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Immunohistochemical Investigation of the Proliferative Activity of Odontogenic Cysts and Tumors. 牙源性囊肿和肿瘤增殖活性的免疫组化研究
IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2024-11-21 DOI: 10.1097/PAI.0000000000001240
Manal A Alsheddi

Odontogenic cysts and tumors exhibit a broad spectrum of biological characteristics. Despite recent advances in understanding the complex nature of these lesions, relatively less is known about the molecular markers involved in key pathogenic steps, such as proliferation and differentiation. This study aimed to elucidate the expression patterns of p63 and Ki-67 in odontogenic lesions, which may influence the management strategies. Forty-two specimens from the archives of the Histopathology Laboratory, including conventional ameloblastoma, unicystic ameloblastoma, odontogenic keratocysts, dentigerous cysts, and orthokeratinized cysts, were analyzed. Immunohistochemistry was performed using antibodies against p63 and Ki-67. Digital image analysis was performed using an Aperio slide scanner and QuPath software. Ki-67 levels were higher in odontogenic keratocysts (OKCs), indicating a greater proliferative index, whereas p63 expression was significantly higher in ameloblastomas and OKCs than in dentigerous cysts. No significant difference in p63 expression was observed between the ameloblastoma types. The results revealed variable Ki-67 and p63 expression in the odontogenic epithelium of the investigated odontogenic lesions, suggesting their potential roles in the biological behavior and aggressiveness of these lesions. This study highlights the differential expression pattern of Ki-67 and p63 and their potential involvement in the pathogenesis of these rare lesions. In addition, the study reinforces the need for more comprehensive molecular analyses using a larger sample. The results contribute to a better understanding of the complex nature of these lesions, which may facilitate improving the management options of odontogenic cysts and tumors.

牙源性囊肿和肿瘤具有广泛的生物学特征。尽管近年来人们对这些病变的复杂性有了更深入的了解,但对参与关键致病步骤(如增殖和分化)的分子标志物却知之甚少。本研究旨在阐明 p63 和 Ki-67 在牙源性病变中的表达模式,这可能会影响治疗策略。研究分析了组织病理学实验室档案中的 42 份标本,包括传统的釉母细胞瘤、单囊性釉母细胞瘤、牙源性角化囊肿、牙源性囊肿和正角化囊肿。使用 p63 和 Ki-67 抗体进行免疫组化。使用 Aperio 幻灯片扫描仪和 QuPath 软件进行数字图像分析。牙源性角化囊肿(OKCs)的 Ki-67 水平较高,表明其增殖指数较高,而牙釉质母细胞瘤和 OKCs 的 p63 表达明显高于牙源性角化囊肿。在不同类型的成釉细胞瘤中,p63的表达没有明显差异。研究结果表明,在所调查的牙源性病变中,Ki-67 和 p63 在牙源性上皮细胞中的表达各不相同,这表明它们在这些病变的生物学行为和侵袭性中发挥着潜在的作用。本研究强调了 Ki-67 和 p63 的不同表达模式及其在这些罕见病变发病机制中的潜在作用。此外,该研究还强调了使用更大样本进行更全面分子分析的必要性。研究结果有助于更好地了解这些病变的复杂性,从而有助于改进牙源性囊肿和肿瘤的治疗方案。
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引用次数: 0
Expression of ALKBH5 in Odontogenic Lesions. ALKBH5 在牙源性病变中的表达
IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2024-11-11 DOI: 10.1097/PAI.0000000000001233
Chatchaphan Udompatanakorn, Worawan Sriphongphankul, Patrayu Taebunpakul

