H. Rauschecker

Therapy of small breast cancer: A prospective study on 1036 patients with special emphasis on prognostic factors

In 1983, The German Breast Cancer Study Group, sponsored by the Federal Ministry of Research and Technology, started a prospective multicenter trial on the treatment of early breast cancer pT1 pN0 M0. Treatment consisted of initial tumorectomy with microscopically free margins and lower axillary dissection. After conformation of a pT1 pN0-stage, additional treatment was either mastectomy or adjuvant radiotherapy (50 Gy in 25 fractions to the entire breast plus 12 Gy electron boost). In medially located tumors, the parasternal and supraclavicular area was also irradiated with 50 Gy. A randomization between both treatment modalities was initially planned but was not feasible and abandoned. Nearly all patients were treated according to their own choice. From November 1983 through December 1989, 1119 patients were recruited. Eighty-three were excluded from the protocol. Out of the remaining 1036 patients, 733 (71%) underwent breast preservation and 303 (29%) mastectomy. A detailed pathohistological examination of all tumorectomy specimens was performed in a pathologic reference center. Oncogen overexpression was evaluated by immunohistological detection of the transmembrane protein p-185 (corresponding to c-erb-B2) in 425 cases. After a median follow-up of 48 months, the frequency of local recurrences (4.7%), regional recurrences (1%), and distant metastases (5.4%) was the same in the breast preservation group and the mastectomy group. The 3-year disease-free survival was 90% after breast preservation and 88% after mastectomy (p = 0.21). In the breast preserving group, 24 patients with microscopically involved margins had a poorer disease-free survival than the study group (75% vs 90% after 3 years). The width of the margins had no impact on prognosis. Other prognostic factors in an univariate and multivariate analysis were tumor size and tumor grade. Age, menopausal status, hormone receptor status, histological tumor type, and treatment (mastectomy vs breast preservation) were not significant. P-185-expression was dependent on tumor grade and was the strongest prognostic factor in an univariate and multivariate analysis (p less than 0.001). The results emphasize the central role of tumor grade for prognosis and suggest the independent prognostic significance of the c-erb-B2 oncogen (corresponding to p-185) in pN0-patients.

Reverse transcriptase/polymerase chain reaction analysis of parathyroid hormone-related protein for the detection of tumor cell dissemination in the peripheral blood and bone marrow of patients with breast cancer

Tumor cell dissemination in the bone marrow is an independent prognostic marker for relapse and survival for patients with primary breast cancer. Parathyroid-hormone-related protein (PTHrP) is expressed in most primary tumors and bone metastases of patients with breast cancer. PTHrP acts as an autocrine growth factor for breast cancer cells in vitro and there is evidence that it is especially important for osseous metastasis. For a sensitive detection of PTHrP-positive disseminated tumor cells a reverse transcriptase/polymerase chain reaction (RT/PCR) assay for PTHrP transcripts in the peripheral blood (PB) and in the bone marrow (BM) has been established. In mixing studies, the sensitivity of the reverse transcriptase/polymerase chain reaction (RT/PCR) for PTHrP was one tumor cell in 1×106 mononuclear cells. At this level of sensitivity, transcripts of PTHrP were detected in none of 30 PB samples and in 3 of 25 BM samples of healthy volunteers: there were also no transcripts of PTHrP in the PB and BM of 6 patients with benign breast lesions. The PB samples of 31 patients and the BM samples of 34 patients with predominantly early-stage breast cancer were tested for PTHrP expression along with immunocytology against cytokeratin 18 (CK18) as a standard immunological detection technique. PTHrP expression was shown in 9 of 31 patients in the PB and in 9 of 34 patients in the BM. In 30 patients, PB and BM samples were available simultaneously. There were cases of combined positive findings in the PB and the BM (4/30) and of isolated positivity in the PB (5/30) or in the BM (4/30). Compared to immunocytology, RT/PCR assay of PTHrP assay was significantly more sensitive in the peripheral blood (8/30 by RT/PCR compared to 1/30 by immunocytology). In the bone marrow there were cases of positivity for both markers (2/34), cases of isolated positivity by immunocytology for CK18 (3/34) and cases of isolated positivity for PTHrP transcripts (7/34). In conclusion the RT/PCR assay for PTHrP transcripts is a feasible and very sensitive technique for the detection of tumor cell dissemination in the PB, even in patients with early-stage breast cancer. The specificity of detection of PTHrP transcripts in the bone marrow is limited, possibly because of autochthonous expression of PTHrP in osteoblastic cells. The clinical follow-up of the subgroups of patients at risk, as defined by this assay, will show its prognostic significance for patients with breast cancer.

