妊娠滋养细胞肿瘤组织中cyclin D1、Rb蛋白产物的表达

目的：探讨增殖相关基因cyclin D1、Rb在妊娠滋养细胞肿瘤中的表达变化。方法：采用免疫组化S－P法,检测20例葡萄胎、15例侵蚀性葡萄胎、15例绒毛膜癌组织中两种基因蛋白产物的表达。结果：在恶性滋养细胞肿瘤组织中,cyclinD1的阳性表达率随临床期别的增高呈递增趋势,而Rb呈递减趋势,二者在Ⅲ期的阳性表达率与Ⅰ期及Ⅱ期相比差异均具有显著性（P＜0．05）;化疗可降低cyclinD1在恶性滋养细胞肿瘤中的阳性表达,而对Rb无影响。结论：cyclin D1的过表达,Rb的缺失在妊娠滋养细胞肿瘤的发展中可能有重要的生物学意义。     

乳腺癌cyclin D1、Rb蛋白表达的意义

目的：探讨cyclin D1蛋白和Rb蛋白的表达状况与乳腺癌发生发展的关系。方法：采用S-P免疫组化法检测52例乳腺癌和20例良性乳腺病变组织中cyclin D1蛋白和Rb蛋白的表达。结果：乳腺癌中cyclin D1过表达率为34．60％（18／52），显著高于良性乳腺病变组织的10．00％（2／20），P〈0．05，cyclin D1过表达出现于导管内癌中并持续于浸润、转移等进展过程中，与患者年龄、肿瘤大小、腋窝淋巴结转移及组织学分级均无相关性。乳腺癌中Rb蛋白阴性表达率为76．91％（40／52），显著高于良性乳腺病变组织的15．00％，P〈0．05；Rb蛋白阴性表达与乳腺癌临床病理特征间无相关性，Rb蛋白表达阴性者中，cyclin D1过表达率为20．00％，而Rb蛋白表达阳性者中，其cyclin D1过表达率为83．33％，两者比较有显著差异（P〈0．05）。结论：cyclin D1蛋白和Rb蛋白与乳腺癌发生、发 展密切相关；cyclin D1、P16及Rb通路失活可能与乳腺癌的发生密切相关。     

Expression and prognostic significance of cyclin B1 and cyclin A in non-small cell lung cancer

Aims:  Aberrant expression of cell cycle regulators has been implicated in the pathogenesis of many neoplasms, including non-small cell lung cancer (NSCLC). The aim was to examine the expression and prognostic value of cyclin B1 and cyclin A, key regulators of the G₂/M checkpoint of the cell cycle, in NSCLC and bronchial precursor lesions.
Methods and results:  Immunohistochemical expression of cyclin B1 and A was examined in 90 cases of stage I–II primary NSCLC and bronchial precursor lesions using tissue microarrays. Increased cyclin B1 and A expression was found in 40.9 and 58.9% of NSCLC cases, respectively, and was significantly higher in primary NSCLC, lymph node metastases and some bronchial precursor lesions compared with normal bronchial epithelium. Increased expression of cyclin A and cyclin B1 correlated with tumour type, poorly differentiated tumours and male gender. A significant association was found between increased cyclin B1 expression and reduced survival using Kaplan–Meier survival analysis. On multivariate analysis, cyclin B1 was not an independent prognostic factor ( P  = 0.067). Cyclin A expression was not associated with survival.
Conclusions:  Cyclin B1 and cyclin A are aberrantly expressed in NSCLC and some precursor lesions. Cyclin B1, but not cyclin A, shows some promise as a potential prognostic marker in NSCLC.     