N6-methyladenosine (m6A) is the most abundant epigenetic RNA modification in eukaryotes and plays a role in various cancers in humans. This m6A modification is regulated by m6A writers, erasers, and readers. One of the m6A erasers is α-ketoglutarate-dependent dioxygenase homolog 5 (ALKBH5). Previous studies have suggested that ALKBH5 is involved in the pathogenesis of head and neck squamous cell carcinoma. However, the role of ALKBH5 in odontogenic lesions has never been investigated. This study aimed to examine ALKBH5 expression in dental follicles (DFs), dentigerous cysts (DCs), odontogenic keratocyst (OKC), and ameloblastoma (AM) using immunohistochemistry. Six cases of DF, 20 cases of DC and OKC, respectively, and 30 cases of AM were included. The expression patterns, percentage of ALKBH5-positive cells, staining intensities, and immunoreactive scores were examined. ALKBH5 was mainly expressed in the nuclei of the epithelial cells in odontogenic lesions. The percentage of ALKBH5-positive cells was significantly higher in OKC and AM samples compared with DF samples (P < 0.01). The percentage of ALKBH5-positive cells was also higher in OKC and AM samples than in DC samples; however, these results did not show statistical significance (P > 0.05). ALKBH5 cell staining intensities and immunoreactive scores were significantly greater in OKC and AM samples than in DF and DC samples (P < 0.01). Our results suggested that ALKBH5 might play a role in the pathogenesis of odontogenic lesions. Further investigation is needed to elucidate the precise molecular mechanism of the role of ALKBH5 in these diseases.

N6-甲基腺苷(m6A)是真核生物中最丰富的表观遗传 RNA 修饰,在人类的各种癌症中发挥着作用。这种 m6A 修饰受 m6A 写入器、擦除器和阅读器的调控。α-酮戊二酸依赖性二氧合酶同源物 5(ALKBH5)是 m6A 清除剂之一。以前的研究表明,ALKBH5 与头颈部鳞状细胞癌的发病机制有关。然而,ALKBH5 在牙源性病变中的作用却从未被研究过。本研究旨在采用免疫组化方法检测 ALKBH5 在牙泡 (DFs)、牙源性囊肿 (DCs)、牙源性角化囊肿 (OKC) 和釉母细胞瘤 (AM) 中的表达。其中,牙源性囊肿(DF)6 例,牙源性角化囊肿(DC)和牙源性角化囊肿(OKC)20 例,牙源性母细胞瘤(AM)30 例。研究了ALKBH5的表达模式、阳性细胞比例、染色强度和免疫反应评分。ALKBH5主要在牙源性病变的上皮细胞核中表达。与 DF 样本相比,OKC 和 AM 样本中 ALKBH5 阳性细胞的比例明显更高(P < 0.01)。OKC和AM样本中ALKBH5阳性细胞的百分比也高于DC样本,但这些结果均无统计学意义(P > 0.05)。在 OKC 和 AM 样本中,ALKBH5 细胞染色强度和免疫反应得分明显高于 DF 和 DC 样本(P < 0.01)。我们的研究结果表明,ALKBH5可能在牙源性病变的发病机制中发挥作用。要阐明ALKBH5在这些疾病中作用的确切分子机制,还需要进一步的研究。
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引用次数: 0
Immunohistochemical Expression of SATB2 in Malignant Melanomas. 恶性黑色素瘤中 SATB2 的免疫组化表达
IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2024-11-01 Epub Date: 2024-10-10 DOI: 10.1097/PAI.0000000000001229
Rasmus Røge, Birgit Truumees, Søren Nielsen

Accurate diagnosis of cancer of unknown primary (CUP) poses a significant daily challenge for pathologists, necessitating reliable immunohistochemical (IHC) markers. SATB2 is a transcription factor primarily expressed in colorectal neoplasms. This study investigates the IHC expression of SATB2 in malignant melanomas (MM). Using tissue microarrays (TMAs) from Aalborg University Hospital, Denmark, comprising 56 primary and 12 metastatic MMs, we evaluated SATB2 expression through H-scores. We found that 48% of MM cases expressed SATB2, predominantly with weak to moderate staining intensity. Although no significant difference was observed between primary and metastatic MMs, a higher median H-score was noted in metastatic lesions. The results highlight the potential diagnostic pitfall of SATB2 expression in MM and underline the need for careful interpretation.

