Correlation of pretreatment proliferative activity of breast cancer with the response to cytotoxic chemotherapy.

That most cytotoxic agents act specifically against actively proliferating cells is well-recognized. In this study, we attempted to correlate pretreatment S-phase fractions (SPF) measured on DNA histograms with regression of the tumor mass after the administration of neoadjuvant chemotherapy. Tumor cells were obtained from 60 previously untreated, premenopausal patients with no metastases and with noninflammatory disease by fine needle sampling without aspiration. We could evaluate DNA ploidy in all patients and SPF in 50 or 83% of them. Tumor responsiveness was significantly related to SPF. The 12 patients who had SPF of 10% or more showed demonstrable regression; six had complete responses. None of the other parameters tested, i.e., DNA ploidy, histopathologic grade, or hormone receptor content, correlated with response. We believe this information may prove valuable for clinicians as they make their decisions regarding patient therapy.

Enzyme immunoassay of estrogen receptors in fine needle aspirates of breast tumors.

Enzyme immunoassay of estrogen receptors (ER-EIA) was compared to radioligand assay (ER-RLA) in fine needle aspirates of breast tumors. Fine needle aspiration is a relatively atraumatic means of harvesting malignant cells from breast tumors. Fine needle aspiration provides a homogeneous suspension (about 90% malignant cells) with a sufficient amount of cellular material (10 to 50 micrograms DNA per sample) for single point radioligand assays of extractable estrogen (ER) and/or progesterone receptor (PR) in about 85% of primary adenocarcinomas at the time of diagnosis. Sixty-one different samples of malignant mammary cells were obtained by fine needle aspiration from 43 adenocarcinomas, 11 metastatic axillary nodes or 7 cutaneous nodules. Thirteen patients were under antiestrogen therapy (tamoxifen). ER-EIA was performed with Abbott's reagents, following the manufacturer's protocol. ER-RLA was a single saturation (5 nM) dextran-charcoal assay with [3H]R2858 as the labeled estrogen. The sensitivity of ER-EIA allowed dilution of the sample up to 10 times (according to sample cellularity and ER level) with less than 20% deviation from undiluted samples. Three levels of dilution of the samples (1/1, 1/2, and 1/10) allowed them to fall at least once into the range of the ER-EIA standard curve. Quantitative correlation between ER-EIA and ER-RLA was high (r = 0.86), and highest (r = 0.97) when samples from patients undergoing tamoxifen treatment were excluded. Major discrepancies between ER-EIA and ER-RLA appeared in those patients undergoing tamoxifen therapy; much higher values were obtained by ER-EIA. Eight of 13 of these patients were ER negative by ER-RLA but ER positive by ER-EIA. This preliminary observation indicates that in vivo ER modulation by hormones and antihormones should be reevaluated.

Release of estrogen-induced glycoprotein with a molecular weight of 52,000 by breast cancer cells in primary culture.

In an attempt to find estrogen-specific responses in breast cancer, we have established primary cell culture from metastatic pleural effusions of breast cancer and have analyzed the proteins labeled by [35S]methionine and released into the culture medium using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We show that the synthesis of a Mr 52,000 glycoprotein which is released by metastatic breast cancer cells in primary cultures is stimulated by estradiol in four of six patients. This protein is similar to the Mr 52,000 protein of MCF7 cells on the basis of its mobility in one- and two-dimensional gel electrophoresis [the molecular weight of this protein was originally found to be 46,000; it is closer to 52,000 using labeled proteins from New England Nuclear as molecular weight markers], its immunoprecipitation by antisera raised against the Mr 52,000 protein, and its binding to concanavalin A. We conclude that, similar to some breast cancer cell lines, some metastatic breast cancers synthesize a Mr 52,000 glycoprotein which is regulated by estrogens and exported from the cells into the medium. This study also shows that some primary cultures established from metastatic breast cancer remain responsive to estradiol in vitro for the synthesis of specific proteins. More clinical studies are needed to prove the interest of the Mr 52,000 secreted protein as an additional marker of the hormone responsiveness of breast cancer.

Fine-needle cytology of 292 palpable orbital and eyelid tumors.

