不同分子标记物在甲状腺乳头状癌中的表达改变及其诊断价值

目的:探讨各种分子标记物在甲状腺乳头状癌(PTC)中表达变化及其对PTC的诊断价值。方法:用免疫组化法检测85例PTC及115例良性甲状腺结节(BTN)患者的术后病理标本中TPO、CK19、Galectin-3、HBME-1及Ki-67的表达,用受试者工作特征曲线(ROC)及二元Logistic回归方程评估各标记物及其联合检测对PTC的诊断价值。结果:与BTN组织比较,PTC组织中CK19、Galectin-3、HBME-1、Ki-67的表达水平均明显升高,而TPO的表达水平明显降低(均P0.05)。单独检测时,TPO的ROC曲线下面积(AUC)值最大,为0.943;联合检测时,5种分子标记物联合检测的AUC值最大,为0.996,但与其他任何包含Galectin-3与TPO两种标记物的组合比较,差异均无统计学意义(均P0.05)。结论:多种分子标记物在PTC中的表达有改变,其中Galectin-3及TPO联合检测可能对提高PTC的诊断有较高的价值。     

galectin-3、HBME-1、CK19及RET在甲状腺肿瘤中的表达情况及其临床价值

目的分析半乳糖凝集素-3(galectin-3)、人骨髓内皮细胞-1(HBME-1)、细胞角蛋白(CK)19及RET在甲状腺良、恶性肿瘤中的表达情况,探讨其在诊断甲状腺肿瘤中的临床价值。方法收集2009～2012年期间我院甲状腺外科经手术切除的131例甲状腺肿瘤患者的临床病理资料,其中甲状腺恶性肿瘤患者45例,甲状腺良性肿瘤患者86例,应用免疫组织化学方法检测galectin-3、HBME-1、CK19及RET在甲状腺良、恶性肿瘤中的表达情况。结果 galectin-3、HBME-1、CK19及RET在45例甲状腺恶性肿瘤患者中的表达阳性率分别为97.8%(44/45)、88.9%(40/45)、100%(45/45)及71.1%(32/45),均明显高于其在86例甲状腺良性疾病患者中的表达阳性率〔分别为9.3%(8/86)、12.8%(11/86)、37.2%(32/86)及8.1%(7/86)〕,差异均有统计学意义(P<0.05)。galectin-3、HBME-1、CK19及RET诊断甲状腺良、恶性疾病的灵敏度分别为97.8%、88.9%、100%及71.1%,特异度分别为90.7%、87.2%、62.8%及91.9%,诊断符合率分别为93.1%、87.8%、75.6%及84.7%。结论 galectin-3、HBME-1、CK19及RET均在甲状腺恶性肿瘤中表达增强,是甲状腺良、恶性疾病病理诊断中有价值的辅助诊断指标,其中galectin-3对甲状腺恶性肿瘤的诊断有较高的参考价值,联合检测可以较大程度上提高甲状腺癌的诊断率。     

甲状腺乳头状癌组织中CXCR7和HBME-1蛋白的表达及意义

探讨甲状腺乳头状癌(PTC)组织中趋化因子受体7(CXCR7)、人骨髓内皮细胞标志物(HBME-1)表达与肿瘤发生发展的关系。选取90例PTC手术后标本(PTC组)和90例甲状腺良性肿瘤组织(良性组),分别采用免疫组织化学染色、Western blot技术检测两组标本中CXCR7蛋白、HBME-1蛋白表达情况,分析不同病灶直径、肿瘤临床分期、淋巴结转移、包膜浸润情况下PTC组织中CXCR7蛋白、HBME-1蛋白阳性表达率。PTC组织中的HBME-1蛋白、CXCR7蛋白阳性表达率分别为71.11%、63.33%,均高于良性组的25.56%、24.44%,差异有统计学意义(P<0.05);经Western blot检测,PTC组织中的CXCR7蛋白、HBME-1蛋白相对表达强度高于良性组,差异有统计学意义(P<0.05);HBME-1蛋白阳性表达率在不同临床分期、是否发生淋巴结转移的PTC肿瘤组织中比较,差异有统计学意义(P<0.05);CXCR7蛋白阳性表达率在是否发生淋巴结转移的PTC肿瘤组织中比较,差异有统计学意义(P<0.05)。PTC组织中CXCR7蛋白、HBME-1蛋白表达增强,可能与肿瘤的发生发展有一定的关系。     