Analysis of beta-catenin mutations and alpha-, beta-, and gamma-catenin expression in normal and neoplastic human pituitary tissues

The cadherin-catenin system mediates Ca²⁺-dependent cell-cell adhesion, and genetic alterations in these molecules play a significant role in multistage carcinogenesis. Mutations in the β-catenin gene, mostly affecting exon 3, have been detected in malignant cell lines and in primary tumors. Immunohistochemical abnormalities in α-, β-, and γ-catenin have been reported in malignant and benign tumors, and nuclear localization of β-catenin has been associated with mutations in exon 3 of this gene. Mutational analysis of exon 3 of the β-catenin gene was undertaken by polymerase chain reaction (PCR) and sequencing using genomic DNA extracted from frozen tissues, including 4 normal pituitaries, 22 pituitary adenomas, and one pituitary carcinoma. Frozen sections from these cases were used for immunohistochemical detection of β-catenin. We also analyzed immunohistochemical expression of α-, β-, and γ-catenin by paraffin sections from 154 pituitary tumors, including 148 adenomas and 6 carcinomas. Genomic DNA was extracted from paraffin sections of 2 gonadotroph tumors showing nuclear staining for β-catenin and was used for PCR and sequencing of exon 3 of the β-catenin gene. No mutations in exon 3 of the β-catenin gene were found in any of the 23 cases analyzed by PCR and sequencing. In addition, the 2 cases studied by paraffin section immunohistochemistry, with nuclear staining for β-catenin, were negative for mutations in this exon. Normal pituitary expressed all three catenin proteins. Immunostaining usually showed a membranous pattern of reactivity and was generally stronger in normal pituitary than in the adjacent adenomas. Stains for α-catenin were positive in fewer tumors than for β-catenin. The lowest frequency immunopositive tumors and the weakest immunostaining was for γ-catenin. All medically treated prolactinomas were negative for γ-catenin, whereas treated growth hormone adenomas were less often positive for both α- and γ-catenin than for untreated tumors. The percentage of positive cases for β-catenin was the same in these two groups. Most pituitary carcinomas were negative for both α- and γ-catenin but were β-catenin positive. These results indicate that (i) mutations in exon 3 of the β-catenin gene are uncommon in pituitary tumors, and (ii) expression of α-, β-, and γ-catenin is decreased in pituitary adenomas compared to normal pituitary tissues.     

Different patterns of beta-catenin expression in gastric carcinomas: relationship with clinicopathological parameters and prognostic outcome

AIMS: The cadherin-catenin complex is known to play a critical role in maintenance of cell adhesion. Additionally beta-catenin (beta-ct) can also take part in signal transduction and nuclear beta-ct expression could be correlated with poor prognosis in several malignancies. Since, in gastric cancer, this role of beta-ct is still uncertain, we investigated the expression pattern of beta-ct as well as the possible prognostic role. METHODS AND RESULTS: beta-catenin expression was immunohistochemically investigated in a retrospective series of 401 R0-resected gastric carcinomas. Out of these cases, 54 tumours (13.5%) revealed a preserved membranous beta-ct expression similar to that in normal gastric mucosa. In 80 tumours beta-ct expression was moderately reduced and in 117 tumours highly reduced. In 150 tumours (37.4%), no or only a weak membranous beta-ct expression was found. Additionally, in 53 tumours, a strong beta-ct expression could be observed in the cytoplasm with a simultaneous nuclear beta-ct immunoreactivity in 17 of these 53 tumours, while nine tumours only showed nuclear immunoreactivity without cytoplasmic staining. There were no significant correlations between the degree of membranous beta-ct expression or the different staining pattern (membranous vs. cytoplasmic/nuclear) and the grade of tumour differentiation, the histological tumour type according to Lauren, as well as with the prognostic parameters pT, pN category and vascular invasion. No associations could be found with tumour cell proliferation and the expression of E-cadherin, irrespectively of the different beta-ct staining pattern. Univariate analysis revealed no influence on survival, either for membranous or for cytoplasmic/nuclear beta-ct expression. CONCLUSION: Our data on 401 tumours suggest that activation of the Wnt/beta-catenin signalling does also occur in a subset of gastric carcinomas. However, in gastric cancer, neither the presence of cytoplasmic/nuclear beta-ct expression nor the reduction or loss of membranous beta-ct expression is correlated with a specific histological tumour type, tumour progression or prognosis.     

