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。     

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.     

NHL患者骨髓细胞DNA倍体类型与细胞动力学参数的关系

目的：探讨NHL急者骨髓细胞DNA倍体类型与细胞动力学参数之间的关系。方法：采用流式细胞术对尚未进行化疗的327例NHL患者骨髓细胞DNA含量进行了检测，并分析DNA倍体类型与细胞凋亡(Apo)水平及S—期细胞比率(SPF)的关系。结果：NHL患者骨髓细胞DNA倍体类型分为二倍体(D，136例)，异倍体(H，191例)。患者骨髓细胞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NHL患者骨髓Apo检出率显著高于并倍体肿癌组织(P＜0．01)，SPF比率显著低于异倍体组织(P＜0．01)。在NHL患者骨髓细胞中，含有不同数量并倍体克隆者的Apo检出率和SPF比率均有显著性差异(P＜0．01)。SPF升高者的骨髓细胞的Apo和DNA并倍体检出率均显著低于SPF降低者(P＜0．05)。结论：NHL能导致患者骨髓细胞Apo水平显著降低，细胞增生活性显著增加。而且这些变化与肿癌组织倍体类型和DNA异倍体克隆数均有十分密切的关系。     

人体正常组织的流式细胞术DNA含量参数检测

[目的]建立人体各种正常组织的流式细胞术细胞动力学参数。[方法]用流式细胞术检测337例恶性肿瘤或良性疾病患者某些正常组织的DNA指数(DI)、DNA倍体类型、细胞凋亡水平和增殖活性。[结果]在337例患者的正常组织中DI值为1.00±0.00,DNA倍体为2倍体 ,其细胞凋亡水平为2.67%±2.50% ,S期细胞比率为5.00%±4.59 %。在不同类组织中 ,某些组织的SPF检出率有显著性差异(P<0.05或P<0.01)。恶性肿瘤患者正常组织的Apo和SPF均显著高于良性疾病患者的正常组织(P<0.01和P<0.05)。[结论]在肿瘤研究中选择正常对照组织时 ,恶性肿瘤患者的“正常”组织未必能真正代表机体的正常组织 ;而选择良性疾病患者的正常组织作为肿瘤研究的正常对照组织 ,这种组织可能更接近无任何疾病的正常机体组织的生物学特性。     

卵巢癌组织DNA含量和细胞增殖水平测定及临床意义

[目的]探讨卵巢癌组织的DNA含量(DI)和细胞增殖水平(SPF)与临床病理特征及预后的关系.[方法]应用流式细胞术检测45例卵巢癌患者的新鲜癌组织中DI及SPF.[结果]45例患者中,22例为异倍体.DI为0.90～3.56,中位1.10±0.20.SPF中位11.4%±2.3%.SPF＜5%为23.9%,SPF 5%～14.5%为38.4%,SPF值＞14.5%为37.7%.晚期、透明细胞癌患者DNA异倍体率及DI与SPF均高于早期及其它病理类型患者,但无统计学差异;卵巢癌FIGO分期、病理类型及血清CA125水平与DNA倍体、DI、SPF无关;DNA倍体、DI及SPF与卵巢癌病理分级具有显著相关性.单因素COX分析显示,DNA倍体、DI与卵巢患者的生存期无显著相关性.SPF与生存期显著相关(P=0.009).[结论]应用流式细胞术检测卵巢癌DNA含量及SPF能为高SPF及低分化、透明细胞癌的卵巢癌患者的预后判断提供重要信息.     

