逆转录聚合酶链反应检测石蜡包埋组织中细胞周期蛋白D1 mRNA对套细胞淋巴瘤的诊断价值

目的 探讨用逆转录聚合酶链反应（RT-PCR）法和竞争性RT-PCR法检测套细胞淋巴瘤（MCL）石蜡包埋组织中细胞周期蛋白D1（cyclin D1）蛋白和mRNA在常规病理工作中的可行性及其诊断和鉴别诊断价值。方法 收集淋巴结内MCL38例、对照组包括结内小B细胞淋巴瘤58例（B小淋巴细胞性淋巴瘤14例,淋巴浆细胞性淋巴瘤3例,滤泡性淋巴34例,淋巴结边缘区B细胞淋巴瘤7例）和淋巴结反应性增生病例20例,用免疫组织化学EnVision法和RT-PCR法、竞争性RT-PCR法检测cyclin D1蛋白及其mRNA的表达,以看家基因PGK作为内对照检测RNA。结果 （1）38例结内MCL中,cyclin D1蛋白阳性率为71．1％（27／38）,对照组均为阴性。（2）116例标本中,可检出内对照PGK基因mRNA表达103例（88．8％）。38例MCL中PGK阳性36例（94．7％）。（3）38例结内MCL中,34例可检出cyclin D1 mRNA表达,去除PGK和cyclin D1 mRNA均阴性的2例,MCL中cyclin D1 mRNA表达的阳性率为94．4％（34／36）。对照组中B小淋巴细胞性淋巴瘤1例检出cyclin D1 mRNA表达,其余病例均未检出cyclin D1 mRNA表达。PCR结果全部经测序证实。（4）用竞争性RT-PCR,38例结内MCL中27例可检出cyclin D1 mRNA高表达,去除2例PGK也为阴性的病例,MCL中cyclin D1 mRNA高表达率为75．0％（27／36）。对照组小B细胞恶性淋巴瘤及淋巴结反应性增生无1例有cyclin D1 mRNA高表达。结论 RT-PCR方法和竞争性RT-PCR方法可在石蜡包埋组织中检测cyclin D1 mRNA的表达,均可用于MCL的诊断。     

套细胞淋巴瘤的临床病理特征和细胞周期蛋白D1在诊断中的意义

目的 观察套细胞淋巴瘤的临床病理学特征及细胞周期蛋白D1染色在诊断中的意义。方法 对8例淋巴结套细胞淋巴瘤作临床病理观察及随访,LSAB法做免疫表型分析（CD45RO、CD5、CD20、细胞周期蛋白D1、Ki-67、bcl-2）。结果 患者年龄43-78岁（平均年龄57岁）,男女3：1。组织学特点为：（1）淋巴结结构破坏并被单一的淋巴样细胞所取代,淋巴细胞以套区增生性、结节性、弥漫性三种模式增生。（2）淋巴样细胞核有一定的不规则性,染色质中等致密,核分裂象少见,类似中心细胞。其中有3例转变为高度侵袭性的母细胞样变型。所有的病例都呈cyclinD1与bcl-2阳性、CD20阳性、CD45RO阴性、CD5阳性。结论 套细胞淋巴瘤有其特征的形态改变及免疫表型。根据组织病理学特征及cyclin D1阳性,可与其它类型的小B细胞淋巴瘤相鉴别。套细胞淋巴瘤的母细胞样变型也应当与其它变型区别。     

细胞周期素D1与套细胞淋巴瘤

肿瘤发生过程中原癌基因的激活是重要因素之一。原癌基因活化的机制有基因扩增、点突变、染色体易位或缺失等〔1〕。在Ｂ细胞淋巴瘤中 ,染色体重排可导致原癌基因的激活而产生过量的癌蛋白 ,使正常的细胞发生癌变〔2〕。现将原癌基因ｂｃｌ 1及其蛋白产物细胞周期素 (ｃｙｃｌｉｎ)Ｄ1与套细胞淋巴瘤的关系综述如下。1　ｃｙｃｌｉｎＤ1结构和功能细胞周期由三类调节因子进行调控 ,它们分别是周期素依赖性激酶 (ｃｙｃｌｉｎ ｄｅｐｅｎｄｅｎｔｋｉｎａｓｅｓ,ＣＤＫｓ)、周期素 (ｃｙ ｃｌｉｎｓ)和周期素依赖性激酶抑制因子 (ｃｙｃｌｉ…     