准确诊断原发灶不明的癌症(CUP)对病理学家来说是一项重大的日常挑战,需要可靠的免疫组化(IHC)标记物。SATB2 是一种主要在结直肠肿瘤中表达的转录因子。本研究调查了 SATB2 在恶性黑色素瘤(MM)中的 IHC 表达。利用丹麦奥尔堡大学医院的组织微阵列(TMA)(包括 56 例原发性和 12 例转移性 MM),我们通过 H 评分评估了 SATB2 的表达。我们发现,48% 的 MM 病例表达 SATB2,主要为弱至中等染色强度。虽然在原发性和转移性 MM 之间没有观察到明显的差异,但在转移性病灶中H-评分的中位数较高。这些结果凸显了SATB2在MM中表达的潜在诊断隐患,并强调了谨慎解读的必要性。
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引用次数: 0
Immunohistochemistry in the Differential Diagnosis of Triple Negative Breast Carcinoma and High-grade Serous Carcinoma: Old and New Markers. 免疫组化在三阴性乳腺癌和高级别浆液性癌鉴别诊断中的应用:新旧标记物。
IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2024-11-01 Epub Date: 2024-11-07 DOI: 10.1097/PAI.0000000000001232
Pragya Virendrakumar Jain, Mariel Molina, Michelle Moh, Erin Bishop, Janet S Rader, Julie M Jorns

Distinction of metastasis to the breast from a breast primary, particularly high-grade triple-negative breast cancer (TNBC), can be challenging due to nonspecific morphology and immunohistochemical (IHC) profiles. Among metastases to the breast, high-grade serous carcinoma (HGSC) of müllerian origin is most likely to be misdiagnosed as TNBC. We assessed breast and müllerian markers on TNBC and HGSC, including keratin 7, keratin 20, GATA3, GCDFP15, mammaglobin, p53, PAX8 (MRQ50 and BC12 clones), TRPS1, SOX10, and WT1. Of 151 TNBC cases, TRPS1 had the highest sensitivity, showing expression in 149 (98.7%) cases, followed by SOX10 (110/151; 72.8%), GATA3 (102/151; 67.5%), GCDFP15 (29/151; 19.2%), and mammaglobin (27/151; 17.9%). PAX8 positivity was seen in 40.4% (61/151) of TNBC via the MRQ50 clone but was negative in all via the BC12 clone. Of 185 HGSC cases, PAX8 via the MRQ50 clone was the most sensitive (179/185; 96.8%), followed by WT1 (171/185; 92.4%) and PAX8 via the BC12 clone (164/185; 88.6%). In addition, TRPS1 positivity was seen in 75 HGSC cases (40.5%). Aberrant p53 patterns were seen in 64.9% (98/151) of TNBC and 94.1% (174/185) of HGSC. TRPS1 positivity in HGSC and PAX8 positivity via the MRQ50 clone in TNBC represent potential pitfalls in assessing high-grade carcinoma for which the differential diagnosis includes TNBC and HGSC. However, with this knowledge, utilization of a panel of breast and müllerian markers, including preferential use of the PAX8 BC12 clone, can facilitate accurate diagnosis.