Fine-needle cytology (FNC) of 292 palpable orbital and eyelid tumors was performed with a 25-gauge (0.5-mm), 3-cm needle and compared with the histopathologic findings in 286 cases. Among these 286 cases, a concordant diagnosis of malignancy and type was achieved in 249 cases (87%). False positive diagnoses were made in four cases (1.6%) and false negative diagnoses in five cases (1.8%). No complications were encountered. These results led to the conclusion that FNC is an accurate tool in the diagnosis of orbital and eyelid tumors, especially when sampling and interpretation are performed by an experienced pathologist.

Prognostic value of the S-phase fraction of breast cancers treated by primary radiotherapy or neoadjuvant chemotherapy.

Neoadjuvant chemotherapy for large operable breast cancers is being increasingly used for the purpose of "downstaging," so that the lesions become accessible to conservative treatment. Since tumor proliferative activity has been shown to be a major prognostic factor for breast cancers, we have studied the value of S-phase fractions established by flow cytometry on cytological samples at diagnosis, in 184 stage II or IIIa patients entered in a randomized trial comparing neoadjuvant to adjuvant chemotherapy. All patients were pre- or perimenopausal, and the median follow-up was for 43 months (24-64). Using the median value (5%) as cutoff, a high SPF was found to be associated with relapse (p < 0.0008), locoregional recurrence (p < 0.02), or metastasis (p < 0.003). However, when the patients were analyzed according to the type of treatment, significance was maintained for the patients in the primary radiotherapy arm (p < 0.003) but not for those in the neoadjuvant chemotherapy arm (p < 0.06). The overall rate of response to neoadjuvant chemotherapy was significantly lower for tumors with low SPF (56.5%), compared to tumors with high SPF (85.6%). Thus, SPF was no longer predictive of outcome when the tumors regressed by more than 50% after chemotherapy (p = 0.66), whereas it was highly predictive in the nonresponding patients (p < 0.0001). Our study has revealed that patients with low-SPF tumors, irrespective of response or treatment schedule, had similar prognosis (around 70% free of disease at 45 months), while the high-SPF nonresponders had a dismal prognosis, with less than 25% free of disease at 24 months. If our results are confirmed with a longer follow-up, proliferative activity of breast cancers should prove to be instrumental for the initial therapeutic decision of stage II or IIIa patients.

Leydig cell tumor of the testis: a case diagnosed by fine-needle sampling without aspiration with histologic, immunohistologic, and electron microscopic analysis.

Fine-needle sampling without aspiration was performed in a patient with a testicular mass. The cytologic diagnosis was consistent with Leydig cell tumor. Cytologic features included abundant grey-blue cytoplasms with spherical or oval nuclei in May-Grünwald-Giemsa-stained smears. Intranuclear inclusions were observed but no Reinke's crystals were detected. Histologic findings confirmed the diagnosis and tumor cells were positive for vimentin. Electron microscopic analysis of the tumor showed abundant smooth endoplasmic reticulum and mitochondria with tubulovesicular cristae but no Reinke's crystals.

Cytologic diagnosis of orbital and periorbital palpable tumors using fine-needle sampling without aspiration.

Sixty-two cases of orbital and periorbital palpable neoplasms were analyzed cytologically. The material was obtained by our technique of simply introducing a fine injection needle in the tumor mass without aspiration. Fifty-six of these tumors had a subsequent histologic diagnosis by surgical procedure or biopsy. Forty-nine of the 56 cytologic diagnoses (87%) were concordant with the histologic findings with regard to malignancy and its variety. In three other cases the diagnosis of malignancy was only achieved by surgical procedure or biopsy (5%). In two cases, there were false-malignant results (4%), one corresponding to a meningioma and the other related to a reactive lymphoid hyperplasia. There was one false-benign (2%) result in a case of non-Hodgkin's lymphoma. In one patient, the cytologic material was insufficient for diagnosis (2%). In six other cases, the initial cytologic examination was ultimately confirmed either by biochemical studies or by biopsies of nodal metastases. No orbital hemorrhage was observed after fine-needle sampling. This outpatient technique is highly accurate and permits diagnosis in a few minutes.

The relation of prognosis to the nuclear diameter of breast cancer cells obtained by cytologic aspiration.