细胞周期蛋白DI表达、聚合酶链反应及荧光原位杂交检测t（11；14）（q13；q32）易位在套细胞淋巴瘤诊断中的应用

细胞遗传学和分子遗传学的研究证实，70%-80%套细胞淋巴瘤（MCL）中可以检测到t(11:14)(q13:q32)，易位使位于11q13上的bel-1癌基因（也称为PRADI，parathyroid adenomatosis，甲状旁腺腺瘤病或CCND1）处于14q32上1gH基因增强了的调控下，从而被激活，使其编码的细胞周期蛋白（cyclin）D1过度表达。这种易位在其他小     

套细胞淋巴瘤组织细胞周期蛋白D1免疫组织化学染色方法的优化及其在病理诊断中的意义

目前 ,利用常规染色方法 ,大部分实验室使用的细胞周期蛋白 (cyclin)D1抗体虽然能在许多肿瘤 (如乳腺癌 )中准确地标记出相应的蛋白 ,但套细胞淋巴瘤 (mantlecelllymphoma ,MCL)肿瘤细胞的显色多位于胞质和胞膜 ,而事实上cyclinD1应位于核内。为了解决MCL诊断中的难题 ,我们参考倪灿荣介绍的方法[1,2 ] ,对美国Neomarker公司生产的cyclinD1抗体的染色方法进行了探索 ,发现在不同的情况下有不同的着染方式 ,进而摸索出了一套稳定且简单易行 ,针对MCL肿瘤细胞的cyclinD1免疫组织化学染色方法。使其在MCL的诊断和鉴别诊断中发挥出应有的…     

t(11;12)易位伴习惯性流产一例

患者　男 ,33岁。结婚 4年 ,其妻妊娠 5次 ,均在孕 5 0天自然流产。夫妻表型、智力正常 ,身体健康 ,非近亲婚配 ,其妻自诉孕期无有害物质接触史及感染史。夫妻双方性激素检查均正常 ,抗精子抗体阴性 ,Rh血型均为阳性 ,ABO血型测定 :患者为 O型 ,其妻为 B型。其妻优生五项 (弓形虫 ,风疹病毒 ,巨细胞病毒 ,单纯疱疹病毒 、 型 )检查均正常 ,卵泡发育检测良好。患者精液分析正常 ,前列腺液涂片分析正常 ,弓形虫抗体检测阴性 ,解脲脲原体培养阴性。细胞遗传学检查 :常规外周血淋巴细胞培养 ,制备 ,G显带分析 ,计数 5 0个分裂相 ,镜下分析 1…     

一例44,XY,t(13;14),t(13;15)罗伯逊易位

患者　男 ,2 8岁 ,因结婚 4年妻子未孕就诊。首次作精液检查时 ,高倍镜下“可见少数精子 ,均为畸形”。以后多次复查精液结果相同。患者表型正常 ,智力正常。有 2个兄长、5个姐姐 ,均已正常生育 ,本人为家中最小的孩子 ,出生时母亲已 44岁 ,现仍健在。查体 :各系统无异常 ,外生殖器也无异常 ,激素水平也在正常男性范围内。外周血淋巴细胞染色体培养 ,常规 G、C显带 ,油镜下作计数分析与核型分析 ,计数 2 80个分裂相均为 44条染色体 ,核型分析 5 0个 ,均为 44 ,XY,t(13;14) ,t(13;15 )罗伯逊易位核型(图 1) ,易位后的染色体大小、形态与 3…     

套细胞淋巴瘤的临床病理特征及预后因素分析

目的探讨套细胞淋巴瘤（MCL）的临床病理特征及预后因素。方法对102例经形态学及免疫表型检测确定的MCL进行分析,组织病理制片和链霉素抗生物素蛋白过氧化物酶法或EnVision法染色,并进行了随访。结果102例患者中位年龄59岁（30～79岁）,男女之比约2．92：1。淋巴结是最常受累的部位（98／98,100％）,结外常受累的部位：骨髓（29／45,64．4％）、脾脏（36／57,63．2％）、咽淋巴环（15／48,31．3％）、外周血（15／51,29．4％）、肝脏（12／53,22．6％）及胃肠道（15／102,14．7％）;87．7％（71／81）初次就诊时处于临床Ⅲ～Ⅳ期,45．5％（25／55）患者有B症状;48．7％（19／39）患者血清乳酸脱氢酶升高。除7例（6．9％）因组织取材小无法区分病变模式外,余95例中12例（11．76％）为套区增生型,41例（40．2％）结节型,42例（41．2％）弥漫型。75．5％（77）经典型,24．5％（25例）瘤细胞呈母细胞样变型。102例均表达B细胞标记而不表达T细胞标记,96例（94．1％）肿瘤细胞表达细胞周期蛋白D1,70例（71．4％）CD5弱阳性。68例获得随访,中位生存时间10个月（0～89个月）。套区增生型＋结节型、经典型的核分裂象≤15／10HPF,增殖指数≤15％;骨髓无受累,提示患者预后好,而其他临床病理因素对患者生存未见影响。结论国内MCL患者病征与国外患者基本一致,其病变模式、细胞变型、核分裂象、增殖指数、骨髓是否受累及受累程度与预后有关。     

