乳腺导管内增生病变中p53表达与外显子突变的研究

目的 肿瘤抑制基因p53异常是浸润性乳腺癌发生发展中常见事件,而其与包括普通型增生(usual ductal hyperplasia,UDH)、不典型增生 (atypical ductal hyperplasia,ADH)及导管内原位癌 (ductal carcinoma in situ,DCIS)的乳腺导管内增生性病变关系不明.本研究旨在探讨乳腺导管内增生性病变中p53外显子突变及突变型p53蛋白表达情况,以期了解p53突变及蛋白表达在乳腺癌发生发展中的作用.方法 用高分辨率熔解曲线(high-resolution melting,HRM)结合测序研究140例乳腺导管内增生性病变中p53外显子5-8的突变情况.用免疫组化研究240例乳腺导管内增生性病变中突变型p53蛋白表达情况.结果 经过HRM分析,共17例患者DNA熔解曲线与野生型标准品熔解曲线大于阈值结合测序分析结果发现,其中16例出现p53外显子突变.p53在UDH、ADH及DCIS中的突变率为0.0%(0/40),12.7%(8/63)和21.6%(8/37),三者间差异显著(P<0.05).40例UDH中未出现突变型p53蛋白阳性表达,在14.6%(19/130)的ADH出现阳性表达,在31.4%(22/70)的DCIS中出现阳性表达,三者间差异显著(P<0.05).Spearman相关性分析示突变型p53蛋白表达与p53外显子突变呈正相关(r=0.792,P<0.01).结论 p53外显子突变及突变型p53蛋白表达发生于乳腺导管内增生性病变中的ADH与DCIS,其可能为乳腺癌发生发展中的早期事件.     

乳腺导管内增生性病变的分类及其临床病理特点

乳腺导管内增生性病变不仅是病理诊断的难点,也是乳腺外科治疗方案选择不统一的问题所在。2003年WHO乳腺肿瘤组织学分类将其分为普通型导管增生(usual ductalhyperplasia,UDH)、平坦型上皮不典型增生(flat epithelial atypia,FEA)、不典型导管增生(atypical ductal hyperplasia,ADH)和导管原位癌(ductal carcinoma in situ,DCIS)4类。UDH的细胞学及组织学构型均无不典型性,ADH仅有细胞学不典型性而无组织构型不典型性,DCIS则具有细胞学和组织构型的双重不典型性,而FEA中的少数应视为癌前病变。微小钙化是导管内增生性病变常见的X线表现形式。乳腺摄影能早期发现临床触及不到的微小癌及早期癌,尤其是DCIS,有助于临床手术范围或手术方式的选择。     

SOX10在乳腺导管上皮性病变病理诊断及鉴别诊断中的作用

目的:探讨SOX10在乳腺导管上皮普通性增生(usual ductal hyperplasia, UDH)、非典型增生(atypical hyperplasia, ADH)、导管内癌(ductal carcinoma in situ, DCIS)及不同亚型浸润性导管癌中的表达情况。方法:收集我院病理科2021年12月至2023年06月进行了SOX10免疫组化染色的乳腺标本共197例,其中UDH 48例、ADH 20例、DCIS 40例、伴大汗腺分化的癌10例、luminal A 20例、luminal B 27例、Her2过表达10例及三阴性乳腺癌(triple-negative breast carcinoma, TNBC)22例,分析不同乳腺导管上皮性病变中SOX10的表达差异。结果:SOX10在48例UDH呈马赛克样斑驳着色,而20例ADH及40例DCIS中均不表达,差异具有统计学意义(P<0.001);SOX10在三阴性乳腺癌中阳性率为68.20%(15/22),而在luminal A型、luminal B型、Her2过表达型浸润性乳腺癌中均不表达,差异具有统计学意义(P...     