由于非特异性的形态和免疫组化(IHC)特征,将乳腺转移瘤与乳腺原发癌,尤其是高级别三阴性乳腺癌(TNBC)区分开来具有挑战性。在乳腺转移瘤中,来源于缪勒管的高级别浆液性癌(HGSC)最容易被误诊为 TNBC。我们评估了TNBC和HGSC的乳腺和苗勒氏标记物,包括角蛋白7、角蛋白20、GATA3、GCDFP15、mammaglobin、p53、PAX8(MRQ50和BC12克隆)、TRPS1、SOX10和WT1。在151个TNBC病例中,TRPS1的敏感性最高,在149个病例(98.7%)中均有表达,其次是SOX10(110/151;72.8%)、GATA3(102/151;67.5%)、GCDFP15(29/151;19.2%)和mammaglobin(27/151;17.9%)。40.4%的TNBC(61/151)通过MRQ50克隆检测出PAX8阳性,但通过BC12克隆检测出的PAX8均为阴性。在185例HGSC病例中,通过MRQ50克隆检测的PAX8最敏感(179/185;96.8%),其次是WT1(171/185;92.4%)和通过BC12克隆检测的PAX8(164/185;88.6%)。此外,在 75 例 HGSC(40.5%)中发现了 TRPS1 阳性。64.9%的TNBC(98/151)和94.1%的HGSC(174/185)出现p53模式异常。HGSC中的TRPS1阳性和TNBC中通过MRQ50克隆的PAX8阳性是评估高级别癌的潜在隐患,其鉴别诊断包括TNBC和HGSC。不过,有了这些知识,利用乳腺和苗勒氏管标记物面板,包括优先使用 PAX8 BC12 克隆,可以促进准确诊断。
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引用次数: 0
Expression of B7-H3 and B7-H4 in Gastric Gastrointestinal Stromal Tumors. 胃肠道间质瘤中 B7-H3 和 B7-H4 的表达
IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2024-11-01 Epub Date: 2024-10-02 DOI: 10.1097/PAI.0000000000001227
Kunio Mochizuki, Naoki Oishi, Ippei Tahara, Tomohiro Inoue, Tetsuo Kondo

Gastric gastrointestinal stromal tumors (GISTs) are mesenchymal neoplasms with variable behavior characterized by differentiation toward the interstitial cells of Cajal occurring anywhere in the gastrointestinal stromal tract. The management of GIST was revolutionized by the introduction of imatinib, a KIT inhibitor, which has become the standard first-line treatment for metastatic GIST. However, despite a clinical benefit rate of 80%, the majority of patients with GIST experience disease progression after 2 to 3 years of imatinib therapy. This shows the need for novel treatment approaches for imatinib refractory GISTs. The checkpoint proteins B7-H3 and B7-H4 inhibit the activation and function of T cells by potently suppressing the proliferation, cytokine production, and cytotoxicity of activated T cells, which is a mechanism for immune escape. This study aims to clarify B7-H3 and B7-H4 expression in gastric GISTs using immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). We confirmed B7-H3 expression (H-score ≥50 points) in 92% and B7-H4 expression in 0% of GIST samples. We examined B7-H3 mRNA expression in 3 representative GIST samples, each having their respective immunostained areas detected by RT-PCR. B7-H3 is expressed at a particularly high rate in GISTs. This suggests that B7-H3 might operate as part of an immune checkpoint in GISTs.

胃肠道间质瘤(GIST)是一种间叶肿瘤,其特征是向卡贾尔间质细胞分化,发生在胃肠道间质的任何部位,表现各异。伊马替尼(一种 KIT 抑制剂)的问世彻底改变了 GIST 的治疗方法,它已成为转移性 GIST 的标准一线治疗药物。然而,尽管伊马替尼的临床获益率高达 80%,但大多数 GIST 患者在接受伊马替尼治疗 2 到 3 年后病情仍会恶化。这表明,伊马替尼难治性 GIST 需要新的治疗方法。检查点蛋白B7-H3和B7-H4通过有效抑制活化T细胞的增殖、细胞因子产生和细胞毒性来抑制T细胞的活化和功能,这是一种免疫逃逸机制。本研究旨在利用免疫组化和反转录聚合酶链反应(RT-PCR)明确胃癌 GIST 中 B7-H3 和 B7-H4 的表达。我们在 92% 的 GIST 样本中证实了 B7-H3 的表达(H 评分≥50 分),在 0% 的 GIST 样本中证实了 B7-H4 的表达。我们检测了 3 个具有代表性的 GIST 样本中 B7-H3 mRNA 的表达情况,每个样本都通过 RT-PCR 检测了各自的免疫染色区域。B7-H3 在 GIST 中的表达率特别高。这表明,B7-H3 可能是 GIST 中免疫检查点的一部分。
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引用次数: 0
A Modified Bleaching Method for Multiplex Immunofluorescence Staining of FFPE Tissue Sections. 用于 FFPE 组织切片多重免疫荧光染色的改良漂白法
IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2024-11-01 Epub Date: 2024-10-07 DOI: 10.1097/PAI.0000000000001228
Dan Wang, Alison Cheung, Gordon E Mawdsley, Kela Liu, Yulia Yerofeyeva, Kelsie L Thu, Ju-Yoon Yoon, Martin J Yaffe