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。     

食管癌倍体异质性分析的临床意义

作者采用流式细胞测量术（ＦＣＭ）对３０例食管癌患者１５０份样品进行了ＤＮＡ倍体异质性分析。结果表明，患者癌中心灶ＤＮＡ倍性异质体的检出率为５３．３％。ＤＮＡ倍性异质体和同质体患者癌中心灶和癌旁粘膜组织细胞的生物学特性均明显不同。患者癌中心灶倍性异质体的检出率随癌组织分级、肿瘤体积和癌细胞浸润深度的增加而升高（ｒ＞ｒ０．０５）。ＤＮＡ倍性异质体患者的存活时间显著短于同质体患者（Ｐ＜０．０５）；而异质体患者的死亡率却显著高于同质体患者（Ｐ＜０．０５）。     

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。     

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。     

恶性肿瘤患者体腔液细胞生物学特性分析

[目的]探讨恶性肿瘤患者体腔液细胞DNA倍体类型与细胞动力学参数之间的关系。[方法]采用流式细胞术对未进行化疗的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]二倍体体腔液细胞的凋亡细胞检出率显著高于异倍体者(P<0.01),S鄄细胞比率(SPF)则与此相反(P<0.05)。在异倍体患者中,随体腔液异倍体细胞百分率的升高,凋亡细胞百分率与之呈显著的负相关(r=-0.8086,P<0.05),SPF则呈显著性正相关(r=0.9091,P<0.05)。不同数量异倍体克隆者的凋亡细胞检出率和SPF比率均有显著性差异(P<0.01)。SPF增高者的体腔液细胞的凋亡细胞和DNA异倍体检出率均低于SPF降低者(P<0.05)。[结论]恶性肿瘤能导致患者体腔液细胞的细胞凋亡水平显著降低,细胞增殖活性显著增加。而这些变化与体腔液细胞倍体类型、DNA异倍体细胞检出率和DNA异倍体克隆数均有十分密切的关系。     

鼻咽癌新鲜肿瘤组织DNA倍体性与预后的关系

背景与目的:因肿瘤有生物学异质性,部分肿瘤的预后和TNM分期并不符合;寻找有效的生物学预后指标作为临床分期的补充,可为今后鼻咽癌的个体化治疗提供一个新的依据.本研究探讨初治鼻咽癌患者新鲜肿瘤组织细胞的DNA倍体性与疗效、预后的关系.方法:1999年1月至2000年2月,53例初治鼻咽癌患者进入本研究,其中单纯放疗32例,另21例患者于放疗第4周接受了一个疗程的PF方案化疗.患者治疗前均活检取新鲜肿瘤组织,用流式细胞仪进行DNA倍体检测.结果:53例患者中,二倍体32例(60.4%),异倍体21例(39.6%).不同倍体组患者的年龄、性别、临床分期、N分期、化疗与否的差异无统计学意义(P=0.695、0.657、0.088、0.972和0.335).全组患者中位随访时间73个月(12～84个月).全组5年总生存率65.61%,其中二倍体组为80.92%,异倍体组为42.86%(P=0.002);5年无远处转移生存率二倍体组为84.26%,异倍体组为44.53%(P=0.003);5年无复发生存率二倍体组为92.59%,异倍体组为72.65%(P=0.118).单因素分析结果显示,临床分期是无复发生存率的影响因素,DNA倍体性、临床分期和T分期是总生存率和无远处转移生存率的影响因素.多因素分析结果显示,与总生存率相关的独立预后因素为DNA倍体性(P=0.020)和临床分期(P=0.007),与无转移生存率相关的预后因素亦为DNA倍体性(P=0.017)和临床分期(P=0.011).结论:采用流式细胞术检测新鲜组织细胞的DNA倍体性和临床分期一样可以预测鼻咽癌患者的预后;DNA异倍体患者比二倍体患者更容易出现远处转移而导致治疗失败.     

