Modification of membrane fluidity and depolarization by some anthracyclines in different cell lines.

One of the major problems in cancer treatment is the progressive desensitization of cancer cells to chemotherapeutic drugs. Several hypotheses have been advanced to explain this property of cancer cells. In recent years different calcium channel blockers and other chemosensitizing agents like synthetic progestins have been used to revert drug resistance. In our experiments we evaluated the effects of Doxorubicin and Idarubicin on membrane fluidity and depolarization using normal lymphocytes, chronic lymphatic leukemia lymphocytes, normal breast and hormone dependent breast cancer cells and cardiomyocytes. The drugs were used alone or in combination with Verapamil and Medroxyprogesterone acetate. We showed that MPA enhances DOXO and IDA biochemical effects, acting not only on the membrane lipid bilayer, but also on ion channels. VERA instead does not seem to act through the same mechanism.

The biological approach to radioimmunoscintigraphy.

The authors have analysed the theoretical and clinical implications of a biological approach to radioimmunoscintigraphy (RIS), a new diagnostic technique based on the in vivo reaction between radiolabelled monoclonal antibodies and tumor-associated antigens expressed by the neoplastic cells. The assumption is that, as in all radioisotopic procedures, the radioimmunoscintigraphic image is the expression of differences in concentration (and not in density) between adjacent tissues. This "biological premise" is analysed to demonstrate a possible role of RIS not only in diagnosis but also in prognosis and therapy. A comparison with other imaging procedures and with histopathology, considered as the "gold standard" of the present-day morphological approach to diagnosis, is also discussed. Finally, biological premises, as well as the initial results and prospects regarding the use of high doses of radiolabelled monoclonal antibodies for therapeutic purposes are presented.

Inhibition of cyclophosphamide mutagenicity by beta-carotene.

Cyclophosphamide (CP) metabolites, rather than the parent compound, show mutagenic activity towards Salmonella typhimurium TA 1535 tester strain when S9 fraction from phenobarbital (PB)-induced rat liver is used as in vitro metabolizing system. On the other hand, inhibition of CP in vitro mutagenicity was observed by adding increasing amounts of beta-carotene (beta-C) to the system. A typical dose-dependent mutagenic response was observed by assaying 24 h urine samples of PB-induced rats injected i.p. with different amounts of CP. Addition of beta-C to urines of CP-treated rats failed to inhibit their mutagenicity. Conversely, a marked decrease in urine mutagenicity was observed when rats were simultaneously treated with the two drugs. These data show that beta-carotene partially inhibits, in vitro and in vivo, CP metabolism via hepatic mixed-function oxidase enzymes to mutagenic species.

Intermediate and high grade malignant non-Hodgkin's lymphomas: preliminary results using a new combination regimen (EVE-COPEM).

Between 1988 and 1992, 60 patients with intermediate and high-grade non-Hodgkin's lymphomas (NHL) were treated with a new multidrug combination chemotherapy including 4'epidoxorubicin (25 mg/m2), etoposide (60 mg/m2), cyclophosphamide (400 mg/m2), administered intravenously (i.v.) on the 1st, 2nd and 3rd day every 4 weeks, prednisone (40 mg/m2) orally for 6 days every 4 weeks, vincristine (1 mg/m2) i.v. and methotrexate (400 mg/m2) i.v. on the 8th day every 4 weeks, vindesine (2.5 mg/m2) and cytarabine (200 mg/m2) on the 15th day every 4 weeks. Patients achieving apparent complete remission (CR) or good partial response (PR) after the 1st cycle of therapy were submitted to three other cycles of the same therapy. Patients failing to respond to the 1st cycle or whose disease progressed despite therapy, were treated with an alternative 2nd line therapy. Seventeen patients (28%) had stage II-II E, 15 (25%) stage III and 28 (47%) stage IV disease. Tumoral mass > 10 cm was found in 28 cases, the presence of extranodal sites (ES) in 32 cases, serum lactate dehydrogenase (LDH) > 240 IU/l in 34 cases, performance status (PS) > or = 2 in 12 cases. CR was obtained in 46 (76.4%) out of the 60 patients. Relapse-free survival (RFS) was 82, 64 and 61% with a median follow-up of 12, 24 and 36 months respectively. No relapse occurred later than 26 months after achievement with CR thus far. Overall survival (OS) was 77% at 12 months and calculated to be 62% and 59% at 24 and 36 months, respectively. Two patients died as a result of the treatment. Reversible myelosuppression was the main toxic effect. One hundred and ten out of the 221 cycles of chemotherapy were delayed because of therapy toxicity. Negative prognostic factors on the RFS and OS were the presence of an advanced stage of disease, a mass larger than 10 cm, the presence of ES, the elevated LDH, the PS > or = 2, the delay of therapy. In conclusion, results obtained using our protocol overlap those from other third generation regimens. Toxicity was also similar. The influence of clinical conditions such as stage of disease, the presence of ES, high LDH level and tumoral mass > 10 cm on the RFS and OS were significant. Principal variables influencing prognosis must be unified to compare results of similar treatments from different institutions.

