Biomedical subjects

W Jonat

Publications and source records attributed to W Jonat.

At least 55 records · Page 3Linked to original sources

PubMed1999

Immunohistochemical assessment of p170 provides prognostic information in endometrial carcinoma.

AIMS: To determine the significance of proliferative activity (PA) in endometrial carcinomas, we analysed the expression of cell cycle-related antigens in routinely processed tissue. METHODS AND RESULTS: Serial sections of 164 endometrial carcinoma specimens were immunostained with the monoclonal antibodies (MoAb) Ki-S4 (specificity for p170) and Ki-S5 (specificity for p320/350). The Ki-S4 PA (median value 18.3%) and Ki-S5 PA (median value 25.0%) in endometrial carcinomas showed a significant correlation (r = 0.89, P < 0.001). A significantly lower median value of Ki-S4 and Ki-S5 immunoreactivity was ascertained in endometrial carcinomas stage I vs. > I (P < 0.05), in tumours with myometrial invasion < or = 50% vs. > 50% (P < 0.03), and in grade 1 vs. grade 2 and 3 tumours (P < 0.001). Univariate analysis showed that age, FIGO stage and grade, histopathological subtype, myometrial invasion, Ki-S4 and Ki-S5 PA provided prognostic information on the adjusted overall survival, whereas FIGO stage (P = 0.002) and Ki-S4 PA (P = 0.008) were independent prognosticators for adjusted overall survival in multivariate analysis. CONCLUSIONS: The association with established prognosticators for endometrial carcinomas, and the results of uni- and multivariate analysis indicates that the additional evaluation of Ki-S4 PA is useful for classifying patients into subgroups with low and high risk of relapse which might help individualizing the therapeutic strategy in endometrial cancer patients.

Adult↗

PubMed1998

Analysis of telomerase expression and proliferative activity in the different layers of cyclic endometrium.

To gain better insights into cell kinetics under physiological conditions, telomerase activity in the functional and basal layers of cyclic endometrium (n = 33) was compared with the immunostaining of glandular and stromal cells within these layers (n = 25). Two immunohistochemical proliferation markers were used to demarcate cells in the G1 phase of the cell cycle. In contrast to previous expectations, telomerase activity and both glandular and stromal proliferative activities were all significantly higher in the functional than in the basal endometrium (P < 0.002). The course of telomerase activity in the endometrial layers during the ovarian cycle was significantly associated with the proliferative scores for the functional and basal endometrial glands and the functional stroma but not the stromal compartment of the basal layer. Our findings indicate that the telomerase activity in cyclic endometrium is associated with the total number of proliferating glandular and stromal cells in the functional layer. Proliferating daughter cells of telomerase-competent stem cells may account for the lower levels of telomerase detected in normal basal endometrium.

Adult↗

PubMed1998

Anastrozole versus megestrol acetate in the treatment of postmenopausal women with advanced breast carcinoma: results of a survival update based on a combined analysis of data from two mature phase III trials. Arimidex Study Group.

BACKGROUND: This report presents the results of a survival update based on the combined data from two studies that compared the efficacy and tolerability of anastrozole (1 or 10 mg once daily), a selective, nonsteroidal aromatase inhibitor administered orally, and megestrol acetate (40 mg 4 times daily) in the treatment of postmenopausal women with advanced breast carcinoma whose disease had progressed after treatment with tamoxifen. METHODS: Two randomized, parallel-group, multicenter trials were conducted, involving a total of 764 patients. The two trials were identical in design; both were double blind for anastrozole and open label for megestrol acetate. Overview analyses were conducted with the intent of strengthening the interpretation of results from each trial. The median follow-up duration for this survival update was 31 months. RESULTS: At the clinical dose of 1 mg daily, anastrozole demonstrated a statistically significant survival advantage over megestrol acetate, with a hazard ratio of 0.78 (P < 0.025)(0.60 < 97.5% confidence interval [CI] <1.0). The 1 mg anastrozole group also had a longer median time to death (26.7 months) compared with 22.5 months for the megestrol acetate group. The 10 mg anastrozole group also had a survival benefit over the megestrol acetate group, with a hazard ratio of 0.83 (P=0.09, not significant)(0.64 < 97.5% CI < 1.1). Higher 2-year survival rates were observed for both anastrozole treatment groups than for the megestrol acetate group (56.1%, 54.6%, and 46.3% for the groups given 1 mg anastrozole, 10 mg anastrozole, and megestrol acetate, respectively). CONCLUSIONS: This combined analysis of two trials of postmenopausal patients with advanced breast carcinoma has clearly demonstrated that, after disease progression with tamoxifen, treatment with anastrozole 1 mg once daily results in a statistically and clinically significant advantage over a standard treatment, megestrol acetate. This important benefit, in addition to the good tolerability profile of anastrozole, supports the use of this drug as a valuable new treatment option for this patient population.

Aged↗

PubMed1998

Telomere shortening in uterine leiomyomas.

OBJECTIVE: To gain a better understanding of proliferation control mechanisms in a common benign tumor, we investigated the mean telomere length and the clonality of uterine leiomyomas. STUDY DESIGN: Deoxyribonucleic acid from uterine leiomyomas and from the adjacent normal myometrium of 51 patients (total number of uterine leiomyomas 107; 28 patients with single leiomyoma, 23 patients with multiple leiomyomas ranging from 2 to 8 myoma nodules per case) was hybridized to a telomeric oligonucleotide probe by Southern blot and chemiluminescent detection. The mean telomere length was evaluated by densitometry. Clonality was assessed with use of the phosphoglycerokinase gene polymorphism. RESULTS: The mean telomere length was significantly shorter in uterine leiomyomas (median 7950 bp, interquartile range 7261 to 8372 bp) than in normal myometrium (median 9688 bp, interquartile range 8528 to 10535 bp) (P < .001). There was no correlation between tumor size and telomere attrition. Multiple uterine leiomyomas were found to have an independent clonal origin. CONCLUSIONS: Telomere attrition in uterine leiomyomas reflects enhanced proliferation activity in the course of tumor evolution. The basic telomere lengths differ in the myocytes from which the uterine leiomyomas originate, probably explaining the lack of correlation between telomere attrition and tumor size.

Adult↗

PubMed1998

Influence of treatment with onapristone on the IGF-system in breast cancer patients.

The influence of the novel antiprogestin onapristone on the serum insulin-like growth factor (IGF) system was studied in a group of 13 postmenopausal women with metastatic breast cancer. Blood samples were obtained before treatment and subsequently after 1, 2 and 3 months on therapy. IGF-I, IGF-II and IGF-binding protein (IGFBP)-2 were measured by radioimmunoassay (RIA). In addition, the IGFBP profile was evaluated by Western ligand blotting (WLB), and IGFBP-3 fragmentation determined by immunoblotting. A moderate (29%) but significant increase in IGF-I was observed after 3 months on treatment (p < 0.05). IGFBP-2 showed a significant, progressive increase during treatment when evaluated both by WLB (44% increase over baseline at 3 months) and by RIA (33% increase over baseline at 3 months). There was a non-significant trend towards an initial decrease in IGFBP-3 fragmentation. No significant alterations were observed in IGF-II or any of the binding proteins (except IGFBP-2) determined by Western ligand blotting. Due to the observation that onapristone treatment caused a moderate suppression of serum cortisol and androstenedione, we postulate the observed increase in IGF-I to be due to a slight glucocorticoid agonistic effect of the drug. On the contrary, the increase in IGFBP-2 may be related to disease progression as has been observed in patients suffering from prostatic cancer.

Adult↗

PubMed1998

Luteinizing hormone-releasing hormone analogues--the rationale for adjuvant use in premenopausal women with early breast cancer.

