恶性肿瘤DNA倍体异质性与细胞生物学特性的关系

目的 探讨恶性肿瘤组织DNA倍体异质性与癌细胞生物学特性的关系.方法 采用流式细胞术(FCM)检测163例恶性肿瘤组织DNA倍体异质性,并分析DNA倍体异质性与肿瘤细胞DNA指数(DI)、细胞凋亡(Apo)百分比、S期细胞百分比(SPF)以及DNA倍体类型等细胞生物学特性参数之间的相关性.结果 各种恶性肿瘤的DNA倍体异质性明显不同,在6种肿瘤中,DNA倍体异质体检出率大小分布顺序与DNA异倍体的分布十分相似.恶性肿瘤组织的DI值与DNA异质体检出率呈显著正相关(r=0.872 4,P＜0.05).异质体的DNA异倍体检出率(100.0%)和SPF[(1.51±0.67)%1均显著高于同质体[22.0%,(0.34±0.51)%](P均＜0.01),而Apo百分比[(2.08±1.25)%]显著低于同质体组织[(11.99±10.25)%](P＜0.01).结论 DNA倍体异质性是恶性肿瘤的重要生物学特性之一,且与癌细胞其他生物学特性关系十分密切,可作为肿瘤的恶性程度和预后判断的重要指标.     

恶性肿瘤患者体腔液细胞生物学特性与临床生物学行为的关系

目的探讨恶性肿瘤患者体腔液细胞DNA倍体类型、细胞动力学参数与临床生物学行为之间的关系.方法采用流式细胞术对尚未进行化疗的91例恶性肿瘤患者体腔液细胞DNA含量进行了检测和分析.结果恶性肿瘤患者体腔液细胞DNA异倍体检出率为65.93%.不同体腔液细胞的生物学特性有明显的不同.随着患者临床分期的增加,DNA异倍体检出率、DI值和SPF均逐渐增加;Apo则逐渐减低.肿瘤转移者的异倍体检出率、DI值和SPF均显著高于未转移者(P＜0.05).结论恶性肿瘤患者体腔液细胞生物学特性的区别,反映了肿瘤恶性程度的不同.患者体腔液细胞生物学特性变化与患者临床生物学行为的关系十分密切.因此,体腔液细胞生物学指标,可作为肿瘤进展的监督指标.     

流式细胞术Ki67/DNA双标记法在实体肿瘤中的应用

目的 探讨流式细胞术双标记法检测恶性实体肿瘤细胞DNA含量、细胞周期和Ki67表达和组织学分级之间的关系.方法 利用流式细胞术Ki67/DNA双标记法同时检测87例新鲜恶性实体肿瘤的DNA含量、细胞周期和增殖核抗原Ki67的表达.结果 异倍体发生率为40.2%,其中高分化肿瘤为11.1%,中分化肿瘤为37.5%,低分化肿瘤为46.3%.Ki67阳性细胞为0.5%～87%(17.36%±16.6%).S期细胞百分比为0～24%(5.28%±4.85%).高分化肿瘤S期细胞百分比及增殖核抗原Ki67的表达明显低于中分化和低分化肿瘤.统计学上有显著性差异(P＜0.01).中分化和低分化肿瘤之间则差异无显著性(P＞0.05).异倍体肿瘤S期细胞百分比高于二倍体肿瘤,差异有显著性(P＜0.01),而Ki67的表达在两者之间无显著性差异.结论 流式细胞术Ki67/DNA双标记法可同时检测实体恶性肿瘤细胞的DNA含量、细胞周期和增殖核抗原Ki67的表达,并能进一步阐明这些参数与组织学分级的关系.方法 简便、快速,有利于对肿瘤生物学特性的了解.     