BRAFV600E 突变与乳头状甲状腺癌中央区淋巴结转移的关系

目的：探讨乳头状甲状腺癌（PTC）中BRAF基因突变的临床意义。 方法：选择2年间符合条件的PTC手术患者126例,提取患者石蜡切片中的DNA样本,用荧光PCR法检测BRAF基因V600E（BRAFV600E）的突变情况,分析BRAFV600E突变与PTC患者临床病理因素及中央区淋巴结转移的关系。 结果：126例PTC患者BRAFV600E突变的发生率为69.0%（87/126）。单因素与多因素分析显示,BRAFV600E突变与淋巴结转移有关（P<0.05）;中央区淋巴结转移与年龄、肿瘤大小、肿瘤分期和BRAFV600E突变有关（均P<0.05）。进一步分析显示,肿瘤直径≤10 mm时,BRAFV600E突变与中央区淋巴结转移无关（P>0.05）,肿瘤直径>10 mm时,BRAFV600E突变患者的中央区淋巴结转移率高于突变阴性患者（P<0.05）。 结论：BRAFV600E突变是PTC中央区淋巴结转移风险的独立预测因子;对于术前检测BRAFV600E突变阳性患者,肿瘤直径越大越应该重视中央区淋巴结清扫。     

超声下甲状腺结节内微钙化对良恶性的鉴别及其与甲状腺乳头状癌临床病理特征的关系

目的：探讨甲状腺结节内微钙化对鉴别甲状腺良恶性结节的意义及其与甲状腺乳头状癌（PTC）临床病理特征的关系。 方法：回顾2011年5月—2013年11月期间中南大学湘雅医院手术治疗的743例甲状腺结节患者,分析患者的一般资料、术前彩色超声以及术后病理结果。 结果：743例甲状腺结节中恶性结节137例（18.44%）;甲状腺结节伴微钙化168例,其中恶性结节81例（48.2%）;甲状腺结节伴微钙化者恶性结节的发生率明显高于不伴微钙化者（P<0.05）。微钙化患者中,微钙化密集分布、以实性为主的结节、结节内血流丰富以及伴有淋巴结钙化者甲状腺癌发生率增高（均P<0.05）。PTC伴微钙化的癌结节直径大、甲状腺包膜受侵及颈部淋巴结转移的比例增高（均P<0.05）。 结论：超声下微钙化对甲状腺结节良恶性的鉴别诊断具有重要的临床意义,结合其他超声特征更有助于进一步提高甲状腺癌诊断的准确性。PTC合并微钙化与不良临床病理特征有关。     

CD147 检测对细针穿刺细胞学可疑恶性的甲状腺结节的诊断价值

目的：探讨CD147检测在细针穿刺活检未能明确细胞学诊断的甲状腺结节中的临床应用价值。 方法：选择2013年10月—12月收治的甲状腺结节患者，对术后手术标本进行细针穿刺涂片，用Bethesda甲状腺细胞学报告系统评价腺细胞病理学，选择36例“可疑恶性肿瘤”标本行免疫组化CD147染色分析。CD147阳性者作恶性诊断，结果与手术后石蜡病理结果比较。 结果：CD147检测诊断良性病变19例（52.78%），恶性病变17例（47.22%），术后病理检查诊断良性病变16例（44.44%），恶性病变20例（55.56%）；甲状腺恶性标本中CD147的阳性表达率明显高于良性标本（P<0.05）。CD147的表达与甲状腺结节是否钙化（CD147阳性率89.47%）及淋巴转移（CD147阳性率100%）有关（P<0.05）。细针穿刺细胞学联合CD147免疫组化染色鉴别甲状腺结节良、恶性的准确性和敏感度分别为91.67%和85.0%。 结论：CD147可以作为甲状腺肿瘤细胞学诊断分子生物学标志物，细针穿刺细胞学联合CD147免疫组化分析，有助于提高甲状腺癌的术前检出率。     