乳腺导管内增生病变中p53表达与外显子突变的研究

目的：肿瘤抑制基因p53异常是浸润性乳腺癌发生发展中常见事件,而其与包括普通型增生（usualductal hyperplasia,UDH）、不典型增生（atypical ductal hyperplasia,ADH）及导管内原位癌（ductal carcinoma insitu,DCIS）的乳腺导管内增生性病变关系不明。本研究旨在探讨乳腺导管内增生性病变中p53外显子突变及突变型p53蛋白表达情况,以期了解p53突变及蛋白表达在乳腺癌发生发展中的作用。方法：用高分辨率熔解曲线（high-resolution melting,HRM）结合测序研究140例乳腺导管内增生性病变中p53外显子5-8的突变情况。用免疫组化研究240例乳腺导管内增生性病变中突变型p53蛋白表达情况。结果：经过HRM分析,共17例患者DNA熔解曲线与野生型标准品熔解曲线大于阈值结合测序分析结果发现,其中16例出现p53外显子突变。p53在UDH、ADH及DCIS中的突变率为0.0%（0/40）,12.7%（8/63）和21.6%（8/37）,三者间差异显著（P〈0.05）。40例UDH中未出现突变型p53蛋白阳性表达,在14.6%（19/130）的ADH出现阳性表达,在31.4%（22/70）的DCIS中出现阳性表达,三者间差异显著（P〈0.05）。Spearman相关性分析示突变型p53蛋白表达与p53外显子突变呈正相关（r=0.792,P〈0.01）。结论：p53外显子突变及突变型p53蛋白表达发生于乳腺导管内增生性病变中的ADH与DCIS,其可能为乳腺癌发生发展中的早期事件。     

乳腺癌旁导管内增生性病变中ER、Ki-67的表达及意义

[目的]探讨ER、Ki-67在乳腺癌旁导管内增生性病变中的表达及意义。[方法]采用免疫组化EnVision法对90例乳腺浸润性导管癌（invasive duetal carcinoma，IDC），癌旁乳腺导管内增生性病变（intraductal proliferative lesion，IDPL）进行ER、Ki-67、CK5／6、CK34βE12、p63、SMA染色标记。[结果]ER在正常终末导管小叶单位（terminal duct-lobular unit，TDLU），普通型导管增生（usual ductal hyperplasia，UDH），非典型性导管增生（atypical ductal hyperplasia，ADH），原位导管癌（ductal carcinoma in situ，DCIS）中的阳性细胞数逐渐增加，并从由少量散在到不连续、连续的片状分布逐渐转变，其中ADH组明显高于UDH组（P〈0．01），DCIS组显著高于ADH组（P〈0．05）。Ki-67在平坦型上皮不典型性（flat epithelial atypia，FEA），DCIS中的表达明显高于ADH（P〈0．05），Ki-67的表达在UDH、ADH中无显著性差异。[结论]ER在UDH、ADH、DCIS中的表达逐渐增加，且两两间均有显著性差异，ER的过表达可能是IDPL恶性转化过程中的改变之一：Ki-67的表达在DCIS组中明显高于ADH组中，Ki-67的表达可能是某些乳腺癌发生的早期事件，两者可作为评估IDPL发生IDC的危险性的生物学指标。     

34βE12、CK5/6、p63、SMA在乳腺增生、不典型增生与原位癌鉴别诊断中的价值

目的探讨高分子量细胞角蛋白34βE12、细胞角蛋白CK5/6、肌上皮标记物p63、平滑肌肌动蛋白SMA在乳腺增生、不典型增生与原位癌鉴别诊断中的价值。方法采用免疫组织化学染色法(SP),检测34βE12、CK5/6、p63、SMA在30例乳腺普通型增生(UDH)、30例非典型导管增生(ADH)、20例导管原位癌(DCIS)和20例浸润性导管癌(IDC)中的表达。结果 UDH和ADH组中34βE12均呈阳性表达,DCIS和IDC组中大部分34βE12呈阴性表达,前两组与后两组比较,差异均有统计学意义(P<0.05);UDH组中CK5/6均呈阳性表达,ADH、DCIS及IDC组中CK5/6大部分呈阴性表达,UDH组与其他3组比较差异有显著性(P<0.05);p63表达量依次为UDH组>ADH组>DCIS组,IDC组中其无阳性表达,各组间差异均有统计学意义(P均<0.05);UDH、ADH和DCIS组中SMA均呈阳性表达,IDC组中其大部分呈阴性表达,与前3组比较,差异均有统计学意义(P均<0.05)。结论联合检测34βE12、CK5/6、p63、SMA表达,有助于鉴别乳腺增生、不典型增生与原位癌,为临床提供可靠的诊疗依据。     