Current standard adjuvant therapies for early breast cancer include tamoxifen and chemotherapy, depending on the disease prognosis and menopausal status. Luteinizing hormone-releasing hormone (LHRH) analogues offer a different approach to the management of early breast cancer in pre- and perimenopausal women. The most widely studied LHRH analogue is goserelin. It acts on the hypothalamic-pituitary axis to suppress ovarian function, decreasing luteinizing hormone and oestradiol levels to post-menopausal values. Pooled data from 228 premenopausal and perimenopausal patients with advanced breast cancer enrolled in 29 studies worldwide demonstrated an objective response rate for goserelin, 3.6 mg, of 36.4%, with a median duration of response of 44 weeks. These results fall well within the ranges of reported response rates for ovarian ablation and for tamoxifen in similar patient populations. By virtue of its mode of action, goserelin does not stimulate the ovaries and is unlikely to have detrimental effects on the endometrium. In addition, given that goserelin has no oestrogen agonist-like effects, unlike tamoxifen, there is no potential for tumour stimulation in those patients becoming resistant to treatment. Goserelin is generally well tolerated, and the main side-effects are related to ovarian suppression, which is potentially reversible. Preliminary results in premenopausal women with early breast cancer indicate that endocrine treatment with goserelin plus tamoxifen may be as effective as standard combination chemotherapy (cyclophosphamide-methotrexate-5-fluorouracil), but has significantly less acute toxicity. A number of large, randomized trials are now in progress to assess the potential role of goserelin as adjuvant therapy for early breast cancer.

Antineoplastic Agents, Hormonal↗

PubMed1998

One year of experience working with the aid of a robotic assistant (the voice-controlled optic holder AESOP) in gynaecological endoscopic surgery.

The aim of the study was a comparison of robotic versus human laparoscopic camera control. Utilizing robotic technology a robot has been designed specifically for the purpose of holding and manoeuvring the laparoscope under the direct control of the surgeon. We tested AESOP (automated endoscopic system for optimal positioning) in 50 patients undergoing routine gynaecological endoscopic surgical procedures. The elimination of the camera holder allows two doctors to perform complex laparoscopic surgery faster than without the robotic arm. The timing of surgical procedures performed by surgeons using the voice control was compared to the timing of similar operations using the foot or hand control. The voice-controlled AESOP works more efficiently and faster than the hand or foot control.

Endoscopes↗

PubMed1998

[Intact pregnancy with concomitant hydatidiform mole of a coexisting placenta].

The simultaneous occurrence of an intact pregnancy with normal placenta morphology and sonographic signs of a hydatidiform mole of a second placenta, which was diagnosed in the 14th gestational week, is described. Prenatal diagnostic and perinatologic management will be reported. The difference of the placentas will be shown by histopathology.

Adult↗

PubMed1998

Sonographically detected fetal and placental abnormalities associated with trisomy 16 confined to the placenta. A case report and review of the literature.

Trisomy 16 is the most frequent autosomal anomaly seen in early spontaneous abortions, accounting for 15 per cent of all chromosomally abnormal early spontaneous abortions. This trisomy is thought to be lethal in the non-mosaic state and incompatible with full fetal development. We report a case of placental trisomy 16 mosaicism detected after chorionic villus sampling (CVS). The patient was referred at 18 weeks of gestation on account of moderate intra-uterine growth restriction (IUGR). Detailed sonography showed a thickened and enlarged placenta with multiple 'cysts', polyhydramnios, a single umbilical artery and a small ventricular septal defect (VSD). CVS, amniocentesis (AC) and fetal blood sampling (FBS) were performed. After direct preparation of chorionic villi only 47,XX,+16 cells were seen. However, chromosomal analysis of cultivated amniotic fluid cells and fetal lymphocytes only showed a normal karyotype 46,XX. After direct preparation of a second CVS at 19 + 4 weeks of gestation the karyotype 47,XX+16 was confirmed in the contralateral part of the placenta and near the insertion of the umbilical cord. A normal female karyotype 46,XX was demonstrated by extensive karyotyping of various sites of the placenta, the neonatal skin fibroblasts and lymphocytes postnatally. In accordance with this observation the multiple 'cysts' of the placenta disappeared in the third trimester. We speculate that the sonographic findings of multiple round placental 'cysts' without a hyperreflective border may be caused by the trisomic cell lineages and therefore may be a sonographic marker of trisomy 16.

