Prognostic value of DNA ploidy in 653 Japanese women with node-negative breast cancer

Background. The prognostic value of DNA ploidy has been extensively studied in breast cancer; however, the results remain controversial. Flow cytometry (FCM) studies of DNA ploidy on frozen sections have not yet been performed in a large series of Japanese women with node-negative breast cancer. Methods. An FCM analysis of DNA ploidy was performed on frozen sections of node-negative breast cancer from 653 Japanese women, with a mean follow-up duration of 46 months. Results. Three hundred and twenty-four (49.6%) patients showed a diploid tumor, while 329 (50.4%) showed an aneuploid tumor. There was a significant correlation between DNA ploidy and estrogen receptor (ER) status. Patients with an aneuploid tumor had a significantly worse outcome in terms of both disease-free survival (DFS; P = 0.0116) and overall survival (OS; P = 0.0492) than those with a diploid tumor, while the same effect, in terms of DFS, was also seen in ER-positive breast cancer. Multivariate analyses indicated DNA ploidy to be an independent prognostic factor for DFS, while DNA ploidy and tumor size were found to be independent prognostic factors for OS. DNA ploidy was also shown to be an independent prognostic factor for DFS in ER-positive breast cancer. Conclusion. Our findings demonstrated DNA ploidy, based on FCM, to have an important prognostic value in Japanese women with node-negative breast cancer. Received: March 23, 2001 / Accepted: June 6, 2001     

前列腺癌的细胞核DNA图像分析研究

目的探讨DNA图像分析技术在前列腺癌(PC)诊断中的应用价值。方法应用计算机图像分析技术测定对5例正常前列腺(NP)和30例PC的标本DNA倍体。结果5例NP中均为整倍体(2C和3~4C),30例PC中有23例出现非整倍体,并且随着PC分级的增高,其倍体率也增大。结论细胞核DNA含量和倍体测定是反映前列腺癌细胞增殖能力的重要生物学定量指标,能较客观反映PC的预后,为正确诊断恶性肿瘤提供依据。     

Prognostic significance of DNA ploidy in patients with stage II colorectal cancer

DNA ploidy status associated with colorectal cancer has been investigated widely in recent years. But the relationship between DNA analysis and prognosis has not been confirmed.[1(3] Some investigations showed that DNA aneuploid status was associated with lymph node involvement and poor differentiation, and thus may predict a poor clinical outcome in patients with colorectal cancer. These observations suggest that DNA ploidy abnormality may influence the development and progression of colo…     

细胞DNA倍体图像分析在宫颈癌及癌前病变筛查中的应用

目的利用显微图像分析技术对宫颈脱落细胞做DNA倍体检测、分析,以探讨该技术在宫颈癌及癌前病变筛查中的应用价值。方法对619例宫颈脱落细胞样本分别采用常规细胞学镜检和细胞DNA倍体分析法作检测,其中阳性样本患者行组织活检,并将3种检测结果进行对比分析。结果619例样本,常规细胞学镜检共检出ASC—US42例、ASC—H6例、LSIL29例、HSILl8例;采用细胞倍体分析法检出DI≥2．5时,可见DNA倍体异常细胞（1～2个）的样本102例、DNA倍体异常细胞（≥3个）的样本41例、DNA倍体异常细胞（≥25个）的样本14例;48例组织活检样本检出炎症7例、CINI20例,CINⅡ13例、CINⅢ2例、IC6例。CINⅡ～Ⅲ对应TBS系统中的HSIL计算灵敏度为81．0％;对应SIL和对应ASC＋SIL的灵敏度分别为85．7％和100％。HSIL与CINⅡ～Ⅲ比较有非常显著的统计学意义。CINⅡ～Ⅲ对应DNA倍体异常细胞（≥25个）计算灵敏度为66．7％;对应可见DNA倍体异常细胞（≥3个）的灵敏度为100％。DNA倍体异常细胞（≥25个）与CINⅡ～Ⅲ比较也有非常显著的统计学意义。结论细胞DNA倍体图像分析法在宫颈癌及癌前病变筛查中有较高的灵敏度,若联合常规宫颈细胞学检查,可明显提高宫颈癌筛查的质量控制水平。     