维吾尔族乳腺癌患者JAG1基因甲基化的临床意义北大核心CSCD

目的探讨JAG1甲基化在维吾尔族乳腺癌变和进展过程中的临床意义。方法 MALDI-TOF MS法检测JAG1基因在15例乳腺普通型导管增生(usual ductal hyperplasia,UDH)、15例非典型性导管增生(atypical ductal hyperplasia,ADH)、15例导管原位癌(ductal carcinoma in situ,DCIS)和50例浸润性导管癌(invasive ductal carcinoma,IDC)组织中的甲基化率,免疫组织化学法检测蛋白表达,分析甲基化与蛋白表达的相关性。结果 JAG1基因在乳腺癌变过程组织中的甲基化率逐渐降低,以Cp G_11.12、Cp G_14.15、Cp G_18.19最显著(P<0.05),Cp G_1在低分化组明显降低(P<0.05),Cp G_8.9.10在淋巴结转移组明显降低(P<0.05),而Cp G_16.17在Ⅲ期中显著升高(P<0.05),Cp G_13、Cp G_26在ER、PR和HER2表达间差异有统计学意义(P<0.05),JAG1基因甲基化与蛋白表达显著负相关(r=-0.6 7 4 P<0.001)。结论JAG1基因CpG位点的不同甲基化水平在乳腺癌变进展中可能发挥不同作用。JAG1甲基化是蛋白表达的调控机制之一。     

Jagged1基因甲基化与乳腺癌发生发展相关性探讨

目的 探讨Jagged1基因异常甲基化在乳腺癌发生发展过程中的意义.方法 采用MALDI-TOF MS法检测2004-01-16-2009-06-25石河子大学第一附属医院63例乳腺浸润性导管癌(invasive ductal carcinoma,IDC)、20例导管原位癌(ductal carcinoma in situ,DCIS)、20例非典型导管增生(atypical ductal hyperplasia,ADH)和20例普通型导管增生(usual ductal hyperplasia,UDH)组织中Jagged1基因甲基化水平,免疫组织化学方法检测4组乳腺组织中Jagged1蛋白表达状况,在IDC组中进一步分析Jagged1甲基化和表达与临床病理特征的相关性.结果 Jagged1基因总甲基化率在IDC(0.127 6±0.067 5)、DCIS(0.138 9±0.093 1)、ADH(0.168 0±0.014 6)和UDH(0.223 3±0.060 9)中逐渐升高,并且IDC组甲基化率分别与ADH(P=0.015)和UDH(P＜0.001)组比较,差异有统计学意义.CpG-2、CpG-6、CpG-13、CpG-20.21.22、CpG-23.24.25、CpG-26位点的甲基化率在IDC组最低,差异有统计学意义,P值均＜0.05.Jagged1蛋白在IDC(58.7％,37/63)、DCIS(45.0％,9/20)、ADH(40.0％,8/20)和UDH(25.0％,4/20)组中阳性表达率逐渐降低,其中IDC组的阳性率显著高于UDH组,P=0.003.Jagged1蛋白高表达与DNA低甲基化在IDC(P＜0.001)、DCIS(P=0.003)、ADH(P=0.004)和UDH组(P=0.007)均有相关性.Jagged1基因低甲基化和蛋白高表达与临床分期(P＜0.001;P=0.019)和组织学分级(P=0.003;P=0.025)均有相关性.结论 Jagged1基因低甲基化和CpG位点低甲基化,可能参与乳腺癌的发生,并且Jagged1基因低甲基化可能是调控蛋白高表达的方式之一,进而促进乳腺的癌变和进展.     

伴和不伴癌的乳腺导管内增生性病变ERα与p53蛋白表达差异的研究

目的:研究伴癌的乳腺导管内增生性病变与不伴癌的乳腺导管内增生性病变ERα与p53蛋白表达的差异。方法:用免疫组化法研究129例不伴癌的乳腺导管内增生性病变及86例伴癌的乳腺导管内增生性病变中p53及ERα蛋白表达差异。结果:p53蛋白阳性表达于18.2%(14/77)单纯性非典型导管增生(ADH),31.0%(9/29)伴原位癌的ADH(ADH/DCIS),26.7%(8/30)伴浸润性导管癌的ADH(ADH/IDC),三者间表达率差异无统计学意义,P>0.05。ERα在79.3%(23/29)伴原位癌的ADH(ADH/DCIS)中阳性表达,在76.7%(23/30)的伴浸润性导管癌的ADH(ADH/IDC)中阳性表达,均明显低于其在单纯性ADH中的表达(72/77,93.5%),P<0.05。在乳腺导管内增生性病变中,突变型p53蛋白的核聚集与ERα阳性表达的相关系数r=-0.512,P<0.01。结论:初步研究结果提示,虽然不伴癌的ADH与伴癌的ADH有相同的形态学表现,但形成的分子机制可能存在差异。     