肿瘤转移患者外周血细胞的某些生物学特征

目的:探讨肿瘤转移患者外周血细胞的生物学变化特征。方法:用流式细胞术对396例肿瘤转移患者和371例非转移患者外周血细胞的DNA倍体、凋亡水平和增殖活性进行了对比分析。结果:在有肿瘤转移的396例患者外周血细胞中,有45例出现了DNA异倍体,其检出率为11.36%;DNA异倍体患者血细胞的DNA指数(DNAindexDI)值为1.25±0.26。另371例非转移患者未发现DNA异倍体细胞。肿瘤转移者的细胞凋亡率和S期细胞比率均显著高于非转移者P<0.05,多器官转移者的细胞凋亡率和S期细胞比率也显著高于单器官转移者P<0.05。结论:肿瘤转移能引起患者外周血细胞中出现DNA异倍体,并使血细胞凋亡水平和增殖活性上调。     

Prognostic significance of flow cytometric analysis of DNA content in colorectal cancer: A prospective study

We prospectively analyzed the tumor DNA content by flow cytometry in 100 patients who underwent a curative resection for colorectal cancer between August 1989 and May 1992 in order to evaluate the prognostic significance of DNA ploidy and the DNA index (DI). Patients with aneuploid tumors were found to have a significantly shorter disease-free survival than those with diploid tumors (P = 0.014). In addition, patients who had tumors with a DI greater than 1.6 had a significantly shorter disease-free survival than those who had tumors with a DI of less than 1.6 (P = 0.0001). After stratification by stage, this association was only seen in Dukes' stage C disease (P = 0.0065). Cox's regression analysis demonstrated that the DI (below or above 1.6) rather than DNA ploidy was an important independent predictor of disease-free survival. These results suggest that the DI rather than DNA ploidy provides us with important prognostic information in patients undergoing curative surgery for colorectal cancer. © 1993 Wiley-Liss, Inc.     

Cytofluorometric DNA ploidy analysis in giant cell tumor of bone: histologic and prognostic value.

DNA ploidy analysis by DNA cytofluorometry was performed on 41 tumors obtained from 37 patients with primary giant cell tumor of bone (GCT). Histologically, 26 of the tumors from primary or recurrent lesions were evaluated as grade I, and 13 tumors as grade II. Among the 33 primary GCT patients, 4 patients had local recurrence or pulmonary metastasis. The DNA ploidy pattern and the percentage of hyperdiploid cells showing a greater DNA content than diploid cells, were obtained from DNA cytofluorometry. All of the 33 primary tumors were diploid. Of 6 recurrent tumors, 4 were diploid and 2 were euploid-polyploid. One of the two pulmonary metastatic tumors was diploid, but another that demonstrated a malignant transformation to malignant fibrous histiocytoma was aneuploid. The percentage of hyperdiploid cells was significantly different between primary and recurrent tumors (P = 0.0188) and between grade I and grade II tumors (P = 0.0052), while there was no difference between primary tumors in the cases that recurred or metastasized and those that did not. Thus, these data indicate that cell proliferative activity is closely correlated with biological aggressiveness and histological grading, although DNA ploidy is not useful for predicting prognosis.     

Flow cytometric analysis of tumor DNA profile related to response to treatment and survival in small-cell lung cancer

Flow cytometric (FCM) analysis of tumor DNA ploidy and S-phase fraction (SPF) has been widely used to predict prognosis and treatment response in many malignant tumors, but rarely in small-cell lung cancer (SCLC). In the present study, tumor DNA ploidy and SPF were measured from paraffin-embedded tumor biopsy samples of 36 small-cell lung cancer patients treated with combination chemotherapy and radiotherapy. Aneuploidy was detected in 69% of the tumors. There was a statistically non-significant trend towards more aneuploidy among extensive disease (ED) patients as compared to patients with limited disease (LD): 80% versus 65%, respectively (p = 0.69). The mean SPF was 213% (± 7.6) in patients with LD and 29.0% (± 5.3) in patients with ED, the difference (7.6%) being statistically significant(p = 0.008, 95% CI for the difference 2.2-13.1). No significant differences was detected in the survival of aneuploid and diploid patients or patients with low (⩽24.9%) and high (>24.9%) SPF. Similarly, no significant difference was observed between aneuploid and diploid cases in relation to response to treatment or response duration. It is concluded that the difference detected in the SPF with LD and ED of SCLC may indicate the biological aggressiveness of extensive SCLC.     