组织原位分析鼻咽癌细胞核DNA干系倍体及其异质性

组织原位分析鼻咽癌细胞核的DNA干系倍体及其异质性。收集42例鼻咽癌病例的归档蜡块,4μm、8μm连续组织学切片各一张。每个病例选取三个癌巢或肿瘤组织区域(共126个样本),使用TIGER细胞图像分析仪4μm组织切片测量鼻咽癌细胞核DNA的光密度等参数,8μm组织切片测量单个完整鼻咽癌细胞核的体积等参数,计算获得每个癌巢或肿瘤组织区域的以单个完整鼻咽癌细胞核的体积为单位的DNA含量(以体积积分光密度表述),以同一切片内正常鼻咽部上皮非基底细胞作为其内参照,计算其DNA干系倍体。结果显示:鼻咽癌细胞核的DNA干系倍体主要集中在1.76～3.00(DNA指数为0.88～1.50)之间,14个样本为DNA干系四倍体,仅2个样本为DNA干系亚二倍体;DNA干系倍体异质性率为66.67%。可得结论:(1)鼻咽癌DNA干系倍体主要在近二倍体与近四倍体之间;(2)鼻咽癌中存在DNA干系倍体异质性。     

Automated assessment of DNA ploidy,chromatin organization,and stroma fraction to predict prognosis and adjuvant therapy response in patients with stage II colorectal carcinoma

DNA ploidy, tumor stroma, and chromatin organization have important implications in tumorigenesis and patient outcome. Automated image cytometry tools were developed to quantitatively measure DNA ploidy (P), stroma fraction (S), and chromatin organization or Nucleotyping (N). This study aimed to discover their clinical value in different stages of colorectal cancer (CRC) in a Chinese patient population. A total of 496 CRC patients of stages I, II, and LMCRC (liver metastatic CRC) were enrolled in this study. Stage II CRC patients with diploidy, low-stroma, or chromatin homogenous status predicted significantly higher 5-year OS and DFS. We constructed a PSN-panel enabled the stage II patients to be further stratified into low-, middle-, high-risk groups, the 5-year OS (89.5% vs 67.9% vs 60.9%, P<0.001) and DFS (86.0% vs 62.3% vs 53.6%, P<0.001) were stratified significantly. In addition, when combined the PSN-panel with T stage or MSS status in stage II patients, the PSN-low risk patients showed significant longer 5-year OS and DFS than the PSN-high risk patients in T3 (OS: 86.3% vs 65.3%, P=0.015; DFS: 83.5 vs 59.8%, P=0.013) or MSS (OS: 86.4% vs 63.9%, P=0.005; DFS: 85.5 vs 57.8%, P=0.003) patients. Finally, in the group of stage II patients with at least one high-risk factor (non-diploidy, high-stroma, chromatin heterogenous), patients who received adjuvant therapy showed significantly longer OS (72.1% vs 48.3%, P=0.007) and DFS (64.5% vs 43.9%, P=0.015) than those who did not receive adjuvant therapy. In contrast, P, S, N couldn’t predict the prognosis of stage I and LMCRC patients. Overall, our data demonstrate that the PSN panel is an accurate prognostic tool that can guide treatment decisions for Chinese stage II CRC patients.     

Combined flow cytometry determination of S-phase fraction and DNA ploidy is an independent prognostic factor in node-negative invasive breast carcinoma: analysis of a series of 271 patients with stage I and II breast cancer