应用组织芯片技术研究p27kip1和Skp2在星形胶质细胞瘤及增生中的表达和意义

目的:探讨p27kip1和Skp2在星形胶质细胞增生及星形胶质细胞瘤中的表达及其与肿瘤发生发展的关系。方法:利用组织芯片技术及PV6000通用型二步法免疫组化方法检测p27kip1和Skp2在正常脑组织,星形胶质细胞增生,低和高级别星形胶质细胞瘤中的表达。结果:正常脑组织中p27kip1阳性表达率91.7%,Skp2表达阴性;增生组中p27kip1和Skp2阳性表达率分别为86.4%、28.6%,与正常组比较,p27kip1无统计学意义;Skp2有统计学意义;低级别肿瘤组中二者阳性表达率分别为64.3%、46.7%,与增生组比较,p27kip1有统计学意义;Skp2无统计学意义;高级别肿瘤组中二者阳性表达率分别为42.6%、69.6%,与低级别肿瘤组比较,差异均有统计学意义;p27kip1和Skp2与组织学分级密切相关。结论:p27kip1有可能成为鉴别星形胶质细胞增生与低级别星形胶质细胞瘤的客观指标,且p27kip1和Skp2在星形胶质细胞瘤的发生发展过程中有重要作用。     

p53 nuclear accumulation and ERα expression in ductal hyperplasia of breast in a cohort of 215 Chinese women

Introduction

Women with ductal hyperplasia including usual ductal hyperplasia (UDH) and atypical ductal hyperplasia (ADH) have an increased risk of developing invasive ductal carcinoma (IDC) of breast. The importance of several molecular markers in breast cancer has been of considerable interest during recent years such as p53 and estrogen receptor alpha (ERα). However, p53 nuclear accumulation and ERα expression have not been assessed in ductal hyperplasia co-existing with ductal carcinoma in situ (DCIS) or IDC versus pure ductal hyperplasia without DCIS or IDC.

Materials and methods

We investigated p53 nuclear accumulation and ERα expression in breast ductal hyperplasia in a cohort of 215 Chinese women by immunohistochemistry (IHC), which included 129 cases of pure ductal hyperplasia, 86 cases of ductal hyperplasia co-existing with DCIS (41 cases) or IDC (45 cases).

Results

Nuclear p53 accumulation was identified in 22.8% of ADH (31/136), 41.5% of DCIS (17/41) and 42.2% of IDC (19/45), and no case of UDH (0/79). No difference in nuclear p53 accumulation was observed between pure ADH and ADH co-existing with DCIS (ADH/DCIS) or IDC (ADH/IDC) (P > 0.05). The positive rate of ERα expression was lower in ADH (118/136, 86.8%) than that in UDH (79/79, 100%) (P < 0.001), but higher than that in DCIS (28/41, 68.3%) or IDC (26/45, 57.8%) respectively (P < 0.001). The frequency of ERα expression was lower in ADH/DCIS (23/29, 79.31%) and ADH/IDC (23/30, 76.67%) than that in pure ADH (72/77, 93.51%) respectively (P < 0.05). There was a negative weak correlation between p53 nuclear accumulation and ERα expression as for ADH (coefficient correlation -0.51; P < 0.001).

Conclusions

Different pathological types of ductal hyperplasia of breast are accompanied by diversity in patterns of nuclear p53 accumulation and ERα expression. At least some pure ADH is molecularly distinct from ADH/CIS or ADH/IDC which indicated the two types of ADH are molecularly distinct entities although they have the same morphological appearance.     

