Cytofluorometric nuclear DNA analysis and immunohistochemical study on the proliferative activity of the trophoblasts in human early gestation

Changes in proliferative activity in trophoblastic cells were evaluated in various different sites of implantation. DNA content analysis by cytofluorometry and immunohistochemical study using a monoclonal antibody Ki-67 was performed in 5 cases with a gestation period of 8-9 weeks. The measurement of proliferative activity of trophoblastic cells was analysed in four different sites of implantation: (1) the top of the anchoring villi, (2) the cell column, (3) the intermediate trophoblasts in the decidua basalis and myometrium, and (4) the free villi. A large number of Langhans cells on the top of the anchoring villi (L-cells) were mononuclear and tetraploid. It was demonstrated that the L-cell group had 2C, 4C and 8C peaks and also had a 4C mode. S-phase fraction was also found in the L-cell group. The so-called intermediate trophoblasts (IMT) in the cell column, decidua basalis and myometrium had 2C and 4C peaks and no S-phase fraction. Ki-67 positive cells existed only in the L-cell group. The findings are as follows: 1. The L-cell group had a higher proliferative activity than the other trophoblasts. It was found to have a mononuclear tetraploidy pattern. 2. It is suspected that the cells having 4C DNA content in the IMT originated in the L-cell group. 3. In conclusion, L-cells may differentiate to syncytial trophoblasts and IMT.     

Evaluation of cellular proliferative activity and DNA ploidy as a prognostic factor in breast cancer

BACKGROUND: In the past years, morphological indexes and kinetic parameters have been introduced to characterize breast cancer in order to select breast cancer patients for adjuvant therapy. The alterations of the proliferative activity of neoplastic cells and DNA ploidy may provide important information about the aggressiveness of the lesion and may be used as powerful prognostic factors. In fact, prognostic information based on the classic clinico-pathologic parameters such as histotype, stage and grade are no longer sufficient to select patients for long term follow-up. The aim of this study was to evaluate the malignant potential of neoplastic cells through the determination of modifications of the proliferative index and the evaluation of DNA ploidy. METHODS: Forty-five breast cancer patients, 32 to 80 years of age, were studied. Proliferative indexes and DNA ploidy, besides the other clinical-pathologic parameters have been evaluated. The proliferative index was assessed with the immunocytochemical determination of Ki67 and quantified with the CAS 200: 31 cases had a high proliferative index, 12 medium and 2 low. DNA ploidy was analyzed on cytologic preparations with static cytometry utilizing an image analyzer CAS 200: 13 cases were diploid, 32 non-diploid. RESULTS: A close relationship was found between Ki67 expression and DNA ploidy. In fact, lesions with a high proliferative index were all non-diploid, whereas, those with a low proliferative index were all diploid. Lesions with a medium proliferative index were 2 diploid and 1 non-diploid. Among the 45 cases, studied in a follow-up period of 36 months, 12 showed disease recurrence; 3 showed a medium Ki67 expression and were diploid, 9 a high Ki67 expression and were all non-diploid. CONCLUSIONS: In conclusion, proliferative indexes and DNA ploidy may be considered additional prognostic parameters which may potentially influence the clinical behaviour of breast cancer and may be utilized besides all the other clinico-pathologic data to assess these lesions.     

A distinct pattern in the DNA ploidy histograms of hydatidiform moles and nonmolar abortuses is caused by accumulation of trophoblasts in the late s-phase.