Purpose. To assess the significance of S-phase fraction (SPF) and DNA ploidy evaluated by DNA flow cytometry as prognostic markers in stage I or II breast cancer.Patients and methods. A series of 271 patients, treated by surgery, radiotherapy ± systemic therapy was analyzed (median follow up: 64 months). Standardized flow cytometry cell preparation from frozen samples and consensus rules for data interpretation were followed. Three SPF classes were defined on the basis of tertiles after adjustment for ploidy. Four groups were defined based on combinations of DNA ploidy (DIP: diploid; ANEUP: aneuploid) and SPF: DIP and low SPF (DL, n=37), DIP and medium or high SPF (DMH, n=76), ANEUP and low SPF (AL, n=24), ANEUP and medium or high SPF (AMH, n=68). Local control rate (LCR), disease-free survival (DFS), metastasis-free survival (MFS), and overall survival (OS) were correlated with DNA ploidy, SPF, DL to AMH groups, T and N stages, SBR grading, age, and hormonal status on univariate and multivariate analysis (Cox model).Results. On univariate analysis, DFS and LCR were higher for DIP tumours. High SPF values were associated with shorter DFS. LCR, MFS, DFS, and OS rates were significantly different with an increasingly poorer prognosis from DL to AMH. On multivariate analysis, groups DL to AMH, histological node involvement and T stage were independently associated with MFS, and DFS. In N(–) patients, DL to AMH remained independent for MFS and DFS. For SBR III tumours, MFS and OS were significantly different in DL to AMH groups. These results strongly support the use of combined evaluation of DNA ploidy and SPF as independent parameters in clinical trials for N(–) stage I and II breast cancer.     

Prognostic value of ploidy of primary tumour and nodal secondaries in colorectal cancers

Tumour ploidy is of prognostic value in colorectal cancer, DNA aneuploid tumours having a worse outlook. Nearly all studies have concentrated on the DNA content of the primary tumour. We have examined the ploidy of the primary tumour and its lymph node metastases in 71 cases of Dukes' stage C disease, to see whether this provides greater prognostic information than the primary alone. Analysis was performed using formalin-fixed, paraffin-embedded tumour sections. Ploidy of primary and metastases was different in 20 cases (28%), aneuploid nodes being seen with diploid primaries and vice versa. Ploidy of both the primary (χ² = 4.86, P = 0.03) and secondary (χ² = 4.86, P = 0.03) tumours predicted survival in univariate analysis. Combining the ploidy of primary and nodes, three prognostic groups could be defined — diploid primaries with diploid metastases (hazard relative to both aneuploid, 0.36) had significantly better survival than cases where the ploidy of the primary and nodes were mixed (relative hazard 0.47–0.56), which did better than cases with aneuploid primary and nodes. This study demonstrates that ploidy variation between primary and secondary tumours is common, and better prognostic information may be gained by studying both.     

Prognostic impact of DNA ploidy pattern,S-phase fraction (SPF), and proliferating cell nuclear antigen (PCNA) in patients with primary gastric lymphoma

BACKGROUND: DNA ploidy, S-phase fraction (SPF), and proliferating cell nuclear antigen (PCNA) are considered to be significant prognostic factors in non-Hodgkin lymphomas. However, reports on their prognostic importance in gastric lymphoma patients are relatively lacking. METHODS: In the present study, we retrospectively studied the above-mentioned parameters in 29 patients with primary gastric lymphoma; 11/29 had B-low grade mucosa associated lymphoid tissue lymphoma (B-MALT), while 18/29 had diffuse large B-cell lymphoma (DLBCL), according to WHO classification. Proliferative activity was studied by staining against PCNA; in addition, the prognostic significance of DNA ploidy and SPF, as determined by flow cytometry, were investigated and compared to the results of the PCNA stainings. RESULTS: Seven out of 29 patients were found to have aneuploid tumors; DNA index values were >1 for all aneuploid lymphomas. There was no difference in DNA aneuploidy in MALT vs. DLBCL. The mean percentage of SPF was 11.4. SPF was found significantly lower in MALT vs. DLBCL (P < 0.05). The mean percentage of PCNA positive tumor cells was 52.6. PCNA protein expression was significantly lower in MALT vs. DLBCL (P < 0.0001). There was a significant positive correlation between PCNA score and SPF (P < 0.01, by Spearman analysis). DNA ploidy had no impact on survival in the present study. Both SPF and PCNA expression were important prognostic factors in the univariate analysis; however, in the multivariate analysis, the only independent prognostic factor for survival was PCNA expression. CONCLUSIONS: These findings indicate that SPF and PCNA are significant prognostic factors in patients with primary gastric lymphomas. However, in the present study, DNA ploidy had no impact on survival in patients with primary gastric lymphomas.     

