Protein tyrosine kinase inhibitor, genistein, enhances apoptosis and cell cycle arrest in K562 cells treated with gamma-irradiation.

Genistein, is a natural isoflavone compound with a potent activity against protein tyrosine kinases. The leukemic cell line, K562, is a bcr/abl (Philadelphia chromosome) positive cell line that is resistant to DNA-damaging agents, including gamma-irradiation. Treatment with genistein increased apoptosis and promoted G2-phase arrest in the non-apoptotic population of the gamma-irradiated K562 cells. Irradiated cells that survived 72 h after the irradiation had a normal distribution in cell cycle, whilst genistein treatment kept cells arrested in the G2-phase, decreased the S-phase fraction and suppressed DNA-synthesis. Taken together, our results show that genistein augments apoptotic cell death after gamma-irradiation in K562 cells and this result cannot be attributed to abrogation of the G2/M checkpoint.

Antineoplastic and cytogenetic effects of complexes of Pd (II) with 4N-substituted derivatives of 2-acetyl-pyridine-thiosemicarbazone.

The effect of novel Pd(II) complexes with derivatives of 2-acetyl-pyridinethisemicarbazone, N4-ethyl (HAc4Et) and 3-hexamethyleneiminylthiosemicarbazone (HAchexim), on Sister Chromatid Exchange (SCE) rates and human lymphocyte proliferation kinetics was studied. Also, the effect of Pd(II) complexes on DNA synthesis of P388 and L1210 cell cultures and against Leukemia P388 was investigated. Among these compounds, the compound Bis(3-hexamethyleneiminyl-2-acetylpyridine-thisemicarbazonato++ +) palladium (II) was found to be distinctly effective against Leukemia P388, in inhibiting incorporation of 3H-thymidine into DNA and in inducing SCEs and cell division delays.

Comparative study of SCE induction and cytostatic effects by homo-azasteroidal esters of N,N-bis(2-chloroethyl)aminobenzoic acid in human lymphocytes.

The effect of homo-azasteroidal esters of benzoic acid mustard isomers and the 4-methyl derivatives, which have steroidal lactams as a biological basis, on cytogenetic damage was studied. Twenty compounds were comparatively studied, on a molar basis, as regards their ability to induce sister-chromatid exchanges (SCEs) and cell division delays. A correlation between potency for SCE induction, effectiveness in cell division delay and previously established antitumor activity of these compounds was observed.

Comparative study on cytogenetic damage induced by homo-aza-steroidal esters in human lymphocytes.

The effect of P[N,N-bis(2-chloroethyl)amino]phenylacetate esters of 3 beta-hydroxy-N-methyl-17 alpha-aza-D-homo-5 alpha-androstan-17-one (compound 3) and 3 beta-hydroxy-17 alpha-aza-D-homo-5 alpha-androstane (compound 2) on sister-chromatid exchange (SCE) frequencies and on human lymphocytes proliferation kinetics was studied. The results are compared with those of the P[N,N-bis(2-chloroethyl)amino]phenylacetate esters of 3 beta-hydroxy-17 alpha-aza-D-homo-5 alpha-androstan-17-one (compound 1). All compounds were found to be active in inducing markedly increased SCE rates and cell division delays. A correlation between potency for SCE induction, effectiveness in cell division delay and previously established antitumour activity of these compounds was observed.

Effects of tamoxifen and CV 205502 on the morphology and the evolution of the noncancerous mouse mammary gland.

Tamoxifen (TAM, 0.01 mg/animal, three times a week) and the experimental prolactin-lowering CV 205502 (CV, 1 microgram/animal, daily) were administered prophylactically, alone or combined, to virgin C3H/Sy mice during the early period of promotion in this spontaneous mammary carcinogenesis system (end of 2nd-5th month of age), in order to study their influence on the morphology and evolution of the noncancerous mammary gland during therapy and after treatment cessation. During TAM administration the epithelial cells of the growing part of the gland exhibited myoepithelial- and, late in the treatment period, apoptotic-like features instead of the secretory ones expected, accompanied by intense basement membrane alterations, thickening of the surrounding connective tissue and arrested adipocyte maturation. These effects reversed progressively after drug withdrawal. The epithelial alterations were more intense and longer lasting in the TAM+CV-group, while growth arrest of the glands was observed in both groups parallel to the degree and the duration of these morphological changes. In these groups, tumor incidence was diminished, as expected, but the tumors that developed late after treatment cessation were of low histological differentiation. The above morphological observations show that TAM inhibits noncancerous mammary gland growth during the reproductive period by altering stromal and epithelial differentiation, effects that reverse progressively after treatment discontinuation and are potentiated by a prolactin-lowering agent in this animal study.

Comparative study on cytogenetic effects by diplatinum complexes of the ligands of naphthazarine and squaric acid in human lymphocytes.