DNA ploidy, proliferative capacity and intratumoral heterogeneity in primary and recurrent head and neck squamous cell carcinomas (HNSCC)--potential implications for clinical management and treatment decisions

Despite new diagnostic and therapeutic strategies (combined radiochemotherapy, EGFR antibody Cetuximab), the prognosis of head and neck squamous cell carcinoma (HNSCC) is still poor and more information regarding prognosis is essential to establish earlier and better treatment options. To elucidate the role of DNA ploidy and cellular proliferation, resected tumors of 48 patients with primary or recurrent HNSCC were analyzed by flow cytometry and in vitro-5-bromodeoxyuridine incorporation (BrdU). The results were compared with histopathological findings such as tumor size, lymph node involvement and tumor differentiation. To assess the influence of intratumoral heterogeneity of these biological parameters, multiple biopsies (>3) were analyzed by flow cytometry and BrdU-incorporation in 12 larger (>4 cm diameter) tumors. BrdU-labeling index (LI%) was significantly higher in aneuploid HNSCC and correlated significantly with poor histologic differentiation of the analyzed tumor tissues (P<0.001). Furthermore, a trend for higher LI% in nodal positive tumors was observed. Aneuploid HNSCC showed significantly more often tissue dedifferentiation (P=0.049) and in most cases an advanced tumor stage, especially in tumors with biclonal cell lines. Lymph node involvement was also seen more often in aneuploid and undifferentiated tumors. As in aneuploid tumors recurrent HNSCC showed in most cases a higher LI% and poor tissue differentiation, but as a result of the small collection of samples there was no correlation between aneuploidy and tumor recurrence. To proof the robustness of the acquired data and to estimate the influence of intratumoral heterogeneity to ploidy and LI% multiple biopsies were analyzed in larger tumors. Using a specific statistical algorithm a secure estimation of ploidy and LI% was possible by a single biopsy in these tumors. These findings indicate aneuploidy and proliferative activity as important findings for malignant progression in HNSCC. An estimation of these biological parameters may be useful for identification of patients with high risk for lymph node involvement or tumor recurrence and pre-treatment can be performed by a single biopsy. As a conclusion, these patients may benefit from more aggressive treatment.     

Thymoma. The prognostic significance of flow cytometric DNA analysis.

The clinical course of patients with thymoma varies widely despite its histologically benign appearance. Treatment decisions are based on local invasion and the extent of resection. Because some patients have more aggressive tumors, the prognostic significance of flow cytometric (FCM) analysis of nuclear DNA content was examined. Adequate tissue from paraffin-embedded blocks was available for 25 patients. Using FCM, the percentage of cells in S-phase (%S) and the ploidy, based on the DNA index (DI), were determined. The mean patient age was 52 years, with a female-to-male ratio of 1.3:1 and a median follow-up of 64 months. Seventeen patients underwent total tumor resections, and 12 also received radiation therapy. Eight patients underwent subtotal resections, with five receiving radiation therapy (with or without chemotherapy) and three receiving chemotherapy alone. Based on invasion and intrathoracic dissemination, the tumors were classified into four stages. The mean %S was 5.6. There was no relationship observed between %S and patient outcome. The 5-year disease-free survival rate was 85% for the 16 patients with diploid (DI = 1) tumors and 33% for the 9 patients with aneuploid (DI more than 1) tumors (P less than 0.002). Similar significant differences were observed by stage and extent of surgery. For those who had total resection (n = 17), the disease-free survival rate was 89% when DI equaled 1 and 50% when DI was more than 1 (P = 0.01). Although the numbers studied were small, when stage, histologic findings, and type of surgery were subdivided by DI, a higher incidence of relapse was associated consistently with aneuploidy. The DI appears to be a useful prognostic parameter for identifying patients at high risk of relapse.