis
CN103361312A (zh) 一种甲状腺乳头状癌组织中巨噬细胞的分离方法
Andalib et al. CCR3, CCR4, CCR5, and CXCR3 expression in peripheral blood CD4+ lymphocytes in gastric cancer patients
CN104122392B (zh) 检测ccl15趋化因子的试剂在制备筛选甲状腺滤泡癌试剂中的应用
Röbeck et al. Multiplex protein analysis and ensemble machine learning methods of fine needle aspirates from prostate cancer patients reveal potential diagnostic signatures associated with tumour grade
Chatziavramidis et al. Brush cytology with immunocytochemical evaluation of VEGF expression versus biopsy in clinically precancerous laryngeal lesions: can we diagnose laryngeal cancer only with brush cytology?
Kolostova et al. Circulating tumor cells as an auxiliary diagnostic tool in surgery
Hoshi et al. Discriminant model for cytologic distinction of large cell neuroendocrine carcinoma from small cell carcinoma of the lung

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant