Autocrine function for insulin-like growth factor I in human small cell lung cancer cell lines and fresh tumor cells.

We showed previously that insulin-like growth factor I (IGF-I) is detectable in small cell lung cancer (SCLC) tumor biopsies and cell lines and that recombinant human IGF-I stimulates DNA synthesis in SCLC cells. Here we report further studies on the role of IGF-I in 2 SCLC cell lines: HC12, classic; and ICR-SC17, variant. Immunoreactive IGF-I was detected in medium conditioned by HC12 but not ICR-SC17. Both HC12 and ICR-SC17 bound IGF-I with 100-fold greater affinity than insulin. Scatchard analysis revealed two classes of IGF-I binding site of high (Kd 0.1 nM, n = 2,300) and lower (Kd 3 nM, n = 28,000) affinity. In both cell lines [3H]thymidine incorporation was enhanced by recombinant human IGF-I, 100-1000 ng/ml. ICR-SC17 also showed growth enhancement as measured by increase in cell numbers. There was no response in HC12, probably due to endogenous IGF-I production. 125I-IGF-I binding and basal and IGF-I-stimulated mitogenesis were inhibited by monoclonal antibodies to IGF-I (SM1.20B, SM1.25) or the type I IGF receptor alpha IR3 but not an isotypic control monoclonal antibody. Antiproliferative effects were manifest in [3H]thymidine incorporation assays in serum-free conditions and growth of serum-supplemented liquid cultures. We also tested fresh or newly cultured tumor cells obtained by fine needle aspiration of metastases in three previously untreated and four relapsed patients with SCLC. IGF-I binding sites were demonstrable on fresh SCLC cells, and specific binding was inhibited by SM1.20B. All seven samples showed stimulation of [3H]thymidine incorporation in the presence of recombinant human IGF-I, 100-500 ng/ml. As in cultured cells, basal and IGF-I-stimulated DNA synthesis was inhibited by monoclonal antibodies SM1.20B, SM1.25, and alpha IR3 but not the isotypic control. These results confirm the findings of previous studies and suggest that IGF-I can function as an autocrine growth factor in SCLC in vitro and possibly also in vivo.

Presurgical determination of estrogen receptor status using immunocytochemically stained fine needle aspirate smears in patients with breast cancer.

We have developed an immunocytochemical staining procedure (ERICA) using a monoclonal antibody to the estrogen receptor (ER) to determine ER status from samples obtained by fine needle aspiration of primary and recurrent breast cancer tissue (cyto-ERICA). ER status was assessable on 214 of 246 smeared aspirates from breast cancer patients. In 143 (66.8%) assessable smears positive nuclear staining was observed but was completely absent in 71 (33.2%) cases. In 107 cases we were able to compare results with those obtained with the quantifiable dextrancoated charcoal (DCC) radioligand binding technique using surgically excised material. We observed qualitative agreement in 53 of 62 (85.5%) of primary specimens and 16 of 16 (100%) recurrent samples compared to the subsequent DCC result on the same sample. Aspirates obtained from new secondary deposits were also assessed and in 16 of 19 (84.2%) cases results agreed with that established previously by DCC on the primary breast tumor. In a further 6 of 10 (60%) cases the cyto-ERICA result obtained from recurrent samples qualitatively agreed with that determined by DCC on a previous recurrent lesion. A comparison of staining of aspirates was also made against frozen tissue sections stained with the monoclonal antibody (tissue-ERICA). Where comparison was made of primary tumor specimens agreement was observed in 40 of 45 (88.9%) of cases while specimens from secondary lesions agreed qualitatively in 14 of 17 (82.3%) of cases. In a small number of samples where tissue-ERICA was performed on an earlier lesion to that aspirated for cyto-ERICA an agreement of 4 of 5 (80%) was observed. This technique shows good sensitivity in demonstrating ER in aspirate specimens, should therefore permit us to determine ER status before surgery for primary breast cancer, and may also mean that surgery for recurrent disease to determine receptor status is no longer necessary.

Prediction of response to neoadjuvant chemoendocrine therapy in primary breast carcinomas.

Our aim was to determine whether biological molecular markers can predict response to neoadjuvant chemoendocrine therapy in patients with early breast cancer. Ninety patients (median age 56 years; range, 28-69 years) with primary operable breast carcinoma were studied. They were treated with four 3-weekly cycles of chemotherapy with mitozantrone, methotrexate (+/- mitomycin C), and tamoxifen prior to surgery. Fine-needle aspiration was used to obtain samples from patients prior to therapy, and the following parameters were assessed: estrogen receptor (ER), progesterone receptor (PgR), p53, Ki67, Bcl-2, and c-erbB-2 measured by immunocytochemistry, and ploidy and S-phase fraction (SPF) by flow cytometry. The tumors of 78% of the subjects responded (complete response, 9%; partial response, 69%) and 22% did not (no change, 20%; progressive disease, 2%). Response rates according to disease stage and patient age were as follows: T1, 74%; T2, 79%; T3/T4, 78%; age 50, 79% (P = not significant). Response rates for other parameters were as follows: ER-positive, 82%, and -negative, 70%; PgR-positive, 86%, and -negative, 71%; p53-positive, 74%, and -negative, 81%; Bcl-2-positive, 85%, and -negative 61%; c-erbB-2-positive, 57%, and -negative, 93%; Ki67 high, 77%, and low, 81%; SPF high, 77%, and low, 77%; aneuploid, 71%; and diploid, 85%. Only the difference for c-erbB-2 was statistically significant (P = 0.007). A trend for higher response rates to neoadjuvant chemoendocrine therapy for tumors that were positive for ER, PgR, and Bcl-2 was observed but did not reach statistical significance. Tumors negative for c-erbB-2 had a higher response rate, which was statistically significant. In contrast, Ki67, ploidy, SPF, and p53 failed to predict for response.

Use of soluble swabs in the diagnosis of bladder neoplasia.

A technique is described for obtaining samples of bladder epithelium using soluble swabs. Changes indicating neoplasia were seen in a large proportion of patients with bladder carcinoma. These changes were absent in the non-neoplastic control cases. The method provided tissue similar to that obtained by biopsy and surgical removal. The cellular appearances resembled those seen in neoplastic cells exfoliated into urine.

An ultrastructural study of the urinary bladder in children correlated with histological, bacteriological, and clinical findings.

Bladder biopsies from six boys without a history of urinary tract infection were taken during hypospadias repair operations and examined by electron microscopy. The ultrastructure of the epithelium, which was presumed to represent the normal, is described in detail and compared with the reported findings in other mammalian species. The appearances are generally similar although in our material the luminal membrane of the superficial cells was thicker than that surrounding the other epithelial cells but not asymmetrical. We observed membrane-coating granules which have not been reported before in bladder epithelium. The appearances of the normal bladder are compared to those seen in 24 children with urinary infection. In the presence of acute infection in large heterogeneous secondary lysosomes which are present in the intermediate and superficial cells of the normal bladder are reduced in number and the amount of rough endoplasmic reticulum is increased.

Fine needle aspiration cytology.

Fine needle aspiration cytology is an inexpensive, atraumatic technique for the diagnosis of disease sites. This paper describes the technique and illustrates how it may be applied to the management of tumours throughout the body. The limitations of the method, the dangers of false positive reports, and the inevitability of false negative diagnoses are emphasised. In a clinical context the method has much to offer by saving patients from inappropriate operations and investigations and allowing surgeons to plan quickly and more rationally. It is an economically valuable technique and deserves greater recognition.

Squamous cell carcinoma of the breast: clinico-pathologic implications and outcome.

AIMS: Pure squamous cell carcinoma (SCC) of the breast is a rare tumour and little is known about long-term outcome. We report our experience of a consecutive series of patients. METHODS: All patients with SCC treated at our institution between 1970 and 2001 were included. The pathological features, outcome and prognosis were studied. RESULTS: Eleven patients were identified. The median age was 55 (38-90) years and median follow-up was 62 (3-332) months. Four tumours were T1, three were T2 and three were T3 (one tumour size was unknown). There were seven poorly differentiated and three moderately differentiated SCC. Tumour grade could not be assessed in one patient. Primary treatment was mastectomy in six patients, wide local excision in four patients and radiotherapy in one patient. There was lymph node (LN) involvement in two patients. Oestrogen receptor status was assessed in seven patients and only one tumour was positive. Adjuvant chemotherapy was given to three patients and five patients received adjuvant radiotherapy. Two patients developed local recurrence at 5 and 12 months and three patients developed distant metastasis at 2, 36 and 306 months. Three patients were treated with chemotherapy at recurrence. Three patients have died of the disease, two are alive with disease and six remain well. The 2- and 5-year overall survival was 80% (SE=13%) and 67% (SE=16%) respectively. Large tumour size and positive LN status were prognostic indicators of poor outcome. CONCLUSION: SCC of the breast adopts an aggressive course with outcome comparable to poorly differentiated breast adenocarcinoma.

Gynaecological cytology and pelvic ultrasonography in patients with breast cancer taking tamoxifen compared with controls.

The objective of this study was to look at the effect of tamoxifen on the endometrium by comparing gynaecological cervical and endometrial cytology in breast cancer patients on tamoxifen for 3 years compared with controls. In addition, pelvic ultrasonography was employed to detect ovarian abnormalities and to measure endometrial thickness. Patients followed-up after primary surgical therapy for breast cancer were invited for gynaecological assessment consisting of clinical examination pelvic ultrasonography and a cervical smear and endometrial sampling. The patients taking tamoxifen (n = 49) has been on adjuvant hormone therapy for a minimum of 3 years. Control patients (n = 45) were also being followed-up for breast cancer. On examination the tamoxifen patients had very similar clinical findings to the control patients with regard to the cervix (normal in 84% of tamoxifen takers compared to 87% of controls). The uterus was clinically enlarged in eight patients on tamoxifen and in none of the control patients (P = 0.006) and only one ovarian cyst was clinically detectable in a patient taking tamoxifen. Pelvic ultrasonography between the two groups of patients was not statistically different (chi 2 test) and ovarian cysts were noted in nine patients from each group (tamoxifen patients 18% vs control patients 20%, n.s.). There was a highly significant difference in endometrial thickness in premenopausal patients (9.2 mm) compared with postmenopausal patients (6.4 mm, P = 0.001). There was also a suggestion that endometrial thickness was greater in tamoxifen treated patients (P = 0.08). In general, a greater proportion of patients taking tamoxifen had cervical and endometrial cells exhibiting hyperplastic nuclei, and in endocervical smears, this difference achieved statistical significance (Mann-Whitney test, P = 0.046). These findings show that a significantly increased proportion of patients taking tamoxifen had endocervical nuclear hyperplasia, and a trend towards increased endometrial thickness. These findings confirm that tamoxifen has mild oestrogenic activity. However, the lack of any difference in the incidence of dysplasia suggests that the carcinogenic potential of tamoxifen on the uterus is very low and the beneficial effects of tamoxifen as an adjuvant therapy for breast cancer outweighs its theoretical risks.

A retrospective study comparing the individual modalities of triple assessment in the pre-operative diagnosis of invasive lobular breast carcinoma.

AIMS: Early invasive lobular breast carcinoma (ILC) is associated with few symptoms and signs. The individual sensitivity of clinical examination, mammography, ultrasonography, cytology and core biopsy have each been reported to be of limited value. The aim of this study was to evaluate the accuracy of triple assessment in the pre-operative detection of patients identified to have ILC from their surgical pathology. METHODS: Pure ILC was defined as tumours containing at least 90% lobular features. The triple assessment of 273 patients diagnosed primarily at our institution were reviewed. RESULTS: 87.5% of women were symptomatic and 12.5% were screen detected. The mean patient age was 59 (range 30-81) years and the median tumour size was 26 (range 5-110) mm. The main mammographic abnormalities were a spiculated lesion (33.3%), an ill-defined mass (33.3%) or architectural distortion (23.5%). The sensitivities for detecting ILC of each modality were: clinical examination (76.6%), mammography (79.8%), ultrasound examination (93.9%), fine-needle aspiration cytology (FNAC) (60.5%) and core biopsy (90.8%). Combining the three modalities of clinical examination, imaging and cyto/pathology increased the pre-operative detection rate of ILC. CONCLUSION: Triple assessment is useful in the diagnosis of ILC. As the features of ILC may be subtle, a high index of suspicion is required to facilitate early diagnosis.

The value of cervical cytology in detecting recurrent squamous carcinoma of the cervix postradiotherapy.