DNA倍体分类与癌组织生物学特性的关系

目的　探讨恶性肿瘤DNA倍体分类的某些生物学特性的依据。方法　采用流式细胞术对 1 0 0例恶性肿瘤的癌组织和癌旁组织进行了DNA含量分析 ,并对癌组织不同倍体类型与某些生物学特性的关系进行了探讨。结果　把癌组织DNA倍体类型分为 :二倍体 (D)、近二倍体 (ND)、四倍体 (T)、非整倍体 (AN)和多异倍体 (M)。后四种倍体类型统称为DNA异倍体 (H)。癌组织DNA类型不同 ,其增殖活性明显不同。从D→ND→T→AN→M ,其SPF和PI逐渐升高 ;在DNA异倍体肿瘤患者中 ,其癌旁组织DNA异倍体检出率也逐渐升高 ;另外DNA异倍体患者癌旁组织异倍体检出率显著高于二倍体者 (P <0 0 5)。结论　癌组织DNA倍体类型从D→ND→T→AN→M ,其SPF、PI和癌旁组织DNA异倍体的检出率均逐渐升高。这表明从D→ND→T→AN→M ,肿瘤细胞恶性度愈来愈高。这为癌组织DNA倍体分类提供了生物学依据。     

DNA倍体分类与癌组织生物学特性的关系

目的：探讨恶性肿瘤DNA倍体分类的某些生物学特性的依据。方法：采用流式细胞术对100例恶性肿瘤的癌组织和癌旁组织进行了DNA含量分析，并对癌组织不同倍体类型与某些生物学特性的关系进行了探讨。结果：把癌组织DNA倍体类型分为；二倍体（D）、近二倍体（ND）、四倍体（T）、非整倍体（AN）和多异倍体（M）。后四种倍体类型统称为DNA异倍体（H）。癌组织DNA类型不同，其增殖活性明显不同。从D→ND→T→AN→M,其SPF和PI逐渐升高；在DNA异倍体肿瘤患者中，其癌旁组织DNA异倍体检出率也逐渐升高；另外DNA异倍体患者癌旁组织异倍体检出率显著高于二倍体者（P＜0．05）。结论：癌组织DNA倍体类型从D→ND→T→AN→M，其SPF、PI和癌旁组织DNA异倍体的检出率均逐渐升高。这表明从D→ND→T→AN→M，肿瘤细胞恶性度愈来愈高。这为癌组织DNA倍体分类提供了生物学依据。     

胃癌细胞增殖与P-gp表达的相关性研究

目的 了解胃癌细胞增殖和P-糖蛋白（P-gp）表达水平的临床意义。方法 采用流式细胞术（FCM）检测95例胃癌手术标本P-gp、DNA指数（DI）、S期细胞百分率（SPF）和细胞增殖指数（PI）,结合临床病理资料进行相关性分析。结果 胃癌P-gp表达阴性39例（41.1%）;P-gp表达阳性有56例（58.9%）,其中轻、中度表达有29例（30.5%）,重度表达有27例（28.4%）。95例胃癌中非整倍体肿瘤48例（50.5%）,非整倍体肿瘤SPF和PI显著高于二倍体肿瘤（P〈0.01）,非整倍体肿瘤P-gp表达低于二倍体肿瘤,但差异无统计学意义。胃癌低分化组与中分化组的肿瘤倍体例数比较有非常显著性差异（P〈0.01）;胃癌中-低分化组与中分化组的肿瘤倍体例数比较有非常显著性差异（P〈0.01）。以胃癌发生部位、肿瘤大小、临床分期和有无淋巴结转移进行分组比较SPF、PI、P-gp和倍体关系,均无显著性差异。结论 用FCM检测胃癌P-gp表达水平和DNA倍体可为临床选择适当的化疗方案提供参考依据。     