Expression of ER, Ki-67 and CylinD1 in the Pre-cancerous Breast of Chinese Patients

To investigate the expression and association of ER, Ki-67 and cyclinD1 in usual ductal hyperplasia(UDH), atypical ductal hyperplasia (ADH) and ductal carcinoma in situ(DCIS) in the breast. The study included 56 cases of pre-cancerous lesions which were surgically excised at Qi Lu Hospital of Shangdong University. Immunohistochemistry was used to determine the expression of ER, Ki-67 and cyclinD1 and double-labelling immunofluorescence technique was used to observe the coexpression of ER and Ki-67. The expression and distribution of ER-positive cells were significantly different in UDH, ADH and DCIS. The ER-positive cells were much more in UDH than in normal TDLUs (terminal duct lobular units). The distribution of ER-positive cells interspersed amid ER-negative cells within UDH. However , the ER positive cells showed marked increases in ADH and low grade nuclear DCIS (P < 0.05), distributing in almost all constituent cells. The expression of ki-67 and cyclinD1 were significantly different between UDH and DCIS (P < 0.05) , and a positive correlation was found between expression of Ki-67 and morphological classification of pre-cancerous lesions (r = 0.3522, P < 0.05) as well as cyclinD1 (r = 0.3901, P < 0.05). Double-labelling immunofluorescence showed that there was no coexpression of ER and Ki-67 in normal breast tissue. The coexpression of the two markers was found in ADH and increased in DCIS. Overexpression of ER, Ki-67 and cyclinD1 significantly accompanies the transition of normal cells and UDH to ADH and DCIS. The coexpression of ER and ki-67 may present the early change in carcinogenesis of breast cancer.     

Evidence of chromosomal alterations in pure usual ductal hyperplasia as a breast carcinoma precursor

Previous studies have shown the chromosomal alterations in usual ductal hyperplasia (UDH), atypical ductal hyperplasia (ADH), and ductal carcinoma in situ (DCIS) in the breast with bilateral ductal hyperplasia or adjacent to invasive ductal carcinoma (IDC). However, the role of UDH as a putative precursor of breast IDC is not clear and has not been fully addressed. The aim of this study was to clarify the role of UDH in breast carcinoma pathogenesis. To investigate chromosomal imbalances and commonality, samples of pure unilateral UDH (n=20) were obtained by laser capture microdissection and analyzed by comparative genomic hybridization. Other ductal lesions, including ADH (n=2), high-grade DCIS (n=3), and grade III IDC (n=5), were assessed at the same time for comparison. The mean values of alteration were 1.95 (39/20) in UDH, 9.5 (19/2) in ADH, 11.0 (33/3) in DCIS and 18.2 (89/5) in IDC, respectively. Some common predisposition regions for the deletions were at chromosomes 1p36-pter, 13q11-14, and 16q11-23, while the high frequency amplification regions were 1q31-qter, 3p21-pter, 6p21-pter, 11q11-14, 12q11-qter, 13q21-qter, 16p12-pter, 17q12-22, and 20q. The genetic abnormalities in the spectrum of breast ductal hyperplasia revealed that the deletion of DNA copy in UDH was the lowest, and gradually increased in the lineages of ADH, DCIS and IDC. Results showed that a significant portion of UDH shares common genetic alterations with ADH, DCIS and IDC, indicating UDH as a precursor of invasive breast ductal carcinoma.     

Preliminary Study on the Expression and the Significance of S-Phase Kinase Associated Protein 2 in Atypical Ductal Hyperplasia and Breast Cancer

In 1995 Skp2 was cloned by Zhang et al.[1]from human fibroblasts and it was shown that Skp2 interacts by combining with cyclin de- pendent kinase 2 (cyclin-CDK2). Skp2 is a member of the F-box fam- ily of substrate-recognition subunits of Skp2-Cullin-F-bo…     

Analysis of the progression of intraductal proliferative lesions in the breast by PCR-based clonal assay