The effect of diplatinum complexes of the binucleating ligands of naphthazarine and squaric acid on Sister Chromatid Exchange (SCE) rates and human lymphocyte proliferation kinetics was studied. Squarodicisplatinum complex I, naphthazarindicisplatinum and squarodicisplatinum complex II induce cytotoxic effects as can be deduced from the resulted induction of SCEs and the produced cell division delays. Squarodicisplatinum complex I was found to be on a molar basis the most effective in causing markedly increased SCE rates and cell division delays. Cis-diaminodichloride platinum was found to be next in order of effectiveness with naphthazarindicisplatinum and squarodicisplatinum complex II following. Naphthazarine and SQA were found to be ineffective on induction of SCEs.

Induction of cytogenetic damage by modified steroidal derivatives of p-bis(2-chloroethyl)aminophenylacetic acid in human lymphocytes.

The effect of modified steroids, containing alkylating agents, on SCE rates and on cell kinetics in cultured human lymphocytes was studied. The homo-aza-steroidal ester of p-bis(2-chloroethyl)aminophenylacetic acid (ASE) was found to be the most effective in causing markedly increased SCE rates and cell division delays. The androsterone ester of p-bis(2-chloroethyl)aminophenylacetic acid (AE-CAPA) was found to be next in order of effectiveness with the lactone ester (LE-CAPA), chlorambucil ester 3 beta-hydroxy-13a-amino-13,17-seco-5a-androstan-17-oic-13,17-lactam (CBC-HAAL) and chlorambucil (CBC) following. p-Bis(2-chloroethyl)aminophenylacetic acid (CAPA) had only a small effect and 3 beta-hydroxy-13a-amino-13,17-seco-5a-androstan-17-oic-13,17-lactam (HAAL) had no effect at all. A correlation between potency for SCE induction, effectiveness in cell division delay and previously established antitumor activity of these drugs was observed.

Potential antitumor agents: steroidal amidoesters with an alkylating moiety.

17 beta-Acetamido-5-androsten-3 beta-ol-p-bis(2-chloroethyl) aminophenylacetate and 17 beta-acetamido-5 alpha-androstan-3 beta-ol-p-bis(2-chloroethyl)aminophenylacetate, amido steroidal alkylating agents, were investigated in L1210 and P388 leukemias, Ehrlich ascites tumor, and adenocarcinoma CA-755. Both substances were determined to increase the life span of the tumor-bearing animals and to cause several long-term survivors in adenocarcinoma CA-755.

Further studies on the antineoplastic activity of homo-aza-steroidal esters.

The modified steroidal alkylating agents, 17 beta-hydroxy-3-aza-A-homo-4 alpha-androsten-4- one(p-[bis(2-chloroethyl)amino]phenyl)butyrate(1),3 alpha-hydroxy- 13,17-seco-5 alpha- butyrate(2),3 beta-hydroxy-13,17-seco-5-androsten-17-oic- 13,17-lactam(p-[bis-(2-chloroethyl)amino]phenyl)butyrate(3) and and (p-[bis(2-chloroethyl)amino]phenyl)butyric acid(4) have been tested against L1210, P388, Ehrlich ascites tumors (EAT), Lewis lung (LL) carcinoma and adenocarcinoma CA-755. Of four compounds evaluated in L1210 leukemia, none displayed antileukemic activity. Almost all of the four compounds were more or less active against P388 leukemia. Compound 2 possesses a slight antitumor activity in EAT, while only compound 1 appears to be active in LL carcinoma. The antitumor activity of the three modified steroidal esters on adenocarcinoma CA-755 seems to be interesting.

Sialyltransferase and nucleoside diphosphatase as markers for tumor monitoring.

In this study we report serum sialyltransferase and nucleoside diphosphatase activities of patients with malignant tumors of various primary sites and extent, prior to and during chemotherapy. Enzyme levels were compared to clinical and laboratory parameters. The sialyltransferase and uridine diphosphatase (UDPase) activities in samples of 43 patients with advanced ovarian cancer was four to ten fold above the normal mean value (sialyltransferase 85.1 +/- 58 pmol/hr/ml and UDPase 26.6 +/- 7.2 nmol/hr/ml). After effective chemotherapy with adriamycin and cisplatin, the enzyme activity decreased markedly. In cases of complete remission, enzyme activity decreased to the normal range. In three cases after initial response for several months a rise of both enzymes was observed before any other biochemical finding of the forthcoming relapse. Similar patterns were observed in testicular cancer (6 cases). Clinical correlation is also obvious in other tumors except malignant lymphomas. Our findings show that the activities of these enzymes correlated with the clinical course, and therefore they can be the basis for clinical application for tumor monitoring, especially during chemotherapy.

Studies on the effect of homo-aza-steroidal ester of p-bis (2-chloroethyl) amino phenyl acetic acid in Ehrlich ascites tumor cells.