大肠癌细胞增殖动力学与临床病理学研究

目的 应用流式细胞术(FCM)对64例大肠癌的石蜡包埋组织的DNA指数(DI)、S期细胞百分比(SPF)、细胞增殖指数(PI)进行检测,并且对上述三项指标与大肠癌的组织类型、组织分化程度、侵犯深度、转移等临床病理因素的关系进行探讨。方法 应用流式细胞仪FACS Calibur,激发波长为488nm,DNA染色试剂为PI,检测上述参数,分析采用Modfit LT2.0软件。结果 在癌组织分化低的大肠癌组中,上述三项指标均高于高—中分化腺癌组,且具有显著意义(P<0.05,P<0.01)。粘液腺癌的SPF、PI值均高于单纯腺癌组(P<0.01)。在有淋巴结转移的大肠癌中DI增高(P<0.05)。在有血行转移的大肠癌中DI及SPF、PI增高(P<0.05,P<0.01)。结论 用石蜡包埋组织进行DNA倍体及细胞周期分析的回顾性研究,能反应肿瘤的生物学特性,参数DI、SPF、PI能够为大肠癌细胞群体的增殖活性与病理因素之间相关性的研究提供依据。     

非霍奇金淋巴瘤流式细胞术免疫表型检测及DNA倍体分析

 目的 探讨非霍奇金淋巴瘤（NHL）流式细胞术免疫表型检测及DNA倍体分析与疾病的诊断、分型、恶性程度判断及预后之间的关系。方法 对每例NHL患者进行病理形态学及免疫组化分类,标本均采用淋巴结活检或细针穿刺获取新鲜淋巴结组织,用单克隆抗体标记及碘化丙啶染液一步插入性DNA定量染色法染色后行多参数流式细胞学检测及DNA倍体（DI,SPF）分析。结果 淋巴结常规病理中3例B细胞肿瘤和1例T细胞肿瘤未能明确分型,经流式检测诊断明确。NHL组细胞DI值,DNA异倍体检出率和SPF高于正常对照组,差异有统计学意义（P＜0.01）。DNA四倍体、非整倍体和多异倍体细胞SPF、DI值显著高于二倍体细胞（P＜0.05）,在DNA异倍体细胞中,从DNA异倍体细胞ND→T→ AN →M,SPF检出率越来越高。NHL不同病理分组和NHL-Ⅲ、NHL-Ⅳ组与对照组间异倍体检出率、SPF值及DI值,差异均有统计学意义（P＜0.05）。结论 免疫表型单克隆抗体在不同类型淋巴瘤有不同的表达,可提高淋巴瘤诊断分型的准确率;DI,SPF水平可反映肿瘤增生情况及病理学恶性程度且与预后有关。     

恶性肿瘤复发患者血细胞生物学特性研究

目的 :探讨肿瘤复发时患者血细胞生物学特性的变化规律。方法 :用流式细胞术对 6 0例恶性肿瘤复发患者外周血细胞的DNA倍体、凋亡水平和增殖活性进行了检测。结果 :在 6 0例肿瘤复发患者血细胞中有 4例出现了DNA异倍体细胞 ,检出率为 6 6 7% ,其DNA异倍体DI值为 1 2 5± 0 2 2 ,其异倍体类型 3例为非整倍体 ,1例为近二倍体。肿瘤复发者血细胞的凋亡水平和增殖活性均显著高于未复发者 (P <0 0 5 )。结论 :肿瘤复发可导致患者血细胞出现某些细胞生物学特性的变化。这种变化可作为确定肿瘤复发的生物学参考指标 ,也为肿瘤复发规律研究积累了生物学资料。     

化疗对非小细胞肺癌PCNA表达及肿瘤细胞DNA含量的影响

目的研究化疗对非小细胞肺癌PCNNA表达和肿瘤细胞DNA含量的影响,及PCNA阳性指数与DNA含量的关系.方法用免疫组化S-P法检测两配对组各18例非小细胞肺癌PCNA表达,并用图像分析系统进行肿瘤DNA分析、倍体测量.结果化疗后组与对照组肿瘤在PCNA阳性指数(PI)、DNA指数、平均DDNA质量、S期比率及＞5C细胞百分比均有显著性差异(P＜0.05,P＜0.01).且PI与DNA含量之间呈正相关关系.但两组间异倍体肿瘤率无显著差异(P＞0.05).结论联合化疗影响肿瘤细胞PCNA表达及DNA含量,即降低了肿瘤的增殖活性.     