DNA ploidy analysis is a useful tool to distinguish the partial hydatidiform moles (PMs) from complete hydatidiform moles (CMs) and nonmolar abortuses (NAs). DNA ploidy histograms of hydatidiform moles are sometimes difficult to interpret because of the uneven distribution of nuclei in the S-phase, simulating aneuploid peaks. In this study, we analyzed DNA ploidy histograms of 25 CMs, 16 PMs, and 28 NAs, with special reference to the accumulation of cells in the late S-phase using a high-resolution DNA image cytometry. All the gestational products demonstrated the accumulation of cells in the late part of the S-phase fraction. To objectify the observation, we compared the percentage of cells in the second quarter with that of the third quarter of the S-phase fraction. All the gestational products had significantly lower (P < 0.001) percentage of cells in the second compared with that of the third quarter of the S-phase. The mean ratios of the third quarter to the second quarter in CMs, PMs, and NAs were 1.9, 1.7, and 2.5, respectively. This was significantly different from that of highly proliferative endometrial carcinomas. The knowledge of this specific S-phase fraction distribution in molar and nonmolar pregnancy material is important when interpreting the DNA histograms. The possibility of hypoxia being the cause of this phenomenon is also discussed.     

子宫内膜癌DNA倍体及S期细胞比率测定的临床意义

目的 :研究DNA倍体、S期细胞比率 (SPF)在子宫内膜复合型增生、子宫内膜癌不同临床分期和病理分级中的变化并评价其临床意义。方法 :采用流式细胞术(FCM)测定 39例子宫内膜癌 ,8例复合型增生子宫内膜中DNA倍体和SPF的变化。结果 :DNA异倍体率和SPF在子宫内膜癌中高于复杂型增生子宫内膜 (P <0 0 1) ,晚期、低分化和深肌层浸润子宫内膜癌DNA异倍体率和SPF高于早期、高分化、浅肌层浸润子宫内膜癌。结论 :DNA异倍体率、S期细胞比率在子宫内膜癌的诊断和鉴别诊断、监测病情变化和评价预后方面有重要的临床意义。     

DNA ploidy of human granulosa cells from natural and stimulated in vitro fertilization cycles

OBJECTIVE: To analyze and compare the DNA ploidy of granulosa cells from natural and gonadotropin-stimulated follicles obtained during IVF. DESIGN: Retrospective analysis of laboratory data. SETTING: University medical center. PATIENT(S): Seventy-three aspirates of dominant follicles from natural IVF cycles and 113 aspirates from gonadotropin-stimulated cycles were analyzed. INTERVENTION(S): Cytospins were prepared and stained by the Feulgen-thionine method. MAIN OUTCOME MEASURE(S): Image DNA analysis was performed on an automated high-resolution image cytometer. DNA content and the number of nuclei with DNA content >5c were measured. RESULT(S): All samples from natural and gonadotropin-stimulated follicles were found to be diploid. Single cells with DNA content >5c were found in follicular fluid samples of four women with natural IVF cycles and in samples of nine women with gonadotropin-stimulated cycles. CONCLUSION(S): DNA ploidy of granulosa cells from natural follicles has not been studied before. In natural samples, granulosa cells were only diploid, without euploid polyploidization. We were unable to confirm DNA aneuploidy of granulosa cells in gonadotropin-stimulated follicles of women undergoing IVF.     

Cellular angiofibroma of the vulva with DNA ploidy analysis.

Cellular angiofibroma (CAF) is a recently described rare soft tissue neoplasm of the vulva (with only four reported cases) that typically occurs as a well-circumscribed solid rubbery vulvar mass in middle-aged women. The distinct histologic features of bland spindle cells admixed with numerous hyalinized medium to small blood vessels, and a vimentin-positive desmin-negative immunoprofile differentiates this neoplasm from other vulvar tumors such as angiomyofibroblastoma and aggressive angiomyxoma. In this report an additional case of CAF is presented with DNA ploidy analysis and CD99 immunohistochemistry.     

DNA ploidy of ovarian dysgerminomas: correlation with clinical outcome.

