【作者】 齐立强；

【导师】 付丽；

【作者基本信息】 天津医科大学 ， 肿瘤学， 2009， 博士

【摘要】 [背景]乳腺平坦型上皮不典型性(flat epithelial atypia, FEA)的特征是一到几层的轻度不典型管腔上皮细胞。导管内原位癌(dutal carcinoma in situ, DCIS)和浸润性导管癌非特殊型(invasive dutal carcinoma, not otherwise specified, IDC-NOS)的遗传学变化也可见于FEA,尽管发生的几率不一样。至今,FEA中miRNA的表达及其与DCIS和IDC的关联,尚未见报道。[方法](1)从270例DCIS中选出51例含FEA的病例,根据形态学表现将其分为4级,Ⅰ、Ⅱ级为低分级,Ⅲ、Ⅳ级为高分级；根据与DCIS的位置关系将其分为两组,并进行ER, PR,α-SMA, p63, p53, bcl-2, Her-2, cyclinD1, Ki-67免疫组化染色。(2)对15例同时存在正常乳腺,FEA和/或DCIS以及17例IDC-NOS病例行miRNA-21原位杂交(in situ hybridization, ISH)。同时,用免疫组化(immunohistochemistry, IHC)方法研究miR-21的靶基因PDCD4,TM1和PTEN的表达。[结果](1)Ⅰ～Ⅳ级FEA分别为28、8、6和9例；与DCIS位于同一象限的24例,与DCIS位于不同象限的27例,且随着FEA分级从低到高,其与DCIS位于同一象限的比例逐渐增加(Ⅰ、Ⅱ、Ⅲ、Ⅳ级分别为28.6%、37.5%、66.7%、100%)。免疫组化结果：α-SMA和p63染色显示,FEA的肌上皮细胞明显受压变薄。其余各项免疫指标的阳性表达率分别为ER (62.8%), PR (54.9%), p53 (21.6%), bcl-2 (62.8%), Her-2 (15.7%), cyclinD1 (62.8%), Ki-67 (35.3%).低分级FEA、高分级FEA和DCIS三者比较,Her-2、cyclinD1和Ki-67的表达存在差异,高分级FEA和DCIS的Her-2、cyclinD1、Ki-67阳性率均明显高于低分级FEA,但高分级FEA和DCIS间无明显差别；在位置关系上,与DCIS位于同一象限的FEA、与DCIS位于不同象限的FEA和DCIS者比较,Her-2、cyclinD1和Ki-67的表达也存在差异,且与DCIS位于不同象限FEA的Her-2、cyclinD1、Ki-67阳性率均明显低于DCIS,但与DCIS位于同一象限FEA和DCIS间无明显差别。(2)15例正常乳腺当中有2例,15例FEA当中有7例,12例DCIS当中有9例,17例IDC-NOS中有15例niR-21呈阳性表达。在12例同时存在正常乳腺,FEA及DCIS的病例中,我们发现有5例miR-21从正常乳腺到FEA进而到DCIS呈表达上升趋势。有3例FEA为阴性而DCIS为阳性；另外的3例FEA和DCIS均为miR-21阴性表达；最后一例全部三种成分(正常乳腺,FEA和DCIS) miR-21皆为阳性表达。总之,我们观察到miR-21的阳性表达率从正常乳腺到FEA,DCIS进而到IDC-NOS有逐步增加的趋势。IHC结果表示：PTEN表达强度在四种成分(正常乳腺,FEA, DCIS和IDC-NOS)中没有明显差异；胞浆中的PDCD4染色从正常乳腺到IDC-NOS逐步升高,而胞核中的PDCD4染色从正常乳腺到IDC-NOS却逐步下降。TM1染色从正常乳腺中的阳性表达下降到在多数DCIS和IDC-NOS中呈阴性表达。而在FEA中,TM1的染色表型和正常乳腺相似。【结论】(1)FEA分级越高,与DCIS的位置关系越密切。FEA(特别是高分级的、与DCIS位置关系密切的FEA)作为一种具有恶性潜能的交界性病变,具有一定的意义。(2) miR-21从乳腺正常导管上皮到FEA, DCIS和IDC-NOS的上调表达与肿瘤发生的形态学演变呈平行变化趋势。我们没有观察到miR-21和先前报道的它的靶向基因之间明确的关系。【背景】与散发性乳腺癌和BRCA2相关的乳腺癌相比,BRCA1相关的乳腺癌有其独特的临床病理特征,DNA拷贝数的变化和基因表型特征。microRNA (miRNA)在一系列的生理病理学功能主面发挥重要作用,例如生长发育,增殖凋亡以及肿瘤形成等等。我们的研究目的是探索BRCA1相关的乳腺癌是否有其独特的miRNA表达谱特征。【方法】用miRNA microarray方法研究BRCA1相关和非相关乳腺癌的miRNA表型特征。本研究包括8例BRCA1相关乳腺癌,10例BRCA1/2非相关的散发性乳腺癌和6例正常乳腺(最后汇集成2个标本),比较不同类别标本的328个flagged miRNA表达的各自特点。【结果】鉴定出在BRCA1突变和非突变的乳腺癌中明显差异表达的miRNA亚群。并且,从中进一步选出一些可能在BRCA1通路中发挥一定作用的niRNA。【结论】层序聚类分析方法证实与BRCA1/2非突变的散发性乳腺癌相比较,BRCA1突变的乳腺癌有其独特的miRNA表达谱特征。更多还原