46,XX,t(3p;11p;14q)复杂易位一例

患者　女 ,2 7岁 ,结婚 5年。怀孕 9次 ,均于孕 4 0～ 5 0天自然流产。患者表型正常 ,智力良好。多次妇产科全面检查 ,未发现早期流产的原因。患者母亲无流产史 ,一兄一妹结婚后均正常生育。夫妇外周血淋巴细胞染色体检查 ,丈夫核型正常 ,妻子核型为 4 6 ,XX,t(3;11;14 ) der(3     

两代t(1;2)易位携带者二例

先证者　男 ,4 6岁 ,结婚 2 1年 ,其妻共妊娠 5次 ,现有一男孩 2 0岁 ,表型正常 ,智力稍低。其妻 4次妊娠中均于孕 2 月无明显诱因自然流产。夫妻表型及智力正常 ,非近亲婚配 ,无有毒、有害物质接触史 ,先证者同胞中已婚者均正常生育。细胞遗传学检查 :取外周血常规制备染色体     

滤泡型淋巴瘤中t(14;18)染色体易位的分析及其临床意义

目的探讨滤泡型淋巴瘤（FL）的分子遗传学特征及其在病理诊断中的意义。方法收集55例FL石蜡标本,对照组小B细胞淋巴瘤28例和反应性滤泡增生（RFH）10例,应用套式PCR技术检测FL中,免疫球蛋白重链基因（IgH）的克隆性重排;应用标准PCR技术检测55例FL中t（14;18）易位,以10例RFH做对照;采用双色荧光原位杂交（FISH）技术检测20例淋巴结FL中t（14;18）易位,以4例RFH作为对照;并与PCR检测结果进行比较。结果（1）55例FL中,结内49例,结外6例。男性33例,女性22例,男女比为1．5：1。发病年龄36—79岁（中位年龄57岁）;FL分级：FL1—3分别为25例、19例和11例。（2）55例中50例（90％）检出β-肌动蛋白（actin）,该50例中FR3A阳性24例（48％）,FR2阳性25例（50％）,其中15例（30％）呈FR3A和FR2双阳性,共34例（68％）IgH基因重排。对照组小B细胞淋巴瘤28例中,25例检出β—actin,其中FR3A阳性18例（64％）,FR2阳性17例（61％）,共24例（86％）可检测出克隆性IgH基因重排。4例RFH均未检出IgH基因重排。（3）在44例结内FL中检出15例（34％）t（14;18）易位,其中14例在MBR,1例在mcr。（4）20例中,有16例（80％）可检出t（14;18）易位。结论（1）IgH克隆性重排在FL中的检测率比其他小B淋巴细胞低。（2）FISH检测石蜡包埋组织中t（14;18）易位有助于FL的诊断。FISH比PCR的敏感性更好,操作简便,可用于检测石蜡包埋组织中的分子遗传学改变。     

Cyclin D1 and t(11;14)-positive B-cell neoplasms resembling marginal zone B-cell lymphoma: a morphological variant of mantle cell lymphoma

We describe 3 unusual B-cell non-Hodgkin's lymphomas in which the entire tumors histologically mimicked marginal zone B-cell lymphoma. All patients were male (mean age, 65 years). Excisional biopsy from lymph node (2 of 3) and parotid gland (1 of 3) showed proliferation of monocytoid B-cells with plasmacytoid features (2 of 3) and conspicuous absence of large lymphoma cells (3 of 3). By immunohistochemistry, cyclin D1 was positive (3 of 3), CD23 was negative (3 of 3), and aberrant expression of CD5/CD43 was present in 1 case. Ki67 labeling was greater than 50% in 1 case and 10% to 25% in the other 2 cases. Evidence of the t(11;14) was detectable in all by molecular techniques. One patient died within 15 months, and the other 2 patients had widely disseminated diseases at the last follow-up (8 months). Based on these features, we believed that the best classification for these lesions is the marginal zone B-cell lymphoma-like mantle cell lymphoma.     