DNA ploidy and cell-cycle analysis in intracranial meningiomas and hemangiopericytomas: A study with high-resolution DNA flow cytometry

Although various DNA flow-cytometric studies have been performed on meningiomas, the role of DNA ploidy and the S-phase fraction (SPF) in predicting biological tumor behavior remains unresolved. Discrepant results in earlier studies might be due to different preparing, staining and measuring techniques; different quality standards; and lack of sophisticated computer software. In this study, high-resolution DNA flow cytometry using the DNA-specific dye DAPI (4′, 6′-diamidino-2-phenylindol) was performed on stored frozen tissue from 128 microsurgically resected meningiomas and 7 hemangiopericytomas, including 17 recurrent meningiomas and 4 recurrent hemangiopericytomas. The computer software Multicycle 2.5 was used to determine the ploidy level and to perform cell-cycle analysis. DNA aneuploidy and SPF were significantly higher in atypical, anaplastic and recurrent meningiomas and correlated well with histopathological features such as focal necrosis, infiltration of dura mater and mitotic activity. Among 128 meningiomas, 42 had additional DNA aneuploid stem lines. No association between hypo- and hyperploidy and either histological subtype or clinical outcome was found. In 7 hemangiopericytomas, SPF was significantly higher compared to the benign meningioma group, while only 1 tumor was aneuploid. In all 42 DNA aneuploid tumors, cell-cycle analysis was performed separately for the euploid and aneuploid stem lines. The proliferation parameters (SPF, G₂/M phase) were significantly higher in the DNA aneuploid stem lines. DNA ploidy and SPF are thus useful indicators of different biological behavior within identical histological subgroups in meningiomas. Int. J. Cancer (Pred. Oncol.) 79:116–120, 1998.© 1998 Wiley-Liss, Inc.     

石蜡包埋组织DNA含量的流式细胞分析对恶性淋巴瘤的诊断价值

 目的 探讨流式细胞术（FCM）对恶性淋巴瘤的DNA含量分析在肿瘤诊断、临床分期、分类中的意义。方法 采用流式细胞仪检测48例恶性淋巴瘤（ML）和30例淋巴结反应性增生（RLN）,石蜡包埋组织,并分析它们的DNA倍体及细胞周期变化。结果 48例恶性淋巴瘤石蜡包埋组织样本中17例为异二倍体,31例为二倍体,30例RLN为二倍体。恶性淋巴瘤的S期细胞比率（SPF）、增生指数（PI）均高于淋巴结反应性增生（P＜0.05）。结论 异倍体的出现对恶性淋巴瘤的诊断具有较高特异性。SPF,PI对恶性淋巴瘤和淋巴结反应性增生的鉴别具有重要意义。     

食管鳞状细胞癌DNA含量和倍体分析的临床意义

目的:研究食管鳞状细胞癌DNA含量的分布及倍体类型与其生物学行为的关系。方法:采用流式细胞技术对手术切除的106例食管鳞癌患者新鲜标本进行DNA分析,分析DNA含量、S期细胞分数、增殖指数及倍体类型与食管鳞癌浸润深度、淋巴结转移、病理分级及分期等生物学行为的关系。结果:食管鳞癌患者异倍体出现率为82.08%。DNA含量、S期细胞分数、增殖指数随TNM分期增加呈现逐渐增高的趋势,但与肿瘤浸润深度(T分期),病理分级不相关。有淋巴结转移组食管鳞癌S期细胞分数和增殖指数均高于无淋巴结转移组,DNA含量两组间差异并无统计学意义。异倍体食管鳞癌DNA含量、S期细胞分数、增殖指数及淋巴结转移均明显高于二倍体食管鳞癌。结论:食管鳞癌DNA含量的分布及倍体类型与其生物学行为相关,DNA分析是评估食管鳞癌预后、制定术后治疗方案的一个有价值的参数。     

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.     