Purpose To analyze the progression in patients with a morphological diagnosis of intraductal proliferative lesions by PCR-based clonal assay. Materials and methods An X-chromosome inactivation assay was applied to explore clonal relationships in human intraductal proliferative lesions of the breast. Four groups samples, including 40 cases of usual ductal hyperplasia (UDH), 40 cases of atypical ductal hyperplasia (ADH), 29 cases of flat epithelia atypia (FEA), and 40 cases of ductal carcinoma in situ (DCIS) were selected for analysis. Thirty specimens of normal breast tissue were used as a control group. Microdissection was performed to collect the tissue samples for extraction of genomic DNA from paraffin-embedded tissues. The DNA was subjected to PCR amplification of the CAG repeats in androgen receptor (AR) gene exon I with and without prior digestion of methylation-sensitive restriction enzyme HhaI. Gel electrophoresis was used to detect the clonal nature of these four groups samples. Results The clonal analysis confirmed monoclonality in all informative samples of DCIS cells. Normal tissues and the majority (97.1%) of UDH were shown to be polyclonal. Monoclonality was revealed in 20/39 (51.3%) cases of ADH. Among 26 cases of FEA, 20 were shown to be polyclonal, while six displayed monoclonal alterations which accounted for 23.1%. Conclusion These findings reinforce recent suggestions that clonal analysis with AR gene polymerase chain reaction may be used to define the genetic relationships among the human tumor and the breast intraductal proliferative lesions. Furthermore, our observations demonstrate nearly a half ADH and the smaller part of FEA have clonal alterations, which may be neoplastic lesions. This method would shed light on genetic abnormalities that play a role in early tumorigenesis of the breast, and thus might be an adjunct in predicting the probability of breast tumor occurrence and in guiding the management of these cases.     

Skp2和P27kip1在前列腺癌组织中的表达与临床病理特征的相关性研究

背景及目的:F-box蛋白Skp2参与细胞周期抑制蛋白P27kip1泛素化降解,研究发现Skp2在乳腺癌、胃癌、前列腺癌等多种恶性肿瘤中表达增加.本研究旨在探讨Skp2和P27kip1在前列腺癌组织中的表达及与前列腺癌各项临床病理特征的关系,并探讨两者的相关性.方法:用免疫组化EnVisionTM方法检测Skp2和P27 kip1蛋白在41例前列腺癌和20例良性前列腺增生组织中的表达情况.结果:在前列腺癌中的Skp2蛋白阳性率[(8.52±2.40)%]显著高于良性前列腺增生[(0.21±0.15)%](P＜0.001).Skp2蛋白表达与前列腺癌术前血清前列腺特异性抗原(PSA)水平(r=0.360,P=0.021)、局部浸润(r=0.570,P＜0.001)、肿瘤分期(r=0.531,P＜0.001)、病理分级(r=0.514,P=0.001)呈正相关.在前列腺癌中的P27kip1蛋白阳性率[(70.71±4.25)%]显著低于良性前列腺增生[(97.21±2.10)%](P＜0.001).P27kip1蛋白表达与前列腺癌术前PSA水平(r=-0.399,P=0.010)、局部浸润(r=-0.329,P=0.036)、肿瘤分期(r=-0.453,P=0.003)、病理分级(r=-0.290,P=0.046)呈负相关.前列腺癌中P27kip1蛋白与Skp2表达呈负相关(r=-0.572,P＜0.001).结论:前列腺癌中Skp2蛋白表达与靶蛋白P27 kip1蛋白降解有关,提示Skp2蛋白可能与前列腺癌的发生发展有关.     

乳腺癌和乳腺导管内增生性病变JAG1基因甲基化的检测及临床意义

目的:探讨JAG1基因甲基化在乳腺浸润性导管癌发病中的作用。方法:采用MassARRAY方法对乳腺浸润性导管癌(IDC;n=75)、乳腺导管原位癌(DCIS;n=23)、乳腺非典型性导管增生症(ADH;n=20)以及乳腺普通型导管增生症(UDH;n=27)进行JAG1基因甲基化的定量检测。结果:JAG1基因启动区CpG_13、CpG_20.21.22、CpG_26位点在UDH组中的平均甲基化率高于ADH、DCIS、IDC组（P<0.05）。结论:在乳腺癌患者中JAG1(该基因的表达与乳腺癌细胞的生长、浸润、转移、预后有关)基因的部分位点呈现低甲基化改变。CpG_13、CpG_20.21.22、CpG_26位点的低甲基化改变与乳腺癌的形成具有相关性,可能该基因特异性CpG位点的低甲基化参与了乳腺良性病变向乳腺癌的发展演进过程。