弥漫性大B细胞淋巴瘤t(14;18)易位及bcl-2基因扩增的检测

目的探讨弥漫性大B细胞淋巴瘤（DLBCL）t（14;18）染色体异位及bcl-2基因扩增在其分类及临床分期、疗效评估中的作用。方法先对60例DLBCL的标本进行显微切割,获取相对比较纯的肿瘤组织,再使用细胞核芯片荧光原位杂交（FISH）对标本进行t（14;18）易位及bcl-2基因扩增检测,采用免疫组织化学SP法在组织微阵列上同步观测CD20、CD10、bcl-6、MUM1的表达,进行生发中心样（GCB）和非生发中心样（non-GCB）分类;通过病例分析得出治疗效果及临床分期的信息,并统计分析以上各因素之间的关系。结果在60例DLBCL中,10例bcl-2／IgH阳性,18例bcl-2基因扩增;GCB29例（48．3％）,non-GCB31例（51．7％）。经FISH检测t（14;18）阳性10例中,GCB8例,non-GCB2例,差异有统计学意义（P=0．031）。t（14;18）阳性及bcl-2基因扩增的病例bcl-2表达均增高。在36例正规CHOP治疗的病例中,bcl-2扩增13例,无扩增23例,bcl-2扩增的13例之治疗结果显效、部分有效、无效率分别为3例（23．1％）、4例（30．8％）和6例（46．2％）;临床分期情况为Ⅰ-Ⅱ期1例（7．7％）,Ⅲ-Ⅳ期12例（92．3％）,这两项指标与bcl-2不扩增的相比差异均有统计学意义（P=0．019、0．046）。结论t（14;18）易位及bcl-2基因扩增均是引起DLBCL之bcl-2蛋白表达的原因,bd-2阳性者与预后有关的原因难以确定,可能是由于引起其阳性表达的原因不同所致;bcl-2基因扩增与治疗效果较差及临床分期较晚有关;FISH检测t（14;18）染色体易位可用于DLBCL的分类。     

CD5-undetected by immunohistochemistry, t(11;14)(q13;q32)-positive conjunctival mantle cell lymphoma: a case report

Most primary ocular adnexal lymphomas are extranodal marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT). A few cases of ocular adnexal mantle cell lymphomas have been reported in the literature. We present a case of mantle cell lymphoma presenting as conjunctival mass. A 58-year-old man presented with a palpable mass in the left lower tarsal conjunctiva incidentally detected one month previously. Histopathologic examination showed proliferation of monomorphous small-to-medium sized lymphoid cells. On immunohistochemistry, tumor cells were positive for CD20, bcl-2, and cyclin D1, and negative for CD5. PCR analysis for immunoglobulin heavy chain gene rearrangement showed monoclonal B-cell proliferation. t(11;14)(q13;q32), involving the CCND1 and IGH genes, was detected in interphase fluorescent in situ hybridization using formalin-fixed, paraffin-embedded tissue; however, MALT1 gene translocation was not observed. The final diagnosis was mantle cell lymphoma. There was no lymphadenopathy; however, bone marrow involvement of the lymphoma was suspected. The patient has been receiving systemic chemotherapy. This case emphasizes the differential diagnosis of conjunctival mantle cell lymphoma from extranodal marginal zone B-cell lymphomas of MALT regarding the clinical and pathological aspects.     

滑膜肉瘤石蜡包埋组织SYT-SSX融合基因检测的临床病理意义

目的：探讨在石蜡包埋组织中检测SYT－SSX融合基因的可行性及其对滑膜肉瘤的诊断,分型和鉴别诊断的价值。方法：收集滑膜肉瘤标本38例,以恶性周围神经鞘膜瘤,纤维肉瘤,平滑肌肉瘤,尤文肉瘤,血管外皮肉瘤和转移性腺癌作为对照,共40例,均为甲醛固定,石蜡包埋组织,用逆转录－聚合酶链反应（RT－PCR）方法检测SYT－SSX融合基因mRNA表达,以看家基因PBGD作为内对照检测mRNA质量。结果：78例标本中64例（占82．1％）可检出PBGD mRNA表达,38例滑膜肉瘤中33例中可检出SYT－SSX融合基因mRNA表达,对照组无一例检出SYT－SSX基因,去除PBGD及SYT－SSX均阴性病例1例,滑膜肉瘤SYT－SSX融合基因检出率为89．2％（33／37）,33例SYT－SSX阳性滑膜肉中,SYT－SSX1型22例,SYT－SSX2型6例,5例无法区分。融合基因类型与滑膜肉瘤组织学类型有关。10例双相型滑膜肉瘤均为SYT－SSX1型,而18例单相型滑膜肉瘤中SYT－SSX1型12例,SYT－SSX2型6例,二者差异有统计学意义（P＜0．05）,结论：（1）从石蜡包埋组织中检测SYT－SSX融合基因对滑膜肉瘤有较高的敏感性和特异性,可用于滑膜肉瘤的诊断和鉴别诊断;（2）SYT－SSX融合基因类型与滑膜肉瘤组织学类型相关,SYT－SSX2型仅见于单相型。     