胃癌细胞DNA倍性,增殖活性与预后的关系

用液式细胞术（ＦＣＭ）测定７４倒胃癌细胞ＤＮＡ倍性，Ｓ期细胞比率（ＳＰＦ）和增殖指数（ＰＩ），以１８例正常胃粘膜作为正常对照。结果：胃癌组６６．２３％（４９／７４）检出异倍体，正常组均为二倍体，两者差异显著（Ｐ＜０．０１）．而胃癌组ＳＰＦ和ＰＩ也均显著高于对照组（Ｐ＜０．０１）．ＳＰＦ，ＰＩ与胃癌淋巴结转移、切端癌残留有关（Ｐ＜０．０１，Ｐ＜０．０５）．异倍体胃癌患者五年生存率显著低于二倍体胃癌（Ｐ＜０．０５）．年龄＞５５岁组的异倍体胃癌预后更差（Ｐ＜０．０５）．ＳＰＦ，ＰＩ与胃癌术后生存呈显著负相关（Ｐ＜０．０１）．ＦＣＭ胃癌细胞ＤＮＡ倍性、增殖活性检测有助于胃癌恶性度和预后的判断。     

S-phase fraction and DNA ploidy in 633 T1T2 breast cancers: a standardized flow cytometric study.

The lack of a standardized methodology for quantifying DNA ploidy and S-phase fraction (SPF) by flow cytometry is hindering routine use of these markers in breast cancer management. In a retrospective clinical multicenter study, we validated a standardized flow cytometry protocol. We tested 633 frozen T(1)T(2), N(0)N(1), M(0) breast tumors obtained in four institutions. Cell preparation was standardized, and precise rules for data interpretation were followed. Three SPF classes were defined on the basis of tertiles after adjustment for ploidy. DNA aneuploidy was observed in 61.0% of cases. No significant difference was observed among centers. Aneuploidy and high SPF were associated with large tumor size, node involvement, high histological grade, and hormone receptor negativity. In the overall population (median follow-up, 69 months), patients with medium and high SPF values had shorter disease-free survival (DFS) than those with low SPF values (P < 0.0001). Ploidy had no significant influence. By Cox analysis, SPF, pN, and estrogen receptor status were independent predictors of DFS (P = 0.0002, P = 0.001, and P = 0.05). In node-negative patients, SPF was the only predictor of DFS (P = 0.01), whereas in node-positive patients, the risk of relapse increased with both high SPF (P = 0.003) and estrogen receptor negativity (P = 0.004). Low SPF values distinguished grade II tumors with a particularly good outcome. Our results strongly support the use of SPF in multicenter studies and clinical trials and suggest that node-negative patients with slowly proliferating tumors do not require systemic adjuvant therapy.     

恶性肿瘤组织DNA倍体分类与细胞动力学的关系

目的：探讨恶性肿瘤患者肿瘤组织细胞DNA倍体分类与细胞动力学的关系。方法：采用流式细胞术对2504例恶性肿瘤组织DNA含量进行了检测，对DNA倍体进行了分类，并分析了DNA倍体类型与细胞凋亡（Apo)检出率及S期细胞比率（SPF）的关系。结果：恶性肿瘤组织DNA倍体分型：二倍体（D，473例）、近二倍体（ND，49例）、四倍体（T，307例）、非整倍体（AN，1040例）和多异倍体（M，635例）。后4种统称为DNA异倍体（H）共2031例。恶性肿瘤组织Apo检出率显著低于正常对照组（Ｐ＜０．０１），而SPF比率却显著高于正常对照组（P＜0．01）。二倍体肿瘤组织Apo检出率显著高于异倍体肿瘤组织（P＜0．01），SPF比率显著低于异倍体组织（P＜0．01）。在恶性肿瘤组织中，含有不同数量异倍体克隆者，不同DNA倍体异质性质的Apo检出率和SPF比率均有显著性差异（P＜0．01）。不同DI值的肿瘤组织其Apo检出率和SPF比率有明显不同。结论：恶性肿瘤能导致肿瘤组织Apo水平显著降低，细胞增殖活性显著增加。而且这些变化与肿瘤组织倍体类型、DI值、DNA异倍体克隆数以及DNA倍体异质性等均有十分密切的关系。     