BRAF基因突变辅助甲状腺结节诊断为甲状腺乳头状癌的价值研究进展

目的总结BRAF基因突变辅助甲状腺结节诊断为甲状腺乳头状癌(PTC)的价值研究进展。方法收集近年来国内外有关BRAF基因突变及其联合细针穿刺细胞学检查(FNAC)诊断甲状腺良恶性结节、PTC的相关文献并作综述。结果 BRAF基因突变是PTC遗传分子中最常见的基因突变类型,BRAF基因检测联合FNAC可以提高甲状腺良恶性结节特别是PTC诊断的准确性,但BRAF基因突变检测阴性也并不能排除PTC的可能,BRAF基因突变检测对鉴别甲状腺结节良恶性仍存在一定的争议。结论 BRAF基因突变检测在不同类别的甲状腺结节中诊断价值不同,在其突变发生率较高类别的甲状腺结节(可疑的恶性肿瘤、意义不明确的细胞非典型性病变或滤泡性病变结节)中具有较高的诊断价值,而在突变发生率极低类别的甲状腺结节中很大程度上会出现假阴性,尽管如此,BRAF基因检测还是有望成为提高PTC诊断率的高特异性诊断分子标志物。     

甲状腺乳头状癌中BRAF^V600E基因突变的临床意义

目的总结BRAFV600E基因突变对甲状腺乳头状癌（papillary thyroid cancer,PTC）辅助诊断、治疗及预后评估的意义。方法收集近年来国内外有关BRAFV600E基因突变与PTC形成与进展关系的文献并作综述。结果 BRAFV600E基因突变是PTC组织中最常见的突变类型,是促使PTC形成与进展的重要分子改变,其与PTC的高侵袭性及复发,甚至与预后不良均有密切关系。检测甲状腺结节中的BRAFV600E基因突变,不仅能辅助术前细针穿刺细胞学检查（fine needle aspiration cytology,FNAC）对良恶性结节的鉴别诊断,同时能提高FNAC对PTC的诊断准确率,而且对PTC复发危险程度的分级、手术方式及随访计划的制定均具有重要的指导价值。结论 PTC中BRAFV600E基因突变较常见,该基因突变对PTC的诊断及预后评估均具有重要意义。     

甲状腺癌分子生物学的研究进展

甲状腺癌相关基因和肿瘤标志物的研究对于揭示甲状腺癌的发病机制和指导临床诊治均有重要意义。笔者就近年来研究较多且与甲状腺癌密切相关的基因,包括ras,BRAF,Ret/PTC,PAX8/PPARγ,p53和p27;以及肿瘤标志物,包括Gal-3,HBME-1,CK19,MMPs,VEGF,端粒酶和端粒酶逆转录酶等进行综述。     

超声检查联合TSH和Tg评估甲状腺结节良恶性的价值

探讨超声检查联合血清促甲状腺激素(TSH)、甲状腺球蛋白(Tg)判断甲状腺结节良恶性的临床价值。选取2020年6月—2021年12月经病理学检查确诊的甲状腺恶性结节患者94例(恶性组)和同期经病理学检查证实为甲状腺良性结节的患者100例(良性组),通过甲状腺超声检查、血清TSH、Tg水平,对比两组患者超声特征参数、Adler血流分级、TSH和Tg水平,采用受试者工作曲线(ROC)分析超声及血清TSH/Tg鉴别诊断甲状腺良恶性结节的临床价值。结果显示,恶性组的甲状腺结节不规则形态、结节边界不清晰、结节纵横径>1、Adler血流分级(2级+3级)比例均大于良性组(P<0.05);良性组的钙化灶比率显著高于恶性组(P<0.05);鉴别诊断甲状腺良恶性结节的灵敏度甲状腺结节形态分别为63.83%和58.00%,甲状腺结节边界分别为71.28%和67.00%;甲状腺结节钙化灶分别为68.09%和55.00%;甲状腺结节纵横径指标分别为24.47%和93.00%;甲状腺结节低回声分别为80.85%和30.00%;甲状腺结节Adler血流分级分别为82.98%和59.00%;恶性组的血清TSH、TSH/Tg均显著高于良性组(P<0.05);恶性组的Tg值低于良性组(P<0.05);血清TSH/Tg诊断甲状腺良恶性结节的敏感度为69.45%、特异度为70.20%,ROC曲线下面积AUC值为0.746。结果表明,超声特征结合血清TSH/Tg水平对判断甲状腺结节良恶性具有一定的价值。     