Abnormalities, Multiple↗

PubMed1997

p53 is a persistent and predictive marker in advanced ovarian carcinomas: multivariate analysis including comparison with Ki67 immunoreactivity.

p53 mutation and p53 protein overexpression are common findings in ovarian carcinomas. In order to evaluate the prognostic significance of the p53 status and its role in metastasis, we examined 104 ovarian carcinomas, among them 83 cases with follow-up data, and 40 pairs of primary tumors and metastases, by p53 immunohistochemistry and temperature-gradient gel electrophoresis. Comparison of primary tumors and their metastases revealed identical results in 88%-90% of the cases, indicating that, in most cases, mutant p53 occurs prior to metastatic spread and remains clonally conserved. With respect to all tumors, moderate/high p53 expression was significantly more prevalent in serous-papillary types, carcinomas with high grade, and high Ki67 scores, but was not associated with age, stage, or hormone receptor status. Kaplan-Meier analysis of 83 cases, followed-up for 9-96 months, demonstrated that moderate/high p53 overexpression in the group of 66 stage T3/M1 tumors was associated significantly (P = 0.0028 and P = 0.0105) with shorter overall and recurrence-free survival. Multivariate analysis revealed that advanced clinical stage and p53 positivity were the only independent predictive variables. No significance was seen in regard to second-look results and outcome of 50 patients receiving platinum-based chemotherapy. These observations show that p52 immunohistochemistry is an independent prognostic indicator at the given cut-off level, but does not reliably predict chemotherapy response.

Adult↗

PubMed1997

Clinical overview of anastrozole--a new selective oral aromatase inhibitor.

The efficacy and tolerability of the new selective aromatase inhibitor, anastrozole (Arimidex), was compared with megestrol acetate in the treatment of advanced breast cancer in postmenopausal women. In two independent prospective randomised trials, patients who progressed after prior tamoxifen therapy received anastrozole 1 or 10 mg once daily, or megestrol acetate, 40 mg q.i.d. The two studies were designed to allow the data to be combined to increase the statistical power of the analyses. It is the data from these combined analyses that are considered in further detail. After 6 months of follow-up, the proportion of patients gaining clinical benefit (complete response + partial response + stable disease > or = 24 weeks) was approximately one third of patients in all three groups. No significant difference was observed between either dose of anastrozole and megestrol acetate in the time to disease progression (130-153 days). All three treatments were generally well tolerated, but significantly more patients on megestrol acetate gained weight, and the weight gain in this group continued up to at least 9 months of follow-up. At a 12-month update of tolerability, of the commonly observed adverse events, a greater than 2-fold difference between treatment arms was observed for hypertension, weight gain, dyspnoea, vaginal haemorrhage, sweating and diarrhoea (all higher on megestrol acetate except for diarrhoea). Anastrozole is effective and well tolerated and on the basis of these data, 1 mg once daily is the recommended clinical dose in postmenopausal women with advanced breast cancer.

Administration, Oral↗

PubMed1997

Fetal heart rate (FHR) pathology in labor related to preceeding Doppler sonographic results of the umbilical artery and fetal aorta in appropriate and small for gestational age babies. A longitudinal analysis.

The aim of this study was to ascertain the value of serially performed Doppler sonographic measurements of fetal vessels for the prediction of FHR alterations in labor a longitudinal analysis was conducted. 24 patients with SGA fetuses as the only risk factor and 38 patients without any risk factor were recruited for the study. Flow velocity waveforms of the fetal aorta and the umbilical artery were analyzed for systolic diastolic (S/D) ratio weekly at 20-39 weeks gestation and from 30 weeks gestation onwards twice weekly. Courses were related to complications during labor reflected by alterations of FHR tracings. The more numerous pathologie S/D ratios of both fetal vessels were recorded the more frequently complications in labor occurred (Chi Square test, p < 0.05). The mean value of the S/D ratios in fetuses with FHR pathology in labor differed significantly compared to the uncomplicated group (Wilcoxon rank sum test, p < 0.05). The fluctuation of S/D ratios was greater in the complicated than in the normal group (Wilcoxon rank sum test, p < 0.05). A combination of parameters describing S/D ratios showed a sensitivity of 86% and a specificity of 19% for the fetal aorta and a sensitivity of 71% and a specificity of 91% for the umbilical artery. Serial Doppler measurements of the fetal aorta and the umbilical artery aid in predicting pathologic FHR alterations in labor and may be of benefit in antenatal care to ensure fetal well being particularly in cases of IUGR.

Aorta↗

PubMed1997

ARIMIDEX: a potent and selective aromatase inhibitor for the treatment of advanced breast cancer.

Aromatase inhibitors have been available for a number of years and their ability to reduce circulating estradiol levels has been shown to produce clinical benefit in women with advanced breast cancer. Until recently, the only commercially available aromatase inhibitor was aminoglutethimide. Although aminoglutethimide has been shown to be efficacious in the treatment of advanced breast cancer, it does cause significant toxicity and requires the use of concomitant hydrocortisone therapy. Anastrozole is one of a new class of potent aromatase inhibitors able to suppress estradiol to the limit of detection of sensitive assays without suppressing adrenal steroidal synthesis. Two large clinical trials (n = 764) conducted in the U.S.A. and in Europe evaluated two doses of anastrozole, 1 and 10 mg a day, compared to megesterol acetate, 40 mg four times a day, in postmenopausal women who had progressed while on tamoxifen. Response rates and time to progression with anastrozole were similar to those of megesterol acetate. Objective responses (CR + PR) were 10.3%, 8.9% and 7.9% in the 1 and 10 mg of anastrozole and megesterol acetate treatment groups, respectively. Another 25.2%, 22.6% and 26.1% had stable disease for over 24 weeks on 1, 10 mg anastrozole and megesterol acetate, respectively. Anastrozole and megesterol acetate were well tolerated; however, more patients had significant weight gain on megesterol acetate than with anastrozole treatment. The weight gain seen with megesterol acetate continued to increase over time. Anastrozole has a better therapeutic index (fewer side-effects) and has recently been approved by the FDA and a number of other regulatory agencies around the world for the treatment of advanced breast cancer.

Anastrozole↗

PubMed1996

Regulation and role of TGF beta production in breast cancer.

The influence of antiestrogens on the secretion of transforming growth factor beta (TGF beta) proteins that have an autoinhibitory potential for human cancer cells was studied in the estrogen-responsive human breast cancer cell line, MCF-7: Antiestrogens induce the secretion of TGF beta-1 via a nontranscriptional pathway; TGF beta-1 itself induces TGF beta-2 by a direct transcriptional mechanism; and TGF beta-2 is a marker of antiestrogen action. This hypothesis was confirmed in a clinical study with 18 patients with advanced metastatic breast cancer. TGF beta-2 plasma levels were measured before and after 4 weeks of treatment with tamoxifen. In the majority of patients who responded to the treatment, increasing TGF beta-2 concentrations were seen under therapy. Patients who did not respond did not show changes in the TGF beta-2 plasma level after 4 weeks of treatment. These results suggest that the sequential analysis of TGF beta-1 in plasma before and under treatment with tamoxifen allows the early identification of patients with antiestrogen resistance.

Breast Neoplasms↗

PubMed1996

Reverse transcriptase/polymerase chain reaction detection of cytokeratin-19 mRNA in bone marrow and blood of breast cancer patients.