Therapy of small breast cancer - four-year results of a prospective non-randomized study

AbstractBackground: In the early 1980s breast preservation was a rarely applied therapeutic modality in the primary treatment of breast cancer in the Federal Republic of Germany. Reports coming from retrospective studies as well as preliminary results from randomized trials made it desirable to introduce breast preservation in the form of a controlled clinical trial. Study design: In stage pT1 N0 M0 breast cancer, mastectomy as the standard treatment was to be compared with radiotherapy of the remaining breast tissue. The study design originally planned as a randomized trial had to be changed into a prospective observation study due to the low randomization rate. Univariate analysis of prognostic variables was the first step to a valid treatment comparison. Those factors determined as being significant were included together with the treatment effects in a multivariate analysis. A high therapeutic standard was guaranteed by quality control. Results: 1036 out of 1119 recruited patients are evaluable. After a median follow-up of 48 months the following preliminary results can be reported. With the exception of death without recurrence from breast cancer, the 143 events are evenly distributed among the two treatment groups. Locoregional recurrence of the whole patient population was 5%. Out of all prognostic factors examined only tumor size and grading are significant in regard to recurrent disease. Recurrence-free survival decreased in cases with ‘uncertain’ tumor margins, whereas the width of the margin had no influence on recurrent disease. There was no significant difference in quality of life between the two treatment groups. Conclusions: The four-year results of this study are in accordance with those of other breast preservation trials: There is no significant difference between the two treatment groups in the occurrence of locoregional failure. Incomplete tumorectomy has a negative influence on recurrence. Quality of life seems more dependent on the acceptance of the therapy by the patient than on the therapeutic modality itself. Breast preservation can also be performed appropriately in smaller institutions if the therapeutic standard is guaranteed by quality control.

A sensitive high-performance liquid chromatographic assay for melphalan and its hydrolysis products in blood and plasma

SummaryA sensitive high-performance liquid chromatographic assay has been developed for the measurement of the alkylating cytostatic drug melphalan (4-[bis(2-chloroethyl) amino]-l-phenyl-alanine, or l-phenylalanine-mustard, L-PAM) and its two hydrolysis products, monohydroxy melphalan (MOH) and dihydroxy melphalan (DOH). A reversed-phase phenyl column and a mobile phase consisting of acetonitrile/citrate buffer made possible an isocratic separation and quantification. N,N-[bis (2-hydroxy-ethyl)]toluidine has been synthesized as an internal standard structurally related to DOH. A new, accurate “kinetic” calibration procedure enabled us to determine even the concentration of the unstable MOH. The lower limit of quantification was 30 mg/ml for L-PAM and 20 ng/ml for both DOH and MOH with fluorescence detection. The use of this method is illustrated by some pharmacokinetic data in systemic and locoregional melphalan therapy.

Mastectomy stimulates prolactin release in breast cancer patients

Hyperprolactinaemia has been described to occur after mastectomy in breast cancer patients, but whether it may be the result of surgery or breast cancer is particularly unknown. Plasma prolactin levels were measured in 51 patients one day before, and 1, 7, 30, and 180 days after mastectomy (23 primary breast cancer patients), tumourectomy (10 patients with benign or malignant breast lesions), and cholecystectomy (18 patients with cholelithiasis). Elevated prolactin levels were found on the 7th and 30th postoperative day in mastectomized and laparotomized patients, but not in patients who underwent tumourectomy of benign or malignant breast lesions (p less than 0.01). The prolactin levels were in the normal range one day before and again 180 days after surgery in all patients. We conclude, therefore, that postoperative hyperprolactinaemia in breast cancer patients is a result of surgery rather than the disease.

Leakage measurement during selective limb perfusion using a gamma probe

The objective of this study was to establish a probe system for intraoperative quantitative leakage measurement during selective limb perfusion for adjuvant high-dose chemotherapy in patients with malignant melanomas. We used a portable gamma probe with digital display and investigated the physical properties in a phantom study simulating blood pool activity at different angles of the probe to the surface and different distances. In 20 patients the limb circulation was surgically separated from the systemic blood circulation, and the limb was then selectively perfused (cytostatics added) for 60 min. Initially, 15 MBq technetium-99m labelled autologous red blood cells was injected into the limb circulation, and an equal amount was kept as a standard. Every 10 min, blood samples were drawn from the body circulation and count rates were simultaneously measured by the probe system at the lower end of the sternal body. At the end of perfusion, the circulation of the limb was reconnected, the standard injected into the systemic circulation, and a blood sample drawn after 10 min. All blood samples were counted for calculation of leakage in terms of percent of the injected dose, and the results compared with the intraoperative count rates of the probe system. In the range of leakage observed in this study (0%–86%), the count rate of the probe system (corrected for blood volume, i.e. for body surface) correlated with the results of conventional measurement (r=0.92) according to the equation: %leakage=counts per sx[1.2×body surface (m2)−1.19]. In conclusion, the use of the described probe system is a feasible approach for leakage quantification which continuously yields data during selective limb perfusion.