【Abstract】 [Background]Flat epithelial atypia (FEA) of the breast is characterised by a few layers of mildly atypical luminal epithelial cells. Genetic changes found in ductal carcinoma in situ (DCIS) and invasive ductal breast carcinoma, not otherwise specified (IDC-NOS) are also found in FEA, albeit at a lower concentration. So far, miRNA expression changes associated with invasive breast cancer, like miR-21, have not been studied in FEA.[Methods](1)51 cases of FEA were selected from 270 cases of DCIS with FEA components. The cases were divided into 4 grades and 2 groups, and immunohistochemical study for ER, PR,α-SMA, p63, p53, bcl-2, Her-2, cyclinD1 and Ki-67 was performed on these cases.(2) We performed miRNA in-situ hybridization (ISH) on 15 cases with simultaneous presence of normal breast tissue, FEA and/or DCIS and 17 additional cases with IDC-NOS. Expression of the miR-21 targets PDCD4, TM1 and PTEN was investigated by immunohistochemistry.[Results](1) The cases of gradeⅠ-Ⅳof FEA were 28,8,6 and 9, respectively. There were 24 cases of FEA located in the same quadrant with DCIS and 27 cases in the different quadrant. The proportion of FEA in the same quadrant with DCIS increased with the grade of FEA (gradeⅠ,Ⅱ,Ⅲ,Ⅳwere 28.6%,37.5%,66.7%,100%, respectively). Immunohistochemically, myoepithelial cells of FEA appeared attenuated stained with a-SMA and p63. The positive rate of FEA for ER, PR, p53, bcl-2, Her-2, cyclinD1 and Ki-67 were 62.8%,54.9%,21.6%,62.8%,15.7%,62.8% and 35.3%, respectively. There were significant difference in the expression of Her-2, cyclinDl and Ki-67 among low grade FEA, high grade FEA and DICS, and the positive rate of Her-2, cyclinD1 and Ki-67 were significantly higher in high grade FEA and DICS than in low grade FEA. There were significant difference in the expression of Her-2, cyclinD1 and Ki-67 among FEA in the same quadrant with DCIS, FEA in the different quadrant with DCIS and DICS, and the positive rate of Her-2, cyclinD1 and Ki-67 were significantly lower in FEA in the different quadrant with DCIS than in DCIS.(2) Two out of fifteen cases showed positive staining for miR-21 in normal breast ductal epithelium, seven out of fifteen cases were positive in the FEA component and nine out of twelve cases were positive in the DCIS component. A positive staining of miR-21 was observed in 15 of 17 IDC-NOS cases. In 12 cases all three components were present in one tissue block and an increase of miR-21 from normal breast to FEA and to DCIS was observed in 5 cases. In three cases the FEA component was negative, whereas the DCIS component was positive for miR-21. In three other cases, normal, FEA and DCIS components were negative for miR-21 and in the last case all three components were positive. Overall we observed a gradual increase in percentage of miR-21 positive cases from normal, to FEA, DCIS and IDC-NOS. Immunohistochemical staining for PTEN revealed no obvious changes in staining intensities in normal, FEA, DCIS and IDC-NOS. Cytoplasmic staining of PDCD4 increased from normal to IDC-NOS, whereas, the nuclear staining decreased. TM1 staining decreased from positive in normal breast to negative in most DCIS and IDC-NOS cases. In FEA, the staining pattern for TM1 was similar to normal breast tissue.[Conclusions](1) The position relation between FEA and DCIS is closer with the higher grade of FEA. We should take FEA, especially FEA which is high grade and adjacent to DCIS, as a precancerous lesion.(2) Upregulation of miR-21 from normal ductal epithelial cells of the breast to FEA, DCIS and IDC-NOS parallels morphologically defined carcinogenesis. No clear relation was observed between the staining pattern of miR-21 and its previously reported target genes. [Background]BRCA1 associated breast cancer has its own distinctive clinical features, histopathologic character, copy number alterations and gene expression profiles in comparison with sporadic breast cancer and BRCA2 associated cancer. MicroRNAs (miRNAs) play an important role in a series of biopathological functions, such as growth, development, apoptosis, proliferation and carcinogenesis. We aimed to explore the miRNA expression profiles in BRCA1 associated cases compared with BRCA1 non-associated cases.[Methods]MiRNA microarray method was applied to investigate the miRNA expression profiles of BRCA1 associated and non-associated breast cancer cases. Eight BRCA1 mutation cases, ten BRCA1/2 non-mutation sporadic breast cancer cases and 6 normal breast cases (pooled into two samples) were included and compared with 328 flagged miRNAs in our study.[Results]A subgroup of miRNAs was identified between BRCA1 mutation cases and BRCA1 non-mutation cases. With the help of Targetscan software, certain miRNAs were further selected among them, which might play a potential role in BRCA1 pathway.[Conclusions]Compared with BRCA1/2 non-mutation sporadic breast cancer cases and normal cases, BRCA1 mutation cases show a distinctive miRNA expression profiles by hierarchical cluster analysis.更多还原