A two-step reverse-transcriptase-based polymerase chain reaction (PCR) with nested primer pairs was developed to amplify sensitive and specific cytokeratin-19 (CK-19) mRNA sequences from human breast cancer cells. No CK-19 pseudogene interference was seen. The larger DNA-derived amplification products could be clearly discriminated from mRNA-derived products. The CK-19 message was not amplified from bone marrow or blood of healthy volunteers and patients with haematological malignancies nor from myeloid and lymphoid cell lines. Breast cancer cells were diluted in buffy coat cells up to 10(-6) and CK-19 mRNA sought by PCR. The CK-19 message was detected in 14 of 26 blood samples and 14 of 24 marrow samples but in neither of two peripheral blood stem cell samples taken from 35 breast cancer patients. By sequence-analysis control of two of these samples and two cell lines, the amplified DNA fragments were confirmed to be homologous with the CK-19 sequence. The CK-19 message was further sought in matched blood/marrow samples taken from 13 untreated women in the same cohort at the time of diagnosis. In 3 of these, CK-19 RNA was detected in blood and marrow and, in 3 others, only in blood, but never in marrow alone. The results show that CK-19 assay by reverse transcriptase/PCR is a sensitive and specific technique for the detection of cancer cells in bone marrow and blood. It could be helpful in diagnosis and monitoring of metastatic breast cancer and detection of micrometastases. This should be evaluated on larger numbers of patients, with different clinical samples and epithelial malignancies.

Base Sequence↗

PubMed1996

A randomised trial comparing two doses of the new selective aromatase inhibitor anastrozole (Arimidex) with megestrol acetate in postmenopausal patients with advanced breast cancer.

The aim of this study was to compare the efficacy and tolerability of the new aromatase inhibitor 'ARIMIDEX' (anastrozole) with megestrol acetate in the treatment of advanced breast cancer in postmenopausal women. Anastrozole is a new potent and highly selective non-steroidal aromatase inhibitor. We conducted a prospective randomised trial comparing two doses of anastrozole (1 and 10 mg orally once daily) with megestrol acetate (40 mg orally four times daily) in postmenopausal patients with advanced breast cancer who progressed after prior tamoxifen therapy. All patients were analysed for efficacy as randomised (intention to treat) and for tolerability as per treatment received. Of the 378 patients who entered the study, 135 were randomised to anastrozole 1 mg, 118 to anastrozole 10 mg, and 125 patients to megestrol acetate. After a median follow-up of 192 days, response rate which included complete response, partial response and patients who had disease stabilisation for 6 months or more was 34% for anastrozole 1 mg, 33.9% for anastrozole 10 mg and 32.8% for megestrol acetate. There were no statistically significant differences between either dose of anastrozole and megestrol acetate in terms of objective response rate, time to objective progression of disease or time to treatment failure. The three treatments were generally well tolerated, but more patients on megestrol acetate reported weight gain, oedema and dyspnoea as adverse events while more patients on anastrozole reported gastro-intestinal disorders, usually in the form of mild transient nausea. Patients on anastrozole did not report higher incidences of oestrogen withdrawal symptoms. Anastrozole is an effective and well tolerated treatment for postmenopausal patients with advanced breast cancer. The higher 10 mg dose did not result in additional clinical benefit, but was well tolerated reflecting the good therapeutic margin with anastrozole. Based on this data, anastrozole 1 mg should be the recommended therapeutic dose.

Adult↗

PubMed1996

Anastrozole, a potent and selective aromatase inhibitor, versus megestrol acetate in postmenopausal women with advanced breast cancer: results of overview analysis of two phase III trials. Arimidex Study Group.