Twenty-six patients with central recurrent squamous carcinoma of the cervix following radiotherapy were identified from hospital records, 22 of which had cervical smears available for review, and are described here. As a second part to this study the cervical cytology results for 1987 and 1988 were compared with the clinical findings and eventual outcome. From a total of 384 smears in 1987 and 1988 there were 11 correctly identifying recurrence. Two smears, however, were false positive and two false negative. The overall sensitivity was 85% with a specificity of 99.5%. In addition three microscopic, operable recurrences were identified by cytology. In experienced hands routine smear cytology postradiotherapy is a reliable and useful addition to surveillance.

The role of routine urine and parametrial aspirate cytology in staging carcinoma of the cervix.

The role of routine urine and parametrial aspirate cytology as a means of refining the staging of carcinoma of the cervix was assessed in a series of 45 consecutive patients undergoing examination under anaesthetic and cytoscopy. Only 2/45 (4%) of urine samples were positive and 3/37 (8%) of parametrial aspirates were abnormal. In none of these patients was additional information obtained over that already available from cytoscopy, digital examination and routine radiology. These procedures have no role in the routine clinical staging of cervical carcinoma.

Fine needle aspiration of the larynx: a preliminary report.

A technique for endoscopic fine needle aspiration cytology (FNAC) of the larynx is presented and its use in a preliminary study of twelve patients is discussed.

Diagnosing breast carcinoma in young women.

OBJECTIVE: To assess the individual and combined diagnostic accuracy of clinical examination, mammography, and fine needle aspiration biopsy in young women with breast cancer. DESIGN: Analysis based on case notes of patients presenting with breast cancer during 1971-89. SETTING: A combined breast clinic. PATIENTS: Consecutive series of 81 women aged less than 36 with histologically proved breast cancer presenting with a discrete mass over 19 years. MAIN OUTCOME MEASURES: Results of clinical examination, xeromammography or conventional mammography, fine needle aspiration biopsy, and examination of tissue removed by surgery. RESULTS: The clinical diagnosis was correct in 47 women and radiography in 35. Fine needle aspiration biopsy was correct in 47 of the 63 women in whom it was successfully performed. Fine needle aspiration was significantly more accurate than mammography (78% v 45%, p less than 0.01). Ten (16%) patients had negative results on clinical examination, mammography, and fine needle aspiration. CONCLUSION: Mammography alone seems inadequately sensitive to detect breast cancer in young patients. When all investigations give negative results excision biopsy is the only way of obtaining a definitive diagnosis.

Ureteric urine examination.

Uterteric urine examination can give definitive evidence of upper urinary tract carcinoma and three cases are described and illustrated. The absence of carcinoma cells in such specimens, however, does not exclude malignancy for two reasons. First, the tumours may be well differentiated and the cells not recognized at cytological examination and, secondly, carcinoma cells may not appear in ureteric urine specimens since they can be reluctant to exfoliate.

Needling renal cysts and tumours: cytology and radiology.

Renal masses found by intravenous urography, ultra-sound scanning, and arteriography were needled in 102 patients. Simple renal cysts containing clear fluid and no cytological abnormalities were found in 85 patients. Two unsuspected renal cell carcinomas were found on puncture; cytological examination showed malignant cells in the aspirate. Another five renal tumours were needled deliberately before nephrectomy, and a firm preoperative diagnosis of renal cell carcinoma was made on aspiration cytology in three. Benign cysts which had bled were particularly hard to diagnose. With care, radiology and cytology in combination can provide the firm diagnostic base needed for sound clinical management. The radiology-cytology team must be alert to the unusual finding that indicates a complex lesion, such as an unsuspected renal tumour.

Comparison of two methods of storing breast fine needle aspirates (FNAs) using oestrogen receptor immunocytochemical assay as a method of evaluating the storage methods.

Two methods of storing fine needle aspirates were compared in 14 patients with breast cancer. The methods of storage were: (1) as a Cytospin slide prepared immediately from the aspirated material and stored at -80 degrees C; (2) as a suspension of cells in tissue culture medium, stored at -80 degrees C. The effect of storage on the cells was assessed by means of an oestrogen receptor immunocytochemical assay (ER-ICA). An ER positivity of 100% was obtained by ER-ICA staining of cells after storage method 1, whilst all of the specimens stored by method 2 were ER-negative. The data demonstrate that cells stored in tissue culture medium at -80 degrees C are not suitable for ER measurement. The storage method of choice for specimens intended for ERICA is as a Cytospin slide. The ER status of cells deposited on Cytospin slides prepared immediately and stored at -80 degrees C for 2 years could be demonstrated despite the delay in processing the specimen.