Management of patients with ovarian cancer using monoclonal antibodies.

We describe in detail the current trend using monoclonal antibodies to diagnose ovarian cancer either in vitro or in vivo. The approach with such powerful reagents allows to differentiate in vitro tumor histotypes and to detect in peritoneal washings the presence of a few neoplastic cells which characterize the minimal disease. The detection of elevated sera levels of ovarian cancer-associated antigens, such as CA-125 and TAG-72, allows the monitoring, follow-up of these patients and the response to therapy with great accuracy. We focused our attention on the role in vivo of labelled monoclonal antibodies, mainly for diagnostic purposes. Radioimmunoscintigraphy has been found to be more reliable than CT and US to detect foci of disease mainly in patients already treated by surgery, overcoming all the problems usually encountered with these two procedures.

A prospective imaging study of 131I-B72.3 monoclonal antibody in patients with epithelial ovarian cancer: preliminary report.

Twenty patients with known ovarian cancer have been investigated in this pilot study to verify the clinical usefulness of radioimaging using the B72.3 monoclonal antibody labelled with iodine-131. No adverse reactions occurred after intravenous injection of B72.3 MoAb. The radioimaging results were compared with those obtained with other diagnostic methods, including computed X-ray tomography and ultrasound. A sensitivity of 85% in the detection of primary ovarian cancers and collections of ascites, and of 84% in the detection of abdominal and extraperitoneal metastases has been demonstrated using this radioiodinated antibody in vivo. No false localization occurred. Immunohistochemical studies showed no cross-reactions between B72.3 MoAb and mesothelial cells, confirming the high specificity of binding between B72.3 MoAb and neoplastic cells in ascites. Negligible uptake of radiolabelled B72.3 MoAb has been demonstrated in the unaffected ovary. The major advantage of using this monoclonal antibody is related to the expression of a recognized antigen (called TAG 72) in mucinous, serous and in differentiated adenocarcinomas of the ovary.

[Mechanism of action of progestins in the treatment of hormone-dependent breast cancer (author's transl)]. / Sul meccanismo d'azione dei progestinici nella terapia dei tumori mammari ormono - dipendenti

The in vitro interference of some gestagens with the binding of 3H-17-beta-oestradiol to cytosol specific receptors was investigated with a view to elucidating the mechanism of action of progestins in the treatment of human hormone-dependent breast cancer. A decrease (up to 85%) of oestradiol binding capacity was observed with high concentrations of progesterone, clogestone and medrogestone. These findings are in good agreement with those previously obtained by the same progestins in our laboratory on rat uterine estrogen receptors in vitro or in vivo. These results provide the support for the hypothesis that the mode of action of progestins in the therapy of mammary and perhaps uterine carcinomas is to some extent related to the inhibition of oestradiol binding to cytosol specific receptors.