结直肠癌中cyclin D1和p27蛋白的表达及其意义

目的　研究cyclinD1和p2 7蛋白在结直肠癌发生、发展中的作用及其与结直肠癌临床病理特征关系。 方法　收集5 8例手术切除的结直肠癌标本 ,同时取距癌灶 >5cm的正常组织 ,应用免疫组化S P法检测癌组织及正常组织中cyclinD1和p2 7蛋白的表达。结果　cyclinD1蛋白在结直肠癌的表达阳性率为 5 5 17%,正常组织无表达 (P <0 0 1) ;cyclinD1蛋白的表达阳性率在 6 0岁以上年龄组高于 6 0岁以下年龄组 (P <0 0 5 ) ;cyclinD1蛋白的表达与肿瘤组织分化程度负相关 (P <0 0 1)。p2 7蛋白在结直肠癌的表达阳性率为 5 5 17%,在结直肠正常组织的表达阳性率为 96 5 5 %(P <0 0 1) ;p2 7蛋白的表达与肿瘤组织分化程度负相关 (P <0 0 1)。cyclinD1和p2 7蛋白在结直肠癌的表达呈正相关 (r =0 5 82P <0 0 1)。 结论　cyclinD1蛋白过表达与 p2 7蛋白失活可加速细胞周期转化 ,促进结直肠癌的发生 ,cyclinD1和 p2 7蛋白的检测可作为评价结直肠癌恶性程度和判断预后的重要指标。     

Cyclin D1 in invasive breast carcinoma: favourable prognostic significance in unselected patients and within subgroups with an aggressive phenotype

Aims: To study the clinicopathological and prognostic value of cyclin D1 overexpression in patients with breast carcinoma. Methods and results: Immunohistochemistry was performed on paraffin‐embedded tissue specimens from 290 invasive breast carcinomas to detect the proteins cyclin D1, oestrogen receptor (ER), progesterone receptor (PR), p53, c‐erbB2, and topoisomerase IIα (topoIIα). Cyclin D1 staining was quantified using a computerized image analysis method. Cyclin D1 overexpression characterized smaller, ER‐positive and PR‐positive tumours (P = 0.017, P < 0.0001, and P < 0.0001, respectively), of a lower histological and nuclear grade (P = 0.011 and P < 0.0001, respectively), and with reduced expression of topoIIα (P = 0.001) and p53 (P < 0.001). Cyclin D1 was found to have an independent favourable impact on the overall survival of both the unselected cohort of patients (P = 0.011) and of patients with ER‐negative and lymph node‐positive tumours (P = 0.034 and P = 0.015, respectively). In triple‐negative tumours, cyclin D1 overexpression was found to have independent favourable impacts on both overall and relapse‐free survival (P = 0.002 for both). Conclusions: This is the first immunohistochemical study to dissociate the advantageous prognostic effect of cyclin D1 overexpression from its association with ER expression, and to provide evidence that cyclin D1 overexpression may be a marker of prolonged survival in patient subgroups with aggressive phenotypes.     

大鼠海马tau蛋白过度磷酸化时细胞周期蛋白cyclin D1的表达

目的:探讨大鼠海马tau蛋白过度磷酸化时细胞周期蛋白cyclin D1的表达.方法:用cAMP-依赖性蛋白激酶 A (PKA) 的激动剂Forskolin注射大鼠侧脑室,免疫印迹和免疫组织化学技术检测大鼠海马tau蛋白的磷酸化水平,同时采用免疫印迹、免疫组织化学和RT-PCR方法检测海马cyclin D1和cyclin D1 mRNA.结果:侧脑室注射Forskolin 48h,大鼠海马tau蛋白Ser214、Ser396和Ser202/Thr205位点的磷酸化水平升高,同时检测到细胞周期蛋白cyclin D1和cyclin D1 mRNA.结论:侧脑室注射Forskolin诱导大鼠海马 tau蛋白过度磷酸化的同时,大鼠海马出现细胞周期蛋白cyclin D1的表达,提示了tau蛋白过度磷酸化与细胞周期相关蛋白cyclin D1之间可能的内在联系.     