Kinetics of melphalan leakage during hyperthermic isolation perfusion in melanoma of the limb

SummaryThe kinetics of melphalan leakage into the peripheral blood were studied in 21 patients undergoing hyperthermic isolation perfusion of the upper or lower limb as an adjuvant treatment in high-risk melanoma; in 5 patients cisplatin was added. The melphalan concentrations in the peripheral blood rose predominantly during the first 20 min of perfusion and levelled out to an apparent steady state of about 0.28 μg/ml in upper extremity perfusions, and 0.34 (without cisplatin) and 0.37 μg/ml (with cisplatin) in lower extremity perfusions. Erythrocytes labelled with technetium Tc 99m, which were added concomitantly with melphalan to the perfusion medium, appeared in the systemic circulation of the patients at an almost constant rate of 0.32% (lower and upper limb perfusions without cisplatin and 0.37% (with cisplatin) of total tracer/min. This perfusate flow rate indicated by labelled erythrocytes completely explained the leakage of melphalan from the perfusion circuit into the peripheral blood. Peak concentrations of melphalan in the peripheral blood were observed immediately after reconstitution of normal hemodynamic conditions once isolation perfusion had been teminated. This fraction of melphalan might originate from tissue-binding sites, but also from vascular compartments; therefore, a thorough washing-out procedure might minimize this effect.

Erniedrigte Plasmazinkspiegel beim metastasierten Mammakarzinom

Zinc is an essential component of many metalloenzymes for DNA and proteinsynthesis including RNA and DNA polymerases. It has been shown by several investigators that zinc is accumulated in breast cancer tissues. To investigate a possible relation between plasma zinc levels and tumor load, plasma zinc levels were evaluated in 76 patients with non metastatic breast cancer (no evidence for disease after mastectomy) and in 66 patients with metastatic breast cancer. Zinc concentrations were measured in plasma using an atomic absorption spectrophotometer (normal range 80-150 mcg/dl). In patients with metastatic disease plasma zinc concentrations were in the lower region of the normal range or depressed (arithmetic mean: 84.9 SD 21.6 mcg/dl), whereas patients with non metastatic breast cancer had normal zinc levels (arithmetic mean: 126.0 SD 27.7 mcg/dl). The difference between the two groups was highly significant (p = 0.001, t = -9.742, 140 degrees of freedom). It is concluded, that plasma zinc in breast cancer patients is depressed according to the stage of the disease. Based on experimental data a substitution of zinc cannot be recommended.

Determination of TPA levels in breast cancer and controls.

SummaryThe clinical significance of radioimmunological determination of Tissue Polypeptide Antigen (TPA) has been studied on patients with breast cancer (n=376), on “normal subjects” (n=92), on benign diseases of the breast as well as on patients with inflammatory diseases (n=98). TPA levels were elevated (120 U/l or higher) in the group with inflammatory diseases in 68% and in the group with breast cancer (stage IV with progression of disease) in 85%. In all other groups (healthy controls, benign diseases of the breast, breast cancer before operation, breast cancer stage I (NED), breast cancer stage II and III (NED), and breast cancer stage IV (PR/CR), TPA was higher only in 5–22%. TPA determinations seem not to be very useful for diagnostic purposes in breast cancer, but it can be regarded as suitable for monitoring proliferative processes in advanced breast cancer. Limitations result from lack of tumor specificty of the proliferation marker TPA. So far, follow-up studies after mastectomy in breast cancer and in patients with advanced breast cancer under chemotherapy have shown that CEA and TPA are concordant. There is no cross reactivity between CEA and TPA. The main component of the labeled tracer has an isoelectric point of 4.4 but there is some impurity in the tracer as it was shown in chromatofocusing.

Diagnostic tools and prognostic factors in human breast cancer evaluated by morphological and immunohistochemical methods

With regard to the clinical management of breast cancer besides the radiological (mammography and magnetic resonance), ultrasound and galactographic image processing systems we need safe and fast-working, histopathological and immunohistochemical methods to make in uncertain cases the diagnosis sure and to obtain well defined informations with respect to the individual biological behaviour of every single cancer to be treated under optimal conditions according to the specific personal situation (Fig.l).