The effect of a homo-aza-steroidal ester (ASE) on the incorporation of radioactive precursor to DNA of Ehrlich ascites tumor (EAT) cells has been investigated. We found that treatment of cells with 80 micrograms/ml of ASE for 2 hours causes an inhibition of the incorporation of 3H-thymidine to DNA by 71%. This is partly because ASE affects the radioactive thymidine pool in the cell. The DNA from EAT cells after centrifugation in CsCl is shifted to higher densities when ASE is present throughout the experiment. This density shift was not observed when ASE was incubated only in the growth medium of the cells.

Interaction of homo-aza-steroidal ester of [p-[bis-(2-chloroethyl) amino]phenyl]acetic acid (ASE) with DNA of Ehrlich ascites tumor cells.

A cytostatic, homo-aza-steroidal ester of [p-[bis-(2-chloroethyl) amino]phenyl]acetic acid (ASE) was reduced with NaB3H4 and [3H]ASE-treated DNA prepared in vitro. We found that: (1) ASE reacts preferentially with purines; (2) ASE decreases the thermal stability of the double helix upon binding to DNA; (3) [3H]ASE binding sites are clustered along the DNA molecules; (4) ASE binding sites probably represent oligo- or polypurine sequences.

[Chemotherapy of the non-small cell bronchogenic carcinoma]. / Chemotherapie des nicht-kleinzelligen Bronchialkarzinoms.

From 1976 to 1981 61 patients (50 men and 11 women) with non-small cell bronchogenic carcinoma (i.e. squamous cell or adeno-carcinoma) received a chemotherapy treatment based on two cytostatic drug combinations for each histological group. The drugs which were used were cyclophosphamide, adriamycin, vincristine, methotrexate, bleomycin, 5-fluorouracil and procarbazine. All patients had advanced disease and 6 of them had had previous surgical excision of the primary tumor. A large proportion of the patients had received irradiation to the primary site prior to chemotherapy. 1 patient developed a complete remission, 5 developed a partial remission and 34 showed stabilization of their disease. In 21 patients the disease progressed in spite of the treatment. Responders had a significantly longer survival than non-responding patients.

Hodgkin's disease in Greece: a retrospective clinicopathologic study of 78 cases.

The Rye histologic classification of Hodgkin's disease has been applied retrospectively to a series of 78 previously untreated cases of Hodgkin's disease. The patients have been followed for periods varying from 1 to 12 years. Mixed cellularity was the most common histologic type. Nodular sclerosis had an intermediate frequency, and lymphocytic predominance and lymphocytic depletion showed the lowest incidence. The sex incidence showed a male predominance in all histologic types except the nodular sclerosis, where prevalence of females was noted. The study confirms the better prognosis of the lymphocytic predominance and nodular sclerosis in comparison with the poor prognosis of the mixed cellularity and the very poor prognosis of the lymphocytic depletion. Nodular sclerosis presented with higher incidence in the mediastinum; it occurred mainly in young females and it exhibited a relatively good prognosis, findings which support the accepted view that nodular sclerosis comprises a distinct clinical and biological type of the disease. A marked difference in the survival rates in Stages I and II as compared to those of Stages III and IV was found.

New compounds: antitumor activity of 3beta-hydroxy-13alpha-amino-13,17-seco-5alpha-androstan-17-oic-13,17-lactam 4-[p-[bis(2-chloroethyl)amino]phenyl]butyrate.

3beta-Hydroxy-13alpha-amino-13,17-seco-5alpha-androstan-17-oic-13,17-lactam 4-[p[bis(2chloroethyl)amino]phenyl]butyrate was prepared by reacting 4-[p-[bis(2-chloroethyl)amino]phenyl]butyryl chloride hydrochloride with 3beta-hydroxy-13alpha-amino-13,17-seco-5alpha-androstan-17-oic-13,17-lactam. They cytostatic action of the ester was investigated on two tumor systems (B16 melanoma on C57 b1 mice and T8-Guerin on rats).

DNA-dependent RNA polymerase from T8 Guerin tumor.

DNA-dependent RNA polymerases from nuclei of T8 Guerin tumor were studied. Two enzymes were purified several hundred times by the use of ammonium sulfate precipitation, DEAE-cellulose and phosphocellulose chromatography. One of them belongs to A(I) RNA polymerases and the second to B(II) as was established from their metal and ionic strength requirements. activity in the presence of native and denatured DNA and the resistance to a-amanitin inhibition. The quantity of class A enzyme was increased compared to B, a fact observed with most neoplastic tissues so far studied. This increase of the polymerase responsible for ribosomal RNA synthesis could probably be related to malignant transformation in animals.

Note on the ultrastructure of the Guérin T-8 transplantable tumour.

The ultrastructure of the Guérin tumour T-8 is described. Although this transplantable tumour has its ancestry in an uterine adenocarcinoma, there is no more evidence of gland formation. Desmosomes, or any other form of cell attachment organelle, were totally absent. The tumour has now a very anaplastic sarcomatous appearance.