Multiplex immunofluorescence (mIF) staining plays an important role in profiling biomarkers and allows investigation of co-relationships between multiple biomarkers in the same tissue section. The Cell DIVE mIF platform (Leica Microsystems) employs an alkaline solution of hydrogen peroxide as a fluorophore inactivation reagent in the sequential staining, imaging, and bleaching protocol for use on FFPE sections. Suboptimal bleaching efficiency, degradation of tissue structure, and loss of antigen immunogenicity occasionally are encountered with the standard bleaching process. To overcome these impediments, we adopted a modified photochemical bleaching method, which utilizes an intense LED light exposure concurrent with the application of hydrogen peroxide. Repeated stain/bleach rounds with different antibodies were performed on breast tissue and other tissue sections. Residual signal after conventional bleaching and the modified technique were compared and tissue integrity and antigen immunogenicity were assessed. The modified technique effectively eliminates fluorescence signal from previous staining rounds and produces consistent results for multiple rounds of staining and imaging. With the modified method, photochemical treatments did not destroy tissue sub-cellular contents, and the tissue antigenicity was well preserved during the entire mIF process. Overall processing time was reduced from 36 to 30 hours in an mIF procedure with 8 rounds. With the conventional method, tissue quality was highly degraded after 8 rounds. The new technique allows reduced turn-around time, provides reliable fluorophore removal in mIF with excellent maintenance of tissue integrity, facilitating studies of the co-localization of multiple biomarkers in tissues of interest.

多重免疫荧光(mIF)染色在分析生物标记物方面发挥着重要作用,可以研究同一组织切片中多种生物标记物之间的相互关系。Cell DIVE mIF 平台(徕卡显微系统公司)采用碱性过氧化氢溶液作为荧光团灭活试剂,依次进行染色、成像和漂白,适用于 FFPE 切片。标准漂白工艺偶尔会出现漂白效率不理想、组织结构退化和抗原免疫原性丧失等问题。为了克服这些障碍,我们采用了一种改良的光化学漂白方法,即在使用过氧化氢的同时使用强 LED 光照射。在乳腺组织和其他组织切片上使用不同的抗体进行重复染色/漂白。比较了传统漂白和改良技术后的残留信号,并评估了组织完整性和抗原免疫原性。改良技术能有效消除前几轮染色产生的荧光信号,并在多轮染色和成像中产生一致的结果。采用改进的方法,光化学处理不会破坏组织亚细胞内容物,组织抗原性在整个 mIF 过程中得到了很好的保存。在 8 轮 mIF 过程中,整体处理时间从 36 小时缩短到 30 小时。而采用传统方法,8 轮处理后组织质量会严重下降。新技术缩短了周转时间,在 mIF 过程中可靠地去除荧光团,并很好地保持了组织的完整性,有助于研究多种生物标记物在相关组织中的共定位。
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引用次数: 0
Expression and Clinical Significance of Nuclear Phosphoglucomutase-1 in Hepatocellular Carcinoma. 核磷葡聚糖酶-1 在肝细胞癌中的表达和临床意义
IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2024-11-01 Epub Date: 2024-09-20 DOI: 10.1097/PAI.0000000000001225
Yechen Xia, Yan An, Riming Jin, Wentao Huang, Guang-Zhi Jin, Jing Xu