PIP: For the purpose of explaining the mechanism of action of progestins in the treatment of human hormone-dependent breast cancer, interference of some progestins with the binding of radioactive 17beta-estradiol to its specific receptors in a cytosol substrate in vitro was studied. A decrease of up to 85% in the binding capacity of estradiol was observed with high concentrations of progesterone, clogestone, and medrogestone. These findings appear to confirm those obtained by the same authors using the same progestins on rat uterine estrogen receptors in vitro and in vivo, and support the hypothesis that the mechanism of action of progestins in the treatment of breast and possibly uterine cardinoma is to some extent related to the inhibition of estradiol binding to its specific receptors, thereby inhibiting the formation of the estrogen-receptor system which is the cause of cell growth in such tumors. This would also explain the greater frequency of successful treatment when using higher doses of progestins.

A phase II study of paclitaxel/cisplatin combination in patients with metastatic breast cancer refractory to anthracycline-based chemotherapy.

AIMS AND BACKGROUND: To investigate the safety and efficacy of a paclitaxel and cisplatin regimen in a selected group of metastatic breast cancer patients with primary or acquired chemo-resistance to anthracycline-based chemotherapy. PATIENTS AND METHODS: Thirty-eight consecutive women with metastatic breast cancer (PS < or =2) were entered in this phase II trial; all patients had been previously treated for metastatic disease with chemotherapy containing anthracyclines and had shown a progression of the disease during drug administration or after a clinical response lasting less than 6 months. Fifteen patients had received 2 or more chemotherapeutic regimens for advanced disease; 31 patients had > or =2 sites of metastatic disease. Paclitaxel (135 mg/m2) was administered iv by a 3-hr infusion followed by iv infusion of cisplatin (75 mg/m2) on day 1, every 3 weeks for 6 cycles. After the completion of the planned chemotherapy administration, 9 responsive patients continued to receive paclitaxel alone (175 mg/m2) iv, on day 1, every 3 weeks, until disease progression or unacceptable toxicity. RESULTS: A partial clinical response was recorded in 17 cases (45%; 95% CI, 30-64%). The median duration of overall response was 8 months; for the 9 responsive patients who continued treatment with paclitaxel alone, 4 had maintained the partial clinical response at the median follow-up of 24 months from the onset of therapy. The median time to progression was 6 months and median overall survival 8 months. Neurotoxicity was the most frequent adverse effect and caused treatment discontinuation in 5 cases for grade 3-4 paresthesia and/or an arthralgia/myalgia syndrome. Grade 3-4 neutropenia occurred in 16 patients (44%). CONCLUSIONS: Paclitaxel/cisplatin is an active regimen for the treatment of patients with metastatic breast cancer refractory to anthracycline-based chemotherapy. However, the cumulative neurotoxicity should limit the efficacy of prolonged paclitaxel monotherapy in responsive patients.

Effect of beta-carotene on mutagenic activity of some antineoplastics.

Mutagenic activity on Salmonella typhimurium strains of some cytostatic drugs in the absence and in the presence of beta-carotene was evaluated. Cyclophosphamide mutagenicity was reduced by the retinoid both in vitro and in vivo. Conversely, its metabolite 4'-hydroxy-cyclophosphamide, which does not require enzymatic transformation to exert genotoxic activity against bacteria, was not affected by the presence of beta-carotene. Moreover, the mutagenic activity of cis-Platinum, Adriamycin and 4'Epiadriamycin, which are typical direct-acting mutagens, was not affected by beta-carotene. Data obtained confirm that beta-carotene is able to prevent mutagenic activity of cyclophosphamide by interfering with its metabolic activation but failed to inhibit the interaction of genotoxic compounds with bacterial DNA.

Lonidamine alone and in combination with other chemotherapeutic agents in the treatment of cancer patients.

The tolerance and antitumor activity of Lonidamine administered alone and in combination with other anticancer agents were studied. Myalgia was the most frequent side effect; there were no changes in the hematological parameters attributable to Lonidamine administration. 1 partial response out of the 16 patients treated with Lonidamine alone was observed.

Is there any impact of new drugs on the outcome of advanced NSCLC? An overview of the Southern Italy Cooperative Oncology Group trials.