PURPOSE: To compare the efficacy and tolerability of anastrozole (1 and 10 mg once daily), a selective, oral, nonsteroidal aromatase inhibitor, and megestrol acetate (40 mg four times daily), in postmenopausal women who progressed following tamoxifen treatment. PATIENTS AND METHODS: Two randomized, double-blind for anastrozole, open-label for megestrol acetate, parallel-group, multicenter trials were conducted in 764 patients. Because both trials were identical in design, an analysis of the combined results was performed to strengthen interpretation of results from each trial. RESULTS: The median follow-up duration was approximately 6 months. The estimated progression hazards ratios were 0.97 (97.5% confidence interval [CI], 0.75 to 1.24) for anastrozole 1 mg versus megestrol acetate and 0.92 (97.5% CI, 0.71 to 1.19) for anastrozole 10 mg versus megestrol acetate. The overall median time to progression was approximately 21 weeks. Approximately one third of patients in each group benefited from treatment. Twenty-seven patients (10.3%) in the anastrozole 1-mg group, 22 (8.9%) in the anastrozole 10-mg group, and 20 (7.9%) in the megestrol acetate group had a complete or partial response, and 66 (25.1%), 56 (22.6%), and 66 (26.1%) patients, respectively, had stable disease for > or = 24 weeks. For all end points, individual trial results were similar to the results of the combined analysis. Anastrozole and megestrol acetate were well tolerated. Gastrointestinal disturbance was more common among patients in the anastrozole groups than the megestrol acetate group; the difference between the anastrozole 10 mg and megestrol acetate groups was significant (P = .005). Significantly fewer patients in the anastrozole 1-mg (P < .0001) and 10-mg (P < .002) groups had weight gain than in the megestrol acetate group. More than 30% of megestrol acetate-treated patients had weight gain > or = 5%, and 10% of patients had weight gain > or = 10%. Patients who received megestrol acetate continued to gain weight over time. CONCLUSION: Anastrozole, 1 and 10 mg once daily, is well tolerated and as effective as megestrol acetate in the treatment of postmenopausal women with advanced breast cancer who progressed following tamoxifen treatment. Moreover, anastrozole therapy avoids the weight gain associated with megestrol acetate treatment.

Adult↗

PubMed1996

Detection of cancer cells in peripheral blood stem cells of women with breast cancer by RT-PCR and cell culture.

The benefit of high-dose therapy and blood stem cell reinfusion for women with high-risk breast cancer is currently under investigation. Contaminations of autologous blood stem cells with cancer cells have been described. Cancer micrometastases may be detected by immunocytochemistry, culture techniques and cytokeratin-19 mRNA reverse transcriptase PCR. Women with breast cancer received adjuvant HD-CTM with peripheral blood stem cell (PBSC) support after surgical therapy and 4 cycles conventional chemotherapy. Peripheral blood stem cells were mobilised by G-CsF and harvested after the third or fourth cycle of standard therapy. Aliquots of PBSC-collections (10(7)-2*10(7) cells) were subjected to CK19-mRNA reverse transcriptase PCR. RNA was extracted by standard methods and reverse transcription was performed with MMV-RT. Integrity of RNA was checked by coamplification of housekeeping sequences. Aliquots of the RT-mix were subjected to PCR-amplification with outer and inner primer pairs, subsequently. A second aliquot of 2*10(7) cells was cultured over 42 days in liquid culture. Cytospins were prepared weekly from cultured cells and evaluated by light microscopy with or without prior immunocytochemistry. Ten leukaphereses from 6 women were available for PCR-analysis and cell culture. Six leukaphereses were negative for CK19-mRNA and for detection of cancer cells by culture technique, two samples were positive for CK19-mRNA and culturally enriched cells and two samples were positive for CK19-mRNA and negative for cultured cancer cells. No sample was positive for cultured cells and negative for CK19-mRNA. Overall, the results corresponded in 80%. Two sensitive techniques for the detection of cancer micrometastases were applied to aliquots from 10 leukaphereses of six breast cancer patients with corresponding results in 80%. PCR-mediated detection of cancer cells was confirmed by culture technique and light microscopy, however, further comparison of CK19-PCR with standard techniques like cell culture and immunocytochemistry is still necessary.

Breast Neoplasms↗