This study aimed to evaluate the predictive values of phosphoglucomutase-1 (PGM1) expression for prognosis in patients with hepatocellular carcinoma (HCC). PGM1 expression was assessed by immunohistochemistry in tissue microarrays. The relationship of PGM1 expression level with pathologic parameters and prognosis values was respectively analyzed by χ 2 test and Cox regression. The accuracy of independent risk factors in predicting prognosis was calculated by receiver operating characteristic curve. HCC patient-derived xenograft models were performed to evaluate the nuclear PGM1 antitumor effect. The results showed that PGM1 expression was low in HCC tissues. Nuclear PGM1 was an independent prognostic factor for overall survival and time to recurrence. Cox regression showed that nuclear PGM1, serum α-fetoprotein, liver cirrhosis, and TNM staging stage were independent risk predictors for HCC. Receiver operating characteristic curve demonstrated that combination of independent predictors had better prognostic value than TNM staging alone. Moreover, patient-derived xenograft models showed antitumor effect of nuclear PGM1. We found that low expression of nuclear PGM1 was detected in HCC tissues and associated with poor prognostic. Nuclear PGM1 was an independent prognostic factor in patients with HCC. Furthermore, nuclear PGM1 combining other independent risk factors showed a better prognostic value. Nuclear PGM1 was a useful prognostic biomarker for patients with HCC.

本研究旨在评估磷酸葡萄糖转氨酶-1(PGM1)表达对肝细胞癌(HCC)患者预后的预测价值。通过组织芯片中的免疫组织化学方法评估了 PGM1 的表达。通过χ2检验和Cox回归分别分析了PGM1表达水平与病理参数和预后值的关系。用接收者操作特征曲线计算了独立危险因素预测预后的准确性。对HCC患者来源的异种移植模型进行了核PGM1抗肿瘤作用的评估。结果显示,PGM1 在 HCC 组织中表达量较低。核 PGM1 是总生存期和复发时间的独立预后因素。Cox回归显示,核PGM1、血清α-胎儿蛋白、肝硬化和TNM分期是HCC的独立风险预测因子。接收者操作特征曲线显示,与单独的TNM分期相比,将独立的预测因子结合在一起具有更好的预后价值。此外,患者来源的异种移植模型显示了核 PGM1 的抗肿瘤作用。我们发现,核 PGM1 在 HCC 组织中的低表达与预后不良有关。核 PGM1 是 HCC 患者的一个独立预后因素。此外,核 PGM1 与其他独立危险因素相结合显示出更好的预后价值。核PGM1是一种对HCC患者有用的预后生物标志物。
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引用次数: 0
Comparing the Sensitivity of HER2 Epitope Detection of HercepTest mAb pharmDx (Dako Omnis, GE001) and Ventana PATHWAY Anti-HER-2/neu (4B5) Using IHC Calibrators. 使用 IHC 校准物比较 HercepTest mAb pharmDx(Dako Omnis,GE001)和 Ventana PATHWAY Anti-HER-2/neu (4B5) 检测 HER2 表位的灵敏度。
IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2024-11-01 Epub Date: 2024-10-24 DOI: 10.1097/PAI.0000000000001230
Frederik Aidt, Maria Sierra, Karin Salomon, Ghislain Noumsi

Accurate assessment of HER2 expression levels is paramount for determining eligibility for targeted therapies. HER2 immunohistochemistry provides a semiquantitative measurement of HER2 protein overexpression. Historically, little focus has been on the lower end of the HER2 expression range. The advent of novel therapeutic molecules that require fewer membrane epitopes to be effective has prompted a reevaluation of the current immunohistochemistry testing protocols, with special emphasis on the detection limit. Here, we have used Boston Cell Standards technology to determine the sensitivity of 2 commercially available HER2 immunohistochemistry assays, including a lower limit of detection.