Lung cancer represents the major cause of cancer-related death in Europe and North America, accounting for 28% of all cancer deaths. Seventy to 80% of all lung cancers are non-small cell lung cancers (NSCLCs), and approximately 75 % of these patients present with locally advanced or disseminated disease. Even though chemotherapy is now recommended in the majority of cases of unresectable NSCLC, it still fails to substantially modify the fate of these patients. In recent years, several active cytotoxic drugs (paclitaxel, docetaxel, vinorelbine, gemcitabine, and irinotecan) have been developed, showing an overall response rate (ORR) <20% in NSCLC. Phase II/III trials testing these new agents in combination with cisplatin have been carried out in recent years with inconsistent results. Large randomized trials testing cisplatin-paclitaxel, carboplatin-paclitaxel, and cisplatin-gemcitabine regimens have been reported showing no substantial superiority of these combinations over standard treatments. The ORR remained well below 50%, and the median survival times were quite far from one year. These data could suggest that the addition of a single new agent to a platinum compound could be insufficient to substantially improve the prognosis of advanced NSCLC patients. In view of these disappointing data, the Southern Italy Cooperative Oncology Group has tried to improve the fate of patients with advanced NSCLC by testing new triplet combinations, which combined cisplatin with two rather than one of the newest chemotherapy agents. To avoid an unacceptable increase in toxicity and/or a marked decrease in dose intensity, the standard schedules of administration of the three agents used in these studies were changed, and the schedule changes were evaluated in phase I trials aimed at determining the maximum tolerated dosages of the drugs. Subsequently, phase II and III trials were conducted. The present paper summarizes the results of the clinical trials either completed or under way and aims to evaluate whether this strategy will result in a substantial prognostic improvement.

(131I)meta-iodobenzylguanidine scintigraphy and selective venous catheterization after thyroidectomy for medullary thyroid carcinoma.

Fifteen patients with medullary carcinoma of the thyroid (MCT), who had persistently elevated levels of serum calcitonin (CT) and carcinoembryonic antigen (CEA) after total thyroidectomy, were studied in order to localize the sites of the recurrent disease. Routine diagnostic examinations, including ultrasonography (US) and computed axial tomography (CAT), were carried out in all the cases. Scintigraphy with radio-iodinated metaiodobenzylguanidine ((131I)-MIBG) was performed in 13 cases; selective venous catheterization (SVC) to reveal a gradient of CT levels was performed in 12 cases. Ten patients underwent both (131I)-MIBG scintigraphy and SVC. US and CAT revealed the sites of recurrent tumor in only 4 out of the total 15 patients. SVC in basal conditions showed the presence of small metastases in 2 cases, and after intravenous stimulus with pentagastrin in 4 others. The MIBG scan showed metastatic foci of sporadic MCT in 2 patients, residual medullary thyroid tissue in 4 others, and a pheochromocytoma in a previously undiagnosed patient with Sipple's syndrome. More particularly, MIBG scan and SVC showed the localization of residual or metastatic tumor in 10 cases. In all 10 cases, results of the MIBG scan and SVC were confirmed as true positive by subsequent surgery and histopathologic examination.(ABSTRACT TRUNCATED AT 250 WORDS)

High serum thyroglobulin levels. Diagnostic indicators in patients with metastases from unknown primary sites.

Patients with metastases from differentiated thyroid carcinoma have a good chance of long-term survival when the diagnosis is prompt and appropriate therapy is applied early. This is also true for patients with metastases in the bone, taking into account that an appropriate therapy, usually 131-I, may be palliative in some cases. This study investigates whether serum thyroglobulin (Tg) measurement in patients with cold thyroid nodule and metastases from an unknown primary site could help identify a differentiated thyroid carcinoma. The results obtained show that Tg measurements is a useful adjunctive test when used with fine-needle aspiration biopsy. In particular, very high Tg levels point to metastatic thyroid cancer, whereas lower levels do not help determining whether metastatic cancer is of thyroid origin or not.

Diagnosis of ovarian cancer with radiolabelled monoclonal antibodies: our experience using 131I-B72.3.