SOX11 is useful in differentiating cyclin D1‐positive diffuse large B‐cell lymphoma from mantle cell lymphoma

Hsiao S‐C, Cortada I R, Colomo L, Ye H, Liu H, Kuo S‐Y, Lin S‐H, Chang S‐T, Kuo T U, Campo E & Chuang S‐S
(2012) Histopathology  61, 685–693 SOX11 is useful in differentiating cyclin D1‐positive diffuse large B‐cell lymphoma from mantle cell lymphoma Aims: To characterize the frequency and clinicopathological features of cyclin D1‐positive diffuse large B‐cell lymphoma (DLBCL) and the usefulness of SOX11 in the differential diagnosis from mantle cell lymphoma (MCL). Methods and results: We retrospectively stained 206 consecutive DLBCLs for cyclin D1, and identified three (1.5%) positive cases, comprising two in the elderly with necrosis, and a third with a starry‐sky pattern. All three cases shared the same non‐germinal centre B‐cell (non‐GCB) phenotype [CD5?/CD10?/bcl‐6+/MUM1+/SOX11?], Epstein–Barr virus (EBV) negativity, and absence of CCND1 aberrations by fluorescence in‐situ hybridization. The third case showed both BCL6 and MYC rearrangements: a double‐hit lymphoma. In the same period there were 22 MCLs, all expressing cyclin D1, with 89% cases expressing SOX11, a frequency that is statistically different from cyclin D1‐positive DLBCL. Notably, we identified a pleomorphic MCL initially misdiagnosed as DLBCL. A separate cohort of 98 DLBCL cases was negative for SOX11, with only one case expressing cyclin D1 with a GCB phenotype (CD10+/bcl‐6+/MUM1?). The two patients with tumour necrosis rapidly died of disease. The other two were in complete remission after immunochemotherapy. Conclusions: Cyclin D1‐positive DLBCLs are rare, and they are negative for SOX11 or CCND1 aberration. SOX11 is useful in differentiating cyclin D1‐positive DLBCL from MCL.     

The influence of arsenic trioxide on the cell cycle,apoptosis and expression of cyclin D1 in the Jurkat cell line

Cyclin D1 drives cell cycle progression at the G1/S transition and is believed to play a significant role in tumorigenesis, contributing to efficient proliferation of many cancer cells. Consequently, it is also recognized as an end-point biomarker of therapeutic outcome for different treatment modalities in cancer. In this study we aimed to evaluate the expression and localization of cyclin D1 in arsenic trioxide (ATO) treated Jurkat cells (lymphoblastic leukemia cell line) and to correlate these results with the extent of cell death and/or cell cycle alterations. Jurkat cells were incubated with increasing concentrations of ATO (0.2, 0.6 and 1.0 μM) for 24 h in standard cell culture conditions. To reach our goal we performed annexin V/PI labeling for detection of cell death and RNase/PI labeling for evaluation of cell cycle distribution, which were followed by the respective flow cytometric analyses of ATO-treated Jurkat cells. Transmission electron microscopy was applied for visualization of the cell ultrastructure. For cyclin D1 estimation a biparametric cyclinD1/cell cycle assay was done and localization of the protein was shown after immuno-labeling using light microscopy (ABC procedure) and confocal fluorescence microscopy. We found that there were no significant changes in the percentages of cyclin D1-positive cells after the treatment with ATO, but at the same time mean fluorescence intensity reflecting cyclin D1 content was gradually increasing along with the cell cycle progression, irrespective of the applied dose of the drug. On the other hand, we found a nuclear-cytoplasmic shift of this protein as a major treatment-related response, which was in good accord with an increased rate of cell death and suggested that cyclin D1 cytoplasmic degradation is an important determinant of the therapeutic efficiency of ATO in the Jurkat cell line.