准确评估 HER2 表达水平对于确定是否有资格接受靶向治疗至关重要。HER2 免疫组化可对 HER2 蛋白过表达进行半定量测量。一直以来,人们很少关注 HER2 表达范围的低端。新型治疗分子的出现减少了对膜表位的需求,促使人们重新评估当前的免疫组化检测方案,并特别强调检测限。在此,我们利用波士顿细胞标准技术确定了两种市售 HER2 免疫组织化学检测方法的灵敏度,包括检测下限。
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引用次数: 0
A Pilot Study on BRCA1/2 and PI3K Mutations Across Subtypes of Triple Negative Breast Cancer in North Indian Population. 北印度人群三阴性乳腺癌亚型中 BRCA1/2 和 PI3K 基因突变的试点研究
IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2024-11-01 Epub Date: 2024-11-07 DOI: 10.1097/PAI.0000000000001231
Parul Gupta, Tamanna Thakur, Anjali Chadda, Santosh Irinike, Siddhant Khare, Amanjit Bal

BRCA1/2 are tumor suppressor genes which regulate the DNA repair mechanism. Mutations in BRCA1/2 may increase the risk of breast cancer in patients. In the present study frequency of BRCA1/2 mutations in triple negative breast cancer (TNBC) patients was assessed and correlated with molecular subtypes of TNBC. Blood samples from 65 confirmed cases of TNBC were collected. DNA was isolated from whole blood and libraries were prepared using a BRCA1/2 custom panel. Sequencing was done on Ion torrent S5 sequencer and ion reporter was used for data analysis. Further molecular subtyping of mutation positive TNBC cases was done using immunohistochemistry markers CK5/6; CK4/14; Vimentin and E-Cadherin and androgen receptor (AR) using tissue microarray. Twenty five of 65 patients had heterozygous pathogenic mutations, alterations with conflicting interpretation of pathogenicity, variants of uncertain significance and variants of unknown significance. Nine patients had pathogenic mutation in BRCA 1 gene only and 2 patients had pathogenic mutations in BRCA2 gene. Two patients were transheterozygous for BRCA mutations, that is, had pathogenic mutations in both BRCA1/2 genes simultaneously and 5 were compound heterozygous (involving BRCA2 gene in all the cases). Prevalent subtypes among BRCA positive cases were unclassified subtype (n=4, 33%), Basal like (n=5, 41%), and mesenchymal subtype (n=3, 25%). None of the LAR subtype showed BRCA1/2 mutations. The present study observed that the BRCA1 mutation is more frequent than BRCA2 mutation in TNBC. BRCA1/2 mutations do not correspond to BRCAness or basal phenotype. Considering high incidence of breast cancer and lack of correlation of basal morphology with BRCA1/2 mutation, the molecular methods should be used for screening for BRCA1/2 mutations. This will not only help in familial screening but also in deciding targeted therapy with PARP (poly-ADP ribose polymerase) inhibitors.

BRCA1/2 是调节 DNA 修复机制的肿瘤抑制基因。BRCA1/2 基因突变可能会增加患者罹患乳腺癌的风险。本研究评估了三阴性乳腺癌(TNBC)患者中 BRCA1/2 基因突变的频率,并将其与 TNBC 的分子亚型相关联。研究收集了 65 例 TNBC 确诊病例的血液样本。从全血中分离出 DNA,并使用 BRCA1/2 定制面板制备文库。测序在 Ion torrent S5 测序仪上进行,数据分析使用离子报告器。利用免疫组化标记物CK5/6、CK4/14、波形蛋白和E-Cadherin以及组织芯片中的雄激素受体(AR)对突变阳性的TNBC病例进行了进一步的分子亚型鉴定。65 例患者中有 25 例存在杂合致病突变、致病性解释不一致的变异、意义不确定的变异和意义不明的变异。9 名患者仅有 BRCA 1 基因的致病突变,2 名患者有 BRCA2 基因的致病突变。两名患者是 BRCA 基因突变的跨杂合型,即 BRCA1/2 基因同时发生致病性突变,5 名患者是复合杂合型(所有病例都涉及 BRCA2 基因)。BRCA 阳性病例的常见亚型为未分类亚型(4 例,33%)、基底样亚型(5 例,41%)和间质亚型(3 例,25%)。LAR亚型均未发现BRCA1/2基因突变。本研究发现,在 TNBC 中,BRCA1 突变比 BRCA2 突变更常见。BRCA1/2突变与BRCAness或基础表型并不对应。考虑到乳腺癌的高发病率以及基底形态与 BRCA1/2 基因突变缺乏相关性,应采用分子方法筛查 BRCA1/2 基因突变。这不仅有助于家族性筛查,还有助于决定使用 PARP(聚-ADP 核糖聚合酶)抑制剂进行靶向治疗。
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引用次数: 0
Automated Standardization of Melanin Bleaching Procedures of Heavily Pigmented Melanocytic Lesions With Low-Concentration Hydrogen Peroxide on an Automated Platform. 在自动化平台上使用低浓度过氧化氢对严重色素沉着的黑色素细胞病变进行黑色素漂白程序的自动化标准化。
IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2024-10-01 Epub Date: 2024-08-12 DOI: 10.1097/PAI.0000000000001217
Chih-Ching Yeh, Yi-Jing Li, Jang-Shian Liang, Jia-Bin Liao