Radioimmunoscintigraphy (RIS) using 131I-labelled B72.3, a monoclonal antibody (MoAb) reacting against a tumor associated antigen (TAA) called TAG-72, has been performed in 36 patients with epithelial ovarian cancer. The patients were divided in three groups as follows: 17 patients with primary cancer before any therapy (Group 1); 10 patients studied after a partial therapeutic approach, having either bulky or minimal disease (Group 2); 9 patients with microscopic disease or in clinical remission at the moment of the study (Group 3). All the most important epithelial histotypes, including mucinous, were present. Results were confirmed at surgery and/or by other diagnostic procedures. Immunocytochemical (ICC) and immunocytofluorimetric (ICF) studies on ascitic collections were performed in order to demonstrate specificity of B72.3 and TAG-72 distribution on neoplastic cells. Immunohistochemistry (IHC) on tissue sections was also obtained. No cross reactions between B72.3 and mesothelial cells in the presence of specific uptake by neoplastic cells was found. Moreover, a non-homogeneous distribution of TAG-72 in the neoplastic population was demonstrated by ICF. RIS proved the intraperitoneal presence of disease in 15 out of 17 and in 5 out of 10 patients in Groups 1 and 2, respectively. One out of four (Group 1) and two out of four (Group 2) extraperitoneal metastases were also seen. False negative results were explained by lack of expression of the antigen, size and location of the lesion, and patho-physiological conditions. One false positive due to an aspecific uptake by a post-surgical active scar was also observed in a disease-free patient.

Cancer antigen 125, tissue polypeptide antigen, carcinoembryonic antigen, and beta-chain human chorionic gonadotropin as serum markers of epithelial ovarian carcinoma.

Cancer antigen 125 (CA 125) is common to most epithelial ovarian tumors. Therefore, it is potentially a good marker of this disease. This hypothesis was evaluated by measuring the serum levels of CA 125 in 81 patients with ovarian cancer (25 with nonactive and 56 with active disease), in 105 patients of both sexes with nonovarian tumors, and in 171 healthy controls of both sexes. The serum levels of three other markers, tissue polypeptide antigen (TPA), carcinoembryonic antigen (CEA), and human chorionic gonadotropin, beta subunit (beta-hCG), were also measured in the same 357 subjects. The results of this study clearly indicate the clinical irrelevance of both CEA and beta-hCG as tumor markers in ovarian carcinomas. Conversely, the clinical usefulness of CA 125 and TPA was confirmed. In particular, CA 125 and TPA showed comparable sensitivity, while CA 125 showed a higher specificity for ovarian cancer than TPA. The association of CA 125 with TPA was very useful in continuous observation of patients with active disease in order to evaluate the clinical effectiveness of the therapy. Moreover, for patients in clinical remission, the markers allowed early detection of a recurrence of the disease.

Laevofolinic acid, 5-fluorouracil, cyclophosphamide and escalating doses of epirubicin with granulocyte colony-stimulating factor support in locally advanced and/or metastatic breast carcinoma: a phase I-II study of the Southern Italy Oncology Group (GOIM).

Sixty-four consecutive patients with locally advanced (n = 7) or metastatic breast cancer (n = 57), were treated with a combination of laevofolinic acid 100 mg m-2 plus 5-fluorouracil 340 mg m-2 i.v. on days 1-3, cyclophosphamide 600 mg m-2 i.v. on day 1 and epirubicin 90 mg m-2 i.v. on day 1. Epirubicin dose was progressively escalated by 10 mg m-2 per cycle up to 120 mg m-2 in the absence of dose-limiting toxicities. Granulocyte colony-stimulating factor (G-CSF) was given subcutaneously in order to prevent neutropenia. Epirubicin dosage could be increased to 100 mg m-2 in 53 patients (87%), to 110 mg m-2 in 31 patients (51%) and to 120 mg m-2 in 18 cases (30%). In most patients the dose-limiting toxicity was represented by myelosuppression. A statistically significant correlation was found between median white blood count (WBC) or absolute neutrophil count (ANC) nadir and epirubicin dose level (P = 0.009; P = 0.008). Moreover, a statistically significant correlation was observed between the number of chemotherapeutic cycles, nadir ANC and WBC and the occurrence of anaemia and thrombocytopenia of increasing severity. These data suggest the occurrence of progressive cumulative bone marrow toxicity. Although patients who reached different epirubicin levels showed differences in mean dose intensity, such differences were not statistically significant. No correlation was found between the increase in dose intensity and type, rate or duration of objective responses. In patients with metastatic breast cancer the overall response rate was 72% (95% CL 66-78%) with a 25% complete response rate. Median duration of response was 10 and 13 months respectively for complete and partial responses. All patients with locally advanced breast cancer had an objective response and underwent radical mastectomy. Projected median survival of the whole series of patients with metastatic breast cancer was 20 + months. These data demonstrate that the combination of 5-fluorouracil with laevofolinic acid, cyclophosphamide and epirubicin is very active against metastatic breast cancer. Use of G-CSF allows epirubicin dosage to be increased up to 120 mg m-2 cycle-1, but its use may be linked to the occurrence of sometimes severe cumulative haematological toxicity.