DNA ploidy and S-phase fraction in medullary carcinoma of the breast -a flow cytometric analysis using archival material

Summary In a population of 110 primary breast cancers with medullary features, registered in the Danish Breast Cancer Cooperative Group (DBCG) from 1977-82, we have determined ploidy and S-phase fraction (SF) by flow cytometry (FCM) on paraffin embedded tumour tissue. The distribution of DNA ploidy is not different from the distribution described for breast cancers in general. No difference is found between the subgroups of medullary and non-medullary cancer when using a new simplified histopathological definition of medullary carcinoma of the breast, recently proposed by us. When using the definition proposed by Ridolfiet al. in 1977, we find significantly more tumours with aneuploidy and high SF in the groups of typical medullary carcinoma (TMC) and atypical medullary carcinoma (AMC) than in the small group of non-medullary carcinoma (NMC), which seems a paradox, as patients with NMC have the worst prognosis. However, the number of patients in the NMC group is very small, and the percentage of aneuploid tumours is very low. In 84 protocolled patients we found no statistically prognostic importance of ploidy or SF, either in the whole group assessed or when stratifying for the histopathological subgroups. However, a prognostic influence of SF can be traced for the non-medullary cancers, according to the new definition, but not for the medullary cancers of the breast. The result emphasizes the impression of MC as being biologically different from other histological types of breast cancer.     

DNA ploidy and S-phase fraction of neoplastic and non-neoplastic lesions of the human gallbladder

Studies on the cell kinetics of the human gallbladder are difficult because of epithelial degeneration by bile. Using the epithelial isolation technique, however, we were able to determine the degree of degeneration and to examine the cell kinetics of gallbladder lesions in freshly resected surgical specimens. Normal and neoplastic epithelial were isolated nonenzymatically from freshly resected gallbladder. The nuclear DNA content and S-phase fraction were estimated in 110 patients with gallbladder lesions by flow cytometry (FCM). Normal tissues and all lesions except carcinomas were diploid. The S-phase fraction of gallstone cases was significantly higher (1.47 ± 0.70%; mean ±SD) than normal (0.79 ± 0.39%) (P <0.0006). All gallbladder carcinomas were multiploid, and their S-phase fraction was 11.63 ± 3.65%. Cell renewal of normal gallbladder is low. In the gallstone cases, the S-phase fraction was increased, possibly correlated with carcinogenesis. © 1996 Wiley-Liss, Inc.     

横纹肌肉瘤(RMS)DNA倍体与临床病理关系的研究

本文应用ＥＭＡＩＬ－１００病理图像分析仪（ｉｍａｇｅ－ａｎａｌｙｓｉｓ，ＩＡ）对ＲＭＳ瘤细胞的ＤＮＡ倍体含量进行测试，以揭示ＤＮＡ倍体含量与ＲＭＳ的诊断、组织类型、分级、ＰＣＮＡ表达强度及预后的关系。通过对确诊的４３例ＲＭＳ进行测试，发现所有患者的瘤细胞ＤＮＡ异倍体含量（包括多倍体和非整倍体）显著高于其二倍体含量，而在ＲＭＳ各组织分级、ＰＣＮＡ表达强度（尽管它们与ＤＮＡ异倍体含量呈正相关）及类型之间瘤细胞的ＤＮＡ倍体分布并无显著性差异（ｂＲＭＳ除外）。说明ＤＮＡ倍体含量在ＲＭＳ的诊断上有参考价值，而对其组织类型、分级、ＰＣＮＡ表达强度及预后意义不大。     