Nuclear diameters of breast cancer cells obtained by aspiration cytology from 245 patients were measured by ocular micrometry. The nuclei of 50 benign lesions were also measured. Cancers with 80% or more of their nuclei larger than 12 mu were classified as "large nuclear type" (LN). Those with nuclei of 12 mu or less were considered "small nuclear type" (SN). One hundred forty-nine cases of the former and 96 of the latter were evaluated. There was significant overlap in nuclear diameter of benign lesions and SN cancers but minimal overlap between SN and LN cancers. Thirty-eight percent of SN cancers were staged as T1 as compared to only 20% of LN tumors. Survival, free of disease, at five years was 90% for the SN cancers and 58% for the LN cancers (p less than 0.001). Intervals between completion of treatment, appearance of metastases and death were longer for patients with SN cancers. Measurement of nuclear diameters of cells obtained by aspiration cytology may, therefore, be useful in determining the prognosis.

[Value and interest of fine needle aspiration cytology in the follow-up of irradiated breast cancers (author's transl)]. / Valeur et intérêt de la ponction cytologique dans la surveillance des cancers mammaires irradiés.

Between 1968 and 1975, the authors studied at the Institut Curie the value of fine needle aspiration cytology in the follow-up of breast cancers, which have been previously treated with definitive irradiation (182 cases) or lumpectomy and post-operative irradiation (20 cases). The cytologic results have been compared to the pathologic aspects of the surgical specimens. The cytologic procedure was made when the tumor decrease was incomplete or too slow, or when, after a free interval, was noted in the irradiated breast a tissue reaction clinically and radiologically indefinite. The post-therapeutic investigation interested 202 cases of breast carcinoma. Compared to the histologic result, the cytologic results proved to be: -- identical. . . . . . . in 170 cases (84%) -- false benign . . . . . in 4 cases (2%) -- false malignant . . . . in 4 cases (2%) -- insufficient cell material in 24 cases (12%) The high rate of correct cytologic diagnosis and the very low rate of wrong results prove the value and interest of the fine needle aspiration cytology for residual tumor masses or for recurrences in irradiated breast cancers.

[Cytology of various rare benign tumors of the breast]. / Cytologie de quelques tumeurs bénignes rares du sein

The authors describe the cytological appearances of three rare benign mammary tumors (granular cell myoblastoma, sebaceous cyst and fibrous histiocytoma), and warn the clinician against over-optimism when presented with a benign cytologic diagnosis. In fact, the proportion of cases in which carcinoma is confused with a benign tumour varies from 3 to 5% according to statistics. They analyse the various causes responsible for the "false benign" diagnosis. The author's opinion is that one can only arrive at the cytological diagnosis of benignity as part of a group of investigations. Final diagnosis must depend upon a histological examination.

[Cytological aspects of various particular varieties of breast epitheliomas]. / Aspect cytologique de quelques variétés particulières d'épithéliomas mammaires

Needle aspiration for cytological investigation has been used at the Foundation Curie since 1954. Until 1973, no less than 3,176 mammary tumours have been studied by this method and comparison made with the histological findings. In the whole group some relatively rare carcinomas have been encountered -- mucoid, medullary, adenoid cystic, squamous and apocrine. Each category is described from the cytologic viewpoint and illustrated with an appropriate cyto/histologic example to show what a high degree of precision may be attained by diagnostic cytology in mammary cancer.

[Value of the cytologic diagnosis of adenopathies by aspiration biopsy. Cytohistologic comparison of 1756 cases]. / Valeur du diagnostic cytologique des adénopathies par ponction aspiration. Confrontation cyto-histologique de 1756 cas

The value of the cytologic diagnosis by aspiration needle biopsy is analysed in a series of 1756 adenopathies, verified by histological examination (1311 malignant nodes and 445 inflammatory or normal lymph nodes). The result is that a large number of lymph node tumours, either primary or secondary, are identified by the cytologic examination (from 77% to 91% of the cases according to the explored zone). Furthermore the virtually non existent percentage of false positive results (3% of the cases) proves the great accuracy of this method in the diagnosis of malignancy. In contrast, the diagnosis of benign nature must never be stated without biopsy (2% to 4% of the cases are erroneous by negative according to the explored zone). Indeed, even if benign cells are obtained, nothing allows to eliminate a malignant tumour or a limited inflammatory zone, not reached by the needle.

[Basocellular epithelioma of the back with metastases of the axillary ganglia]. / Epithélioma basocellulaire du dos avec métastase ganglionnaire axillaire

In a 52-year old woman, basal cell carcinoma of the back skin recurs after excision and the axillary lymph nodes are involved. This case is to be added to the list of basal cell carcinomas with metastatic evolution. The histologic study of a series of sections allows to show the migration of neoplastic cells in the vessels. Though exceptional, the ability of the basal cell carcinomas to yield metastases must be taken into account.