Clinical management of prolactinomas: a ten-year experience.

A ten-year experience on 36 patients bearing macroprolactinomas (MP) and 86 others bearing microprolactinomas (mP) is reported in this study. Different therapeutical approaches were used: 1) trans-sphenoidal surgery in 24 patients with MP and in 25 with mP; 2) medical therapy with the oral form of bromocriptine (BRC) in all the 24 patients with MP previously subjected to surgery, in 48 patients with mP ab initio, and in 16 out of 25 patients with mP previously subjected to surgery; 3) medical therapy with the long-acting injectable forms of BRC in 12 MP- and 13 mP-bearing patients, and 4) conventional radiotherapy in 12 of the 24 patients with MP previously subjected to surgery. The follow-up, performed five years after surgery, showed that: a) all the 24 patients with MP but one had normal PRL levels during BRC administration, with a rebound of hyperprolactinemia in all cases after withdrawal; b) during the treatment BRC caused normalization of PRL in 15 of the 16 mP-bearing patients surgically treated and in all the 48 mP-bearing patients only treated with BRC; c) in 20 of the 25 patients the treatment with injectable retard BRC caused the normalization of plasma PRL and the shrinkage of the tumor mass in all the patients with MP but one, as revealed by seriate CT scans. In conclusion, the surgical treatment of prolactinomas was ineffective to normalize plasma PRL levels in most patients whereas BRC, in standard or in retard forms, was able to normalize plasma PRL levels, reduce the tumoral mass and preserve the pituitary residual tissue. BRC should be, therefore, used as first choice therapy both for MP and mP.

Evaluation of TAG-72 as a serum marker in ovarian and breast carcinoma.

Tumor Associated Glycoprotein 72 (TAG-72) is common to most epithelial tumors. In this study serum levels of TAG-72 were measured in 36 healthy female subjects, in 94 patients with breast cancer, and in 43 others with epithelial ovarian cancer. More particularly, 27 out of the 94 patients with breast cancer had early disease (Stage I-I; or T 0-2b; N 0-1b; M 0), while the remaining 67 had advanced disease (Stage III-IV; or T 0-4; N 0-3; M 0-1). All the 43 subjects with ovarian cancer were at stage III-IV (FIGO Classification), 12 of whom had minimal disease (lesions less than 2 cm) and the other 31 had bulky disease. 3.5 U/ml being the highest value found in the control group, we arbitrarily assumed 3.85 U/ml (mean +/- 3 SD) as the cut-off limit in this preliminary study. Among the patients with breast cancer, 17 out of the 67 subjects with advanced disease (25.3%) and 1 out of the 27 others with early disease (3.7%) exceeded the TAG-72 cut-off limit. Among the patients with ovarian cancer, 2 out of the 12 subjects with minimal disease (16.7%) and 22 out of the others 31 with bulky disease (70.9%) had TAG-72 levels above cut-off limit. High TAG-72 levels were found in all the histotypes of ovarian cancer including the mucinous type. Preliminary data seem to indicate that in ovarian cancer variations in serum levels of TAG-72 are in agreement with the trend of the disease.(ABSTRACT TRUNCATED AT 250 WORDS)