Clinical application of DNA ploidy to cervical cancer screening: A review

Screening for cervical cancer with DNA ploidy assessment by automated quantitative image cytometry has spread throughout China over the past decade and now an estimated 1 million tests per year are done there. Compared to conventional liquid based cytology, DNA ploidy has competitive accuracy with much higher throughput per technician. DNA ploidy has the enormous advantage that it is an objective technology that can be taught in typically 2 or 3 wk, unlike qualitative cytology, and so it can enable screening in places that lack sufficient qualified cytotechnologists and cytopathologists for conventional cytology. Most papers on experience with application of the technology to cervical cancer screening over the past decade were published in the Chinese language. This review aims to provide a consistent framework for analysis of screening data and to summarize some of the work published from 2005 to the end of 2013. Of particular interest are a few studies comparing DNA ploidy with testing for high risk human papilloma virus (hrHPV) which suggest that DNA ploidy is at least equivalent, easier and less expensive than hrHPV testing. There may also be patient management benefits to combining hrHPV testing with DNA ploidy. Some knowledge gaps are identified and some suggestions are made for future research directions.     

Flow cytometric DNA analysis of fresh prostatic resections: Correlation with conventional prognostic parameters in patients with prostate cancer

Background. DNA ploidy analysis has been investigated as a prognostic indicator in prostate cancer. Most of the data is derived from retrospective studies using paraffin-embedded tissue. This method has drawbacks related to the quality of DNA histograms and uncontrolled data collection. Methods. DNA ploidy analysis of freshly resected prostatic tissue was prospectively compared with conventional prognostic variables in 97 men treated with radical prostatectomy for localized prostate cancer. Results. Regarding the patients, 31.9% were African American and 66% had pathologic Stages C or D1 disease. Only 9.6% of patients with Stages A2 and B had a prostate-specific antigen (PSA) value greater than 10 ng/ml, whereas 97% of patients with PSA values greater than 20 ng/ml had pathologic Stages C and D1. PSA levels correlated with Gleason score (P = < 0.05); 51% and 100% of patients with Gleason score 5–7 and 8–10, respectively, had PSA values greater than 10 ng/ml. Twenty-two patients (23%) had DNA aneuploid tumors. Comparisons of mechanical to enzymatic cell suspensions indicated that DNA aneuploidy was better preserved in mechanical cell preparations. DNA ploidy correlated with pathologic stage (P = < 0.05) and Gleason score (P = < 0.05). Fifteen of 79 patients (18.9%) with Gleason score 5–7 had DNA aneuploid tumors versus 71.4% of patients with Gleason score 8–10. PSA groups correlated with ploidy status (P = 0.01). Although the majority of patients (19 of 22) with DNA aneuploid tumors had elevated preoperative PSA levels, none had a PSA value greater than 50 ng/ml. Conclusions. DNA ploidy analysis correlated with established prognostic indicators in prostate cancer; however, its independent correlation with natural history and treatment outcome must be established for it to have an effect on therapeutic decisions.     

大肠癌中CD133、COX-2的表达及DNA倍体的研究

目的 检测大肠癌组织中CD133、COX-2的表达和DNA倍体状态,探讨这些因素与大肠癌临床病理特征的关系及彼此间的相关性。方法 采用免疫组织化学SP法对90例大肠癌组织和90例大肠正常黏膜组织进行CD133、COX-2检测,同时运用流式细胞术对其中30例进行DNA倍体检测。结果 CD133、COX-2的表达及DNA异倍体在大肠癌中显著高于大肠正常黏膜组织(P＜0.05);其中CD133在大肠癌中的表达与组织分化程度及淋巴结转移呈正相关(P＜0.05),COX-2在大肠癌中的表达与TNM分期及淋巴结转移呈正相关(P＜0.05)。此外,CD133、COX-2及DNA倍体在大肠癌中彼此间也呈正相关(P＜0.01)。结论 大肠癌中CD133、COX-2的表达及DNA倍体增加,且彼此间呈正相关性;暗示着这些指标可能与大肠癌的发生、发展密切相关,可能作为临床治疗和研究的重要依据。     

RNA expression differences in prostate tumors and tumor-adjacent stroma between Black and White Americans