Expression and amplification of Her2, EGFR and cyclin D1 in breast cancer: immunohistochemistry and chromogenic in situ hybridization

Determination of Her2, epidermal growth factor receptor (EGFR) and cyclin D1 status is now of major clinical importance due to the development of molecule-targeting drugs in anticancer therapy. Immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH) are the most simple and convenient methods for evaluating gene alterations and their protein consequences. The purpose of the present study was to investigate the status of Her2, EGFR and cyclin D1 on both IHC and CISH in 95 primary breast carcinomas. There was substantial consistency between the IHC and CISH results of Her2 and EGFR, showing fair agreement between protein overexpression and gene amplification. However, cyclin D amplification was not related to protein overexpression. Moreover, there was no correlation between Her2, EGFR and cyclin D1. Her2 protein overexpression and amplification were positively associated with histological grade, nuclear grade and inversely correlated with the expression of estrogen receptor (ER) and progesterone receptor (PR). In ER-negative and postmenopausal patients, EGFR gene amplification was strongly associated with worse recurrence-free survival (P = 0.0087, P = 0.0149, respectively). Overall, the present findings suggest that EGFR gene amplification is important in predicting prognosis and this should be evaluated in breast carcinoma in addition to Her2 status in routine pathological practice.     

食管癌及癌旁组织中EGR-1、c-fos、cyclin D1的表达

目的：研究ＥＧＲ－１、ｃ－ｆｏｓ、ｃｙｃｌｉｎＤ１３种基因ｍＲＮＡ及其相应蛋白在人食管癌癌变过程中的表达水平及其相关性,并探讨其与食管癌的发生、发展及生物学行为的关系。方法：原位杂交法检测４７例食管鳞状细胞癌及其癌旁、上切缘中３种基因的ｍＲＮＡ、免疫组化ＳＡＢＣ法检测蛋白表达水平。结果：３种基因原位杂交、免疫 化阳性表达产物均分别定位于胞浆及胞核中,ＥＧＲ－１ｍＲＮＡ、ｃｆｏｓｍＲＮＡ、ｃｙｃｌｉ     

Mantle cell lymphoma in Taiwan: clinicopathological and molecular study of 21 cases including one cyclin D1-negative tumor expressing cyclin D2

Mantle cell lymphoma (MCL) is a rare B-cell lymphoma that has never been characterized in Taiwan. The purpose of the present paper was to retrospectively identify 21 cases in male patients, with a median age of 61, involving lymph node (91%), marrow (71%), and peripheral blood (23%). Eighteen (86%) were in stages III/IV with 1 and 5 year survival rates of 78% and 17%, respectively. Mixed nodular and diffuse pattern (45%) was most common while interstitial pattern (92%) predominated in marrow. Eighteen (86%) were of classical morphology, two were pleomorphic and one was blastic. The tumors expressed IgM and bcl-2 (100%), cyclin D1 (95%), CD5 (86%), CD43 and IgD (62%), CD52 (60%), and bcl-6 (5%). Ki-67 index>or=30% (P=0.1834) was associated with a trend toward poorer survival while p21, p27, or p53 expression was not statistically significant for survival. Real-time polymerase chain reaction for cyclin D1 (CCND1) gene mRNA expression showed high levels in nine cyclin D1-positive patients and a low level in the single cyclin D1-negative patient. The latter patient was cyclin D2 positive and negative for immunoglubuin heavy chain gene and CCND1 gene translocation by locus-specific interphase fluorescent in situ hybridization. In conclusion, it is confirmed that the usual morphological variants and aberrant immunophenotype of MCL in the West occur in Taiwan and that this disease carries a poor prognosis.     