甲状腺肿瘤组织中BRAF基因突变及临床意义

目的 探讨甲状腺良恶性肿瘤中BRAF基因点突变及在甲状腺乳头状癌(PTC)中BRAF突变与临床病理学特征之间的关系.方法 应用聚合酶链反应(PCR)技术和基因直接测序法检测104例甲状腺良恶性肿瘤组织中BRAF点突变.结果 在104例标本中,仅在PTC中检测到BRAF点突变,突变率为58.2%(46/79),其他类型的甲状腺良恶性肿瘤中均未检测到BRAF突变.在PTC中BRAF基因突变与肿瘤高分期、腺体外浸润及淋巴结转移呈正相关(P<0.05),而与患者的年龄、性别及肿瘤大小无明显相关(P>0.05).结论 BRAF基因突变是PTC中较常见的遗传学事件,它与肿瘤的分期、腺体外浸润及淋巴结转移具有重要关系.

Abstract：

Objective To investigate the BRAF mutation in benign and malignant thyroid tumors and its relationship with clinicopathological features in papillary thyroid cancer (PTC). Methods Polymerase chain reaction (PCR) and gene sequencing were performed to detect BRAF mutation in 104 cases of benign and malignant thyroid tissues. Results In 104 cases, BRAF mutation was detected in 46 out of 79 PTC cases with a detection rate of 58.2%. BRAF mutation was negative in other kinds of thyroid cancer and benign thyroid diseases. BRAF mutation in PTC was positively correlated with clinical stage, extra-thyroidal extension and lymph nodal metastasis (P<0.05), but not with age, gender and tumor size (P>0.05). Conclusion BRAF mutation was an important and common molecular hallmark of PTC and had a significant correlation with the clinical stage, extra-thyroidal extension and lymph nodal metastasis of PTC.     

Detection of Papillary Thyroid Carcinoma by Analysis of BRAF and RET/PTC1 Mutations in Fine-needle Aspiration Biopsies of Thyroid Nodules

Background  

Activating mutations of the oncogene BRAF or rearrangements of the tyrosine kinase receptor RET are observed in up to 80% of papillary thyroid carcinomas (PTCs). The predominant BRAF V600E mutation has not been detected in benign thyroid tissue so far, so consequently, this assumedly pathognomonic alteration is qualified to improve the preoperative diagnosis of PTC.     

Papillary Thyroid Carcinoma Variants

Papillary thyroid carcinomas are the most common thyroid cancers and constitute more than 70% of thyroid malignancies. The most common etiologic factor is radiation, but genetic susceptibility and other factors also contribute to the development of papillary thyroid carcinoma. The most common variants include conventional, follicular variant and tall cell variant. However, many other uncommon variants have been described including oncocytic, columnar cell, diffuse sclerosing and solid forms. Immunohistochemical staining with TTF-1 and thyroglobulin is very useful in confirming the diagnosis of papillary thyroid carcinoma especially in metastatic sites. Markers such as HBME-1 and CITED1 can assist in separating some difficult cases of follicular variants of papillary thyroid carcinomas from follicular adenomas. Molecular studies have shown that the BRAF V600E mutation is found mainly in papillary and anaplastic thyroid carcinomas. Other molecular markers such as HMGA2 and insulin-like growth factor II mRNA binding protein 3 have been used recently as molecular tests to separate papillary thyroid carcinoma and its variants from follicular adenomas and other benign thyroid nodules.     