Prostate cancer (PCa) in Black Americans (BA) is diagnosed at an earlier median age and a more advanced stage than PCa in White Americans (WA). Tumor-adjacent stroma (TAS) plays a critical role in tumorigenesis of prostate cancer. We examined RNA expression in both tumor and TAS of BA compared to WA. After evaluating the geographical ancestry of each sample, preliminary analysis of our own RNA-seq data of 7 BA and 7 WA TAS revealed 1706 downregulated and 1844 upregulated genes in BA relative to WA PCa patients (p_adj < 0.05). An assessment of published RNA-seq data of clinically matched tumor-enriched tissues from 15 BA and 30 WA patients revealed 932 upregulated and 476 downregulated genes in BA relative to WA (p_adj < 0.05). When TAS and tumor epithelial cohorts were compared for the top 2500 statistically significant genes, immune responses were downregulated in BA vs WA TAS, while T cell-exhaustion pathways and the immune checkpoint gene CTLA4 were upregulated in BA vs WA tumors. We found fewer activated dendritic cells in tumor and more CD8 T-cells in TAS of BA versus WA PCa patients. Further characterization of these differences in the immune response of PCa patients of distinct geographical ancestry could help to improve diagnostics, prognostics, and therapy.     

DNA倍体分析技术在门诊患者子宫颈癌筛查中的作用

目的 探讨DNA倍体分析在门诊患者子宫颈癌筛查中的应用价值.方法 对门诊行子宫颈癌筛查的2 692例女性就诊者同时进行液基薄层细胞学检查、DNA倍体分析,对其中可疑病变者840例进行子宫颈活组织检查.对参与子宫颈癌普查的妇女进行液基薄层制片,分别进行巴氏染色和Feulgen染色,由细胞学医师对巴氏染色片作常规细胞学诊断,应用全自动DNA倍体分析系统对Feulgen染色片进行自动扫描诊断.结果 840例患者子宫颈活组织病理检查结果分别为：慢性子宫颈炎554例（66.0％）,子宫颈上皮内瘤变（CIN）Ⅰ 25例（3.0％）,CINⅡ59例（7.0％）,CINⅢ100例（11.9％）,子宫颈癌102例（12.1％）.DNA倍体分析中可见DNA异倍体细胞者（DNA异倍体细胞阳性）486例,未见DNA异倍体细胞者（DNA异倍体细胞阴性）354例;DNA异倍体细胞阳性、异倍体细胞≥3个用于筛查CINⅡ及以上病变的敏感度、特异度、阳性预测值、阴性预测值分别为91.9％和89.2％、58.5％和35.8％、49.4％和57.3％、94.1％和77.2％;采用液基薄层细胞学检查LSIL及以上用于筛查CINⅡ及以上病变的敏感度、特异度、阳性预测值、阴性预测值分别为40.2％、90.0％、39.6％和76.9％.结论 DNA倍体定量分析技术较常规细胞学敏感性高,可作为子宫颈癌普查的指标之一.     

Reactive stroma component COL6A1 is upregulated in castration-resistant prostate cancer and promotes tumor growth

Castration-resistant prostate cancer (CRPC) remains the most critical challenge in the clinical management of prostate cancer (PCa). Reactive stromal changes in PCa are likely involved in the emergence of CRPC. In the present study, we identified a novel oncogene termed COL6A1 which was upregulated in the reactive stroma of CRPC. We established an androgen-independent LNCaP (LNCaP-AI) cell line in steroid-reduced (SR) medium within 2 months. We examined COL6A1 expression with western blot during the LNCaP-AI induction, and studied the function of COL6A1 in vitro and in vivo. Immunohistochemical staining of COL6A1 was performed in ten pairs of androgen-sensitive PCa and CRPC samples. We demonstrated that COL6A1 expression was markedly increased in LNCaP-AI cells and CRPC tissues compared with LNCaP cells and paired androgen-sensitive PCa specimens. In vitro, COL6A1 knockdown resulted in G1-S cell cycle arrest and descended vitality. Overexpression of COL6A1 was associated with accelerated S phase entry and elevated vitality in prostate cancer cells. COL6A1 also promoted tumorigenesis of LNCaP cells in vivo. Taken together, these data suggest an important role of COL6A1 in the molecular etiology of castration-resistant prostate cancer, and support the potential use of COL6A1 in CRPC therapy.