Melanin is a natural pigment in the human body that is primarily found in the skin and hair. It protects the skin from damage by ultraviolet radiation. Although this pigment plays a crucial role in protecting the human body, it represents a challenge for pathologists to evaluate highly pigmented tissue samples from melanoma or pigmented skin lesions. Abundant melanin may obscure tissue morphology, which makes it very difficult for pathologists to make a differential diagnosis. Melanin pigment is brown-to-black and granular, and its distribution is often uneven in tissues. The presence of these pigments can complicate the analysis of immunohistochemistry (IHC) for 2 reasons. First, they have a direct physical masking effect on antigen-antibody interactions. Second, 3,3-diaminobenzidine, the most commonly used chromogen, has a brown color that is difficult to distinguish from melanin pigment. Therefore, melanin bleaching has become a crucial step in handling pigmented melanocytic lesions. Bleaching techniques aid pathologists in histopathologic examination of melanin-rich tissue. In this study, we integrated melanin bleaching and IHC on an automated IHC platform to set up a rapid and fully automated procedure. Bleaching steps were performed before antigen retrieval. Samples were treated with 1% hydrogen peroxide solution in Tris-HCl buffer (pH 10) at 80°C for 8 minutes, achieving optimal conditions for melanin bleaching while preserving tissue morphology and antigenicity. This rapid, effective, fully automated, and standardized workflow can be applied to routine staining procedures in clinical laboratories, thereby improving the quality of pathological diagnosis.

黑色素是人体内的一种天然色素,主要存在于皮肤和头发中。它能保护皮肤免受紫外线辐射的伤害。虽然这种色素在保护人体方面起着至关重要的作用,但病理学家在评估黑色素瘤或色素性皮肤病变的高色素组织样本时却面临着挑战。丰富的黑色素可能会遮盖组织形态,这使得病理学家很难做出鉴别诊断。黑色素色素呈棕黑色颗粒状,在组织中的分布往往不均匀。这些色素的存在会使免疫组化(IHC)分析复杂化,原因有二。首先,它们对抗原-抗体的相互作用有直接的物理遮蔽作用。其次,最常用的显色剂 3,3-二氨基联苯胺呈棕色,很难与黑色素区分开来。因此,黑色素漂白已成为处理色素性黑素细胞病变的关键步骤。漂白技术有助于病理学家对富含黑色素的组织进行组织病理学检查。在这项研究中,我们在一个自动 IHC 平台上整合了黑色素漂白和 IHC,建立了一个快速、全自动的程序。漂白步骤在抗原检索之前进行。样本在 80°C 下用 Tris-HCl 缓冲溶液(pH 10)中的 1% 过氧化氢溶液处理 8 分钟,以达到黑色素漂白的最佳条件,同时保留组织形态和抗原性。这种快速、有效、全自动和标准化的工作流程可应用于临床实验室的常规染色程序,从而提高病理诊断的质量。
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Applied Immunohistochemistry & Molecular Morphology
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