Abnormal nuclear DNA content, as determined by flow cytometry, when combined with conventional prognostic variables such as tumor grade or stage at diagnosis, appears to identify patients who are at increased risk for recurrence of disease. The DNA content of ovarian dysgerminoma, a tumor that is homologous to testicular seminoma and is found in young women of childbearing age, was studied to determine if there is a correlation between DNA content and outcome. Such information would be useful in selecting treatment regimens and making possible the preservation of childbearing potential in women who are likely to have a good outcome. The specimens from 23 cases of ovarian dysgerminoma seen at our institution between 1950 and 1985 were analyzed by DNA flow cytometry. Five of the tumors were diploid (21%) and nineteen were nondiploid (79%). Patient outcome was not predicted any better by nuclear DNA content than by conventional prognostic variables.     

Flow cytometry of normal, hyperplastic, and malignant human endometrium. A study of ploidy and proliferative indices including comparison with in vitro S-phase labeling

The validity of 48 endometrial proliferative indices derived from flow cytometric deoxyribonucleic acid histograms was tested by comparison with S-phase labeling indices. There was a positive but weak correlation. Despite this correlation, there was a good relationship between the flow cytometric proliferative indices and both the menstrual phase and degree of tissue differentiation in a larger series of 162 normal, hyperplastic, and malignant endometrial specimens examined by flow cytometry alone. Flow cytometric proliferative indices may be useful for prediction of tumor behavior in non-DNA aneuploid endometrial carcinomas that are assessed as being a good prognostic type by other tumor parameters.     

人胎盘滋养细胞分选技术研究进展

人胎盘滋养细胞(TB)包括绒毛细胞滋养细胞(VCT)、合体滋养细胞(VST)和绒毛外细胞滋养细胞(EVT)。各种TB形态差异较大,在母-胎界面的功能也各不相同,此外由于母体细胞的污染,因此有必要对TB的类别加以区分。如何在保证细胞活性的同时提高纯度成为构建TB细胞系的主要困难之一。梯度离心、磁珠分选和流式细胞术是广泛应用的细胞分选技术。各种分选技术在TB研究中各有利弊,磁珠分选结合流式细胞术是目前较为有效的分选策略。     

Trophic effects of first-trimester human trophoblasts and human chorionic gonadotropin on lymphocyte proliferation

We examined the effects of pure first-trimester human trophoblast cells grown in long-term cultures or their secreted products on the proliferation of human peripheral blood lymphocytes cultured alone, under allogeneic stimulation, or in the presence of concanavalin A. Both trophoblasts and their culture supernatants stimulated lymphocyte proliferation. Culture supernatant had a moderate enhancing effect on lymphocyte mitogenesis in mixed lymphocyte cultures and in the presence of concanavalin A. Anti-human chorionic gonadotropin antibody suppressed the proliferative effect of trophoblast cells and their supernatants in the above experiments in a dose-dependent manner. At physiologic concentrations, both pure and impure forms of human chorionic gonadotropin enhanced (in a dose-dependent manner) proliferation of lymphocytes cultured alone (peak stimulation index, 6 to 7.8 at approximately 7 IU/ml). At higher concentrations (20 to 400 IU/ml) the proliferative effect was abolished. Trophoblast culture supernatant induced the expression of interleukin-2 receptors on lymphocytes after 48 hours of incubation. The supernatant also stimulated proliferation of human colon carcinoma cells. Thus trophoblast or trophoblast-derived human chorionic gonadotropin has a lymphocytotrophic function that may have implications for fetal survival.     

人乳头状瘤病毒感染对孕妇和绒毛滋养细胞的影响

目的 探讨人乳头瘤病毒（ＨＰＶ）感染对孕妇和绒毛滋养细胞铁影响。方法 观察６２例早、中期孕妇尖锐湿疣的病理形态特点,用ＨＰＶ６／１１－ＤＮＡ探针原位杂交法检测其绒毛、胎盘组织及其中１０例外阴、阴道、宫颈尖锐湿疣组织,并用ＨＰＶ６／１１聚合酶链反应（ＰＣＲ）方法验证１０例绒毛及胎盘组织。结果 孕期尖锐湿疣病变发展快,多部位发生比例高,大体形态、组织形态典型者多。ＨＰＶ６／１１－ＤＮＡ原位杂交和ＰＣＲ