Alterations of expression of Rb,p16INK4A and cyclin D1 in non-small cell lung carcinoma and their clinical significance

Inactivation of the Rb pathway in non-small cell lung carcinoma (NSCLC) occurs mostly through inactivation of the cyclin-dependent kinase inhibitor p16^INK4A and/or up-regulation of cyclin D1. In order to assess the frequency and the prognostic value of these abnormalities in NSCLC, immunohistochemical analysis of Rb, p16^INK4, and cyclin D1 has been performed on 168 cases of NSCLC including 77 squamous cell carcinomas, 43 adenocarcinomas, and 48 basaloid carcinomas. The reduced survival rate of basaloid carcinoma (stage I–II) compared with other histological types of NSCLC was confirmed (p = 0·008). Loss of protein expression of Rb and p16^INK4A was observed in 12 per cent and 58 per cent of NSCLC cases respectively and cyclin D1 overexpression in 43 per cent. There was an inverse correlation between Rb and p16 expression ( p < 0·0001) and a direct correlation between Rb and cyclin D1 expression ( p = 0·0007). In univariate analysis, Rb-negative adenocarcinomas at stages I–II had a significantly shorter survival than Rb-positive cases ( p = 0·04) and stages I–II p16-positive cases had a shorter survival than p16-negative cases ( p = 0·02), which was more significant in basaloid carcinoma ( p = 0·003). p16 status retained its influence on survival in multivariate analysis at stage I–II for all cases ( p = 0·01) and for basaloid carcinoma ( p = 0·005). Cyclin D1 overexpression did not influence survival. Combined Rb/p16/cyclin D1 phenotypes in univariate analysis showed a shorter survival for Rb-negative/p16-positive/cyclin D1-negative tumours ( p = 0·002). These results, linked to previous data, indicate that the Rb pathway of G1 arrest is initially disrupted in the vast majority of NSCLCs (83 per cent), but could not confirm an unfavourable role for each individual event (p16^INK4A loss or cyclin D1 up-regulation) in prognosis. Copyright © 1999 John Wiley & Sons, Ltd.     

High-level cytoplasmic cyclin D1 expression in lymph node metastases from prostate cancer independently predicts early biochemical failure and death in surgically treated patients

Fleischmann A, Rocha C, Saxer‐Sekulic N, Zlobec I, Sauter G & Thalmann G N
(2011) Histopathology 58 , 781–789
High‐level cytoplasmic cyclin D1 expression in lymph node metastases from prostate cancer independently predicts early biochemical failure and death in surgically treated patients Aims: To test the prognostic significance of cyclin D1 in nodal‐positive prostate cancer. Methods and results: Nuclear and cytoplasmic cyclin D1 expression was evaluated in 119 nodal‐positive prostate cancer patients undergoing radical prostatectomy and extended lymphadenectomy. Cyclin D1 was correlated with various tumour features and biochemical recurrence‐free survival (bRFS), disease‐specific survival (DSS) and overall survival (OS). In the metastases, high‐level cytoplasmic cyclin D1 expression independently predicted poor outcome (5‐year bRFS, 12.5% versus 26.4%, P = 0.006; 5‐year DSS, 56.3% versus 80.7%, P = 0.007; 5‐year OS, 56.3% versus 78.7%, P = 0.011). These patients had a 2.62‐fold elevated risk of dying from prostate cancer as compared with patients with low‐level cytoplasmic cyclin D1 expression (P = 0.024). All other subcellular compartments of cyclin D1 expression in primary tumours and metastases were prognostically non‐significant. Conclusions: The subcellular location of cyclin D1 expression in prostate cancer is linked to specific clinical courses. Survival stratification according to biomarker expression in metastases indicates an important role for tumour sampling from these tissues.     

Evaluation of cyclin D1 expression and its subcellular distribution in mouse tissues

Cyclin D1 is a key cell-cycle regulatory protein required for the cell to progress through G1 to S phase. We have shown by Western blot analysis that cyclin D1 has a wide distribution in adult mouse tissues, with its level of expression being tissue-dependent. Immunohistochemistry has also shown that cyclin D1 may be present in the cytoplasm, in the nucleus or in both these cell compartments: cytoplasmic staining was observed in both proliferating cells (e.g. kidney, intestine, stomach and salivary gland) and in the non-dividing cells (the mature neurons of adult brain), while nuclear staining was seen in the neurons of the embryonic nervous system. Immunoelectron microscopy results indicate that, in tissues where cyclin D1 is present in both compartments (e.g. intestinal enterocytes), it may move via nuclear pores from the nucleus to the cytoplasm, and vice versa. The findings as a whole suggest that cyclin D1 may play multiple roles within specific tissues, probably by interacting with different substrates, and that its transit between nuclear and cytoplasmic compartments may help maintain cell homeostasis.     