BRAF in papillary thyroid carcinoma of ovary (struma ovarii)

BACKGROUND: Malignant struma ovarii (MSO) are rare tumors that arise from ectopic thyroid tissue in the ovary, benign struma ovarii (BSO). Most MSO are histologically classified as papillary thyroid carcinomas (PTC). Oncogenic activation of BRAF (35% to 69%), RAS (10%), or RET (5% to 30%) is common in PTC, and the mutations correlate with tumor subtype, patient age, and clinical behavior. In this study, we explored the possible role of these genes in the development of BSO and MSO. DESIGN: Six paraffin-embedded cases of MSO with histopathologic features of PTC (4 follicular variants, 1 classic, and 1 metastasis of a classic) and 9 BSO were identified. BRAF, NRAS, and KRAS mutations were evaluated using a combination of polymerase chain reaction, denaturing high performance liquid chromatography, and automated sequencing. RET alterations were screened by fluorescence in situ hybridization with multicolor probes. Corresponding benign tissues were evaluated when available. RESULTS: BRAF mutations were present in 4 of 6 MSO and none of 9 BSO. The BRAF mutations included V600E (2 cases), K601E, and a novel deletion/substitution TV599-600M. Neither MSO nor BSO contained alterations in NRAS, KRAS, or RET. CONCLUSIONS: The development of MSOs with PTC features is associated with BRAF mutations of the type commonly observed in PTC, suggesting a common pathogenesis for all PTCs regardless of location. In contrast, mutations in the RET/RAS/RAF pathway are not found in BSO. The prognostic significance of BRAF mutation status in MSO remains to be determined.     

乳头状甲状腺癌与BRAF基因相关性的研究进展

鼠类肉瘤滤过性毒菌致癌同源体B1（BRAF）基因突变是甲状腺癌中最常见的遗传学事件,在乳头状甲状腺癌（PTC）中发生率最高。目前研究证明BRAF V600E基因突变与PTC的发生、发展密切相关。并且作为PTC高效的风险预测分子标记物,能够帮助外科医生制定更为个体化的治疗方案。笔者就PTC与BRAF基因相关性的最新研究进展进行综述。     

Epidermal growth factor receptor overexpression is a marker for adverse pathologic features in papillary thyroid carcinoma

Background

Epidermal growth factor receptor (EGFR) overexpression (EGFR-H) is implicated in thyroid carcinoma disease progression; however, the clinicopathologic significance of EGFR-H in tumors that harbor EGFR and/or v-Raf murine sarcoma viral oncogene homolog B1 (BRAF)(V600E) mutations is unknown.

Methods

Tissue microarrays from 81 patients who had undergone thyroidectomy for carcinoma from 2002–2011 were scored for EGFR expression using immunohistochemistry. Somatic mutations in EGFR exons 19 and 21 and BRAF were analyzed. Correlations between the EGFR immunohistochemistry, EGFR, and BRAF(V600E) mutations and the clinicopathologic features were assessed.

Results

EGFR-H was detected in 39.5% of carcinomas (n = 32) from patients with papillary (PTC, 46.2%, n = 18), follicular (29.6%, n = 8), and anaplastic (100.0%, n = 6) but not medullary (0.0%, n = 9) thyroid carcinoma. BRAF(V600E) mutations were identified in 22.2% of the carcinoma cases (n = 18, 15 PTCs and 3 anaplastic thyroid carcinomas). No somatic EGFR mutations were detected in any subtype. On PTC univariate analysis, EGFR-H correlated with increasing stage, extrathyroid extension, tumor capsule invasion, adverse pathologic features (any demonstration of extrathyroid extension, tumor capsule invasion, lymphovascular invasion, lymph node metastasis, and/or distant metastasis), and BRAF(V600E) mutations. On multivariate analysis, EGFR-H correlated with BRAF(V600E) mutations. In BRAF wild-type PTCs, the correlation between EGFR-H and adverse pathologic features approached statistical significance (P = 0.065).

Conclusions

EGFR-H could be an important biomarker for aggressive PTCs, particularly in BRAF wild-type PTCs. Despite EGFR-H in PTC, follicular thyroid carcinoma, and anaplastic thyroid carcinoma by immunohistochemistry, somatic EGFR mutations were absent. Therefore, future investigations of EGFR should consider histologic and immunohistochemical methods, in addition to molecular profiling of thyroid carcinomas. This multimodal approach is particularly important for future clinical trials testing anti-EGFR therapy.