Prognostic value of S-phase fraction and DNA ploidy studied with in vivo administration of bromodeoxyuridine on human gastric cancers

The authors studied the prognostic values of DNA ploidy pattern and proliferative activity with in vivo administration of bromodeoxyuridine in human gastric cancers. Fresh specimens surgically removed from 117 patients with gastric cancer were investigated by flow cytometric study using a monoclonal antibody to bromodeoxyuridine. DNA ploidy patterns were classified into four types according to the bivariate BrdUrd/DNA distribution: D1, tumors with single diploid population; D2, tumors which showed mosaic of diploid and aneuploid population; A1, tumors with single aneuploid population; and A2, several aneuploid populations without diploid population. The numbers of cases of each ploidy pattern were as follows: D1, 36 cases (30.8%); D2, 38 cases (32.5%); A1, 15 cases (12.8%); and A2, 27 cases (23.1%). DNA ploidy pattern and S-phase fraction (SPF) showed no relation with clinicopathologic findings, except for type A2. In type A2, lymph node metastasis and lymphatic vessel invasion were observed more often than type D1. The SPF calculated from the bivariate BrdUrd/DNA distribution was higher in aneuploidy (D2, A1, and A2) than in diploidy (D1) (P less than 0.01). Also, A2 exhibited a higher SPF than A1 (P less than 0.01). Furthermore, SPF correlated with DNA index significantly (P less than 0.01). Patients who showed aneuploid tumors, DNA ploidy type A2, or SPF of more than 10% survived 3 years less than those with diploid tumors, DNA ploidy type D1, or SPF of less than 10%, respectively (P less than 0.05). By analyzing with the Cox's proportional hazard's model, it is revealed that DNA ploidy and SPF are one of the independent factors of prognostic significance. The results indicated that the patients with aneuploid tumors or highly proliferative tumors had a poor prognosis and that DNA ploidy pattern and SPF were useful prognostic factors for gastric cancers.     

局部晚期宫颈鳞癌流式细胞术分析与放射治疗预后的关系

 目的 研究局部晚期宫颈鳞癌流式细胞术分析结果（DNA倍体、SPF和PI）与放疗预后的关系，探讨它们在预测宫颈癌放疗效果中的价值。方法 68例ⅢB期宫颈鳞癌病人在放疗前钳取宫颈癌组织，制备成单细胞悬液，采用流式细胞术检测癌细胞DNA倍体、S期比例（SPF）及增殖指数（PI），分析他们及其他临床参数与患者根治性放疗后疾病复发和生存的关系。结果 在68例标本中，异倍体检出率为47．1％（32／68），SPF和PI值为分别为（7．49±2．91）和（12．89±3．75）；在随访期间，全组宫颈癌患者复发率为44．1％（30／68）；二倍体组和异倍体组的5年无复发生存率分别为52．7％和35．2％（P〈0．（15），高SPF组和低SPF组5年无复发生存率分别为25．3％和56．6％（P〈0．05），高PI组和低PI组5年无复发生存率分别为37．5％和49．3％（P〈0．05）；经多因素分析显示，肿瘤大小和SPF是影响宫颈癌患者无复发生存的独立预后因素。结论 通过流式细胞术检测宫颈癌组织SPF可以预测局部晚期宫颈鳞癌放射治疗后的预后。