c-Myc、SIRT1、acetyl-p53在乳腺癌中的表达及其对预后的影响

目的：探讨c-Myc、SIRT1及acetyl-p53蛋白在乳腺癌中的表达及其对预后的影响。方法：应用免疫组织化学方法检测90例乳腺癌和30例乳腺增生症中的c-Myc、SIRT1（Sirtuin type 1）、acetyl-p53蛋白表达,结合临床病理资料和随访资料,进行预后分析。结果：c-Myc阳性组乳腺癌患者的5年无瘤生存率和5年总生存率（分别为5 9.0%/6 7.2%）低于c-M y c阴性组（分别为8 6.2%/8 6.2%）,S I RT 1阳性组乳腺癌患者的5年无瘤生存率和5年总生存率（分别为5 6.1%/6 4.9%）低于S I RT 1阴性组（87.9%/87.9%）;acetyl-p53阳性组乳腺癌患者的5年无瘤生存率和5年总生存率（分别为86.7%/86.7%）高于acetyl-p53阴性组（分别为58.3%/66.7%）,差异均有统计学意义（P〈0.05）。结论：c-Myc及SIRT1蛋白高表达的乳腺癌患者预后差,而acetyl-p53蛋白高表达的乳腺癌患者预后好。     

结肠肿瘤中wisp-1的表达及意义

目的探讨wisp-1在原发性结肠癌组织中的表达及临床意义。方法应用免疫组化EnVision两步法检测wisp-1蛋白在138例结肠癌组织(48例含癌旁组织)、37例结肠腺瘤组织及66例结肠正常黏膜组织中的表达;应用实时荧光定量PCR检测wisp-1基因在37例新鲜原发性结肠肿瘤标本及对照组37例正常结肠中的表达;分析wisp-1蛋白、基因表达与患者临床病理特征的关系。结果 wisp-1蛋白在正常结肠、结肠腺瘤、癌旁组织及结肠癌中阳性率分别为53.0%、67.6%、75.0%、76.1%,其在结肠癌及癌旁组织中表达高于正常结肠(P<0.05);wisp-1蛋白表达与淋巴结转移及浸润深度有关(P<0.05);wisp-1基因在结肠肿瘤中平均表达是正常结肠中的1.349倍(P>0.05)。结论 wisp-1蛋白异常可能在结肠癌的发生过程中起重要作用,而其基因在结肠癌中的作用尚需进一步研究。     

Expression of cyclin D1 and c-Ki-ras gene product in human epithelial ovarian tumors

The expression of cyclin Dl gene product in human ovarian tumors was studied. We found that cyclin D1 is expressed at high levels in several ovarian cancer cell lines. Immunohistochemical study also showed that a significant proportion of primary ovarian tumor tissues overexpressed cyclin D1 gene product. Clear nuclear staining of cyclin Dl protein was detected in 28% of the cases. We also characterized the expression of c-Ki-ras gene product in ovarian cancer cell lines and tumor tissues. Amplification or overexpression of this protooncogene has been reported in ovarian tumors from Taiwan. These results show that c-Ki-ras is strongly expressed in PA-1 and NIH: OVCAR-3 cells in which cyclin D1 also expressed at high levels. Specific cytoplasmic staining of c-Ki-ras protein was detected in 11 tumors (52%). Statistical analyses show a strong positive correlation between cyclin D1 and c-Ki-ras immunoexpression. Thus, these data support the ideas that cyclin D1 may be involved in the pathogenesis of ovarian cancer, and coactivation of cyclin D1 and c-Ki-ras gene expression may represent one of the major pathways that lead to the development of ovarian cancer in Taiwan.