[Fine-needle aspiration biopsy and analysis of estrogen receptors in breast cancer]. / Ponction-aspiration à l'aiguille fine et dosage des récepteurs d'oestrogènes dans le cancer du sein.

Estrogen receptor concentration was measured in fine needle aspirates from 98 human mammary carcinomas. We performed a biochemical radiolabeled assay by the Charcoal-Dextran procedure on cytosols obtained after homogenization in a saline hypertonic buffer (0,4 M KCl) and centrifugation at 105,000 g. DNA content of the pellets was measured by fluorimetry. A sample of about 10(6) cells, containing at least 10 micrograms of DNA, was needed to obtain a reliable result. In this report, 74 samples were over 10 micrograms of DNA each. The analysis of these samples showed that, in 39 cases (52,8%), the estrogen receptor level was over the limit of 500 fmol/mg of DNA. In 5 cases (6,7%), the range was between 500 to 600 fmol/mg DNA and, in 30 cases (40,5%), it was under 500 fmol/mg DNA and was considered as negative. For 15 patients, estrogen receptors were determined on the same tumor, as removed by fine needle aspiration and by a surgical procedure (6 cases) or drill-biopsy (9 cases). There was a good correlation between the receptor content in needle aspirates and in tissue specimens for 13 cases out of 15.

[Metastatic breast cancer: a comparative study of the efficacy of tamoxifen and the sequential administration of tamoxifen and medroxyprogesterone acetate]. / Cancer du sein métastasé: étude comparée de l'efficacité du tamoxifène et d'une administration séquentielle de tamoxifène et d'acétate de médroxyprogestérone.

Seventy-nine patients with an histologically proven disseminated breast cancer, never treated before with additive hormonal therapy, entered into a randomized trial between june 1981 and december 1982. In the first group 44 patients were given continually a daily dose of tamoxifen (TAM) of 20 mg/m2. In the 2nd group 35 patients were given a daily dose of TAM of 20 mg/m2 for 15 days and then an oral daily dose of medroxyprogesterone acetate of 350 mg/m2 for the next 15 days. In both groups I and II, the treatment was stopped at the first manifestation of progression of the disease. The hormonal receptor status was determined in 30 patients of the group I and 23 patients of the group II. An objective response to treatment was observed in 48 per cent of the patients of the group I and 60 per cent of the group II. This difference is not significant (X2 = 1,05). However, the mean duration of therapeutic response is significantly higher in the group II (p = 0,01).

[Breast cancer. Development of the concentration of hormonal receptors under cytotoxic chemotherapy]. / Cancer du sein. Evolution de la concentration en récepteurs hormonaux sous chimiothérapie cytotoxique.

38 patients with locally advanced breast cancer, were treated with a program of cytotoxic chemotherapy including Adriamycin, Cyclophosphamide, 5 Fluoro-uracile and Prednisone. The concentration of hormonal receptors (Estrogen ER and Progesterone receptor PR) in the tumor were studied before starting the chemotherapy and after one to 9 courses of chemotherapy. This therapeutic response rate was not related to the initial level of ER and PR. After chemotherapy, we observed an increase of ER level in 14 cases out of 29 (59%), of PR level in 17 out of 32 cases (53%); A therapeutic response is observed in 11 cases out of 17 when the PR concentration is increased and in 3 cases out of 15 when the PR concentration was either stable or decreased. These observations let us to suppose a selective initial activity of chemotherapy on lesser differentiated tumor cells.

[Cytological examination. Solid breast tumors examined by means of fine-needle puncture]. / Examen cytologique. Tumeurs solides du sein par ponction à l'aiguille fine

The value of cytological diagnosis of solid tumours of the breast where the cells were obtained by suction aspiration through a needle has been analysed in a consecutive series of 2,401 cases, all of which were checked by histological examination. They consisted of 1,745 malignant tumours and 656 benign tumours. The cytological diagnosis of malignancy was confirmed, almost always, error being practically non-existent (4 out of 1,000). This confirms that this simple and rapid test is very valuable in the diagnosis of mammary cancer. On the other hand, when the diagnosis of a benign tumour was made a certain number of cancers were missed (3.6 per cent). This is not therefore as valuable a test when the result is negative as when the result is positive. All the same, in certain cases it can suggest the diagnosis without giving the precise type of lesion, which can only be finally diagnosed histologically.