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We have shown previously that withaferin A (WA), a promising anticancer constituent of Ayurvedic medicine plant Withania somnifera, inhibits growth of human breast cancer cells in culture and in vivo in association with apoptosis induction. The present study builds on these observations and demonstrates that WA inhibits constitutive as well as interleukin-6 (IL-6)-inducible activation of signal transducer and activator of transcription 3 (STAT3), which is an oncogenic transcription factor activated in many human malignancies including breast cancer. The WA treatment (2 and 4 μM) decreased constitutive (MDA-MB-231) and/or IL-6-inducible (MDA-MB-231 and MCF-7) phosphorylation of STAT3 (Tyr705) and its upstream regulator Janus-activated kinase 2 (JAK2; Tyr1007/1008) in MDA-MB-231, which was accompanied by suppression of their protein levels especially at the higher concentration. Exposure of MDA-MB-231 or MCF-7 cells to WA also resulted in suppression of (i) transcriptional activity of STAT3 with or without IL-6 stimulation in both cells; (ii) dimerization of STAT3 (MDA-MB-231) and (iii) nuclear translocation of Tyr705-phosphorylated STAT3 in both cells. To our surprise, the IL-6-stimulation, either before or after WA treatment, did not have an appreciable effect on WA-mediated apoptosis in MDA-MB-231 or MCF-7 cell line. The IL-6-stimulated activation of STAT3 conferred a modest protection against WA-mediated suppression of MDA-MB-231 cell invasion. General implication of these findings is that WA can trigger apoptosis and largely inhibit cell migration/invasion of breast cancer cells even after IL-6-induced activation of STAT3, which should be viewed as a therapeutic advantage for this agent. [ABSTRACT FROM PUBLISHER]

In the present studies we have compared the levels of glutathione (GSH) and GSH-related enzymes in lung tumors and corresponding normal tissues obtained from the same individuals. We have also immunologically quantitated the relative amounts of glutathione S-transferase π (or GST-P) type antigen in tumors and adjacent normal tissues from five patients. GST activities towards 1-chloro-2, 4-dinitrobenzene (CDNB) and ethacrynic acid were found to be elevated in tumors from two out of five patients (patients #1 and 4), whereas the activity towards these substrates was markedly suppressed in the tumor tissue from one of the patients (#5). Immunotitration and Western blot studies using antibodies raised against π-type GST isoenzymes of human lung and placenta indicated induction of GST π-type isoenzyme in tumors from patients #1 and 4 and suppression of this isoenzyme in tumor from patient #5. The tumors from patients #2 and 3 did not show any increase in GST activity or GST π-type antigen. Except for the tumor from patient #5, the GSH content was higher in the tumors from other patients. GSH reductase activity was found to be elevated in tumors of all the patients examined in this study. These results indicate that GSH and GSH related enzymes are differentially altered in lung tumors and GSH levels and GST π- or GST-P-type isoenzyme(s) are not uniformly elevated in all tumors. [ABSTRACT FROM PUBLISHER]

A glutathione S-transferase (GST) isoenzyme having common antigenicity to rat placental form (GST-P) and human placental form (GST-π) has recently been suggested may be a marker of carcinogenesis. In the present study we have investigated the expression of this isoenzyme in three small cell lung cancer cell lines in order to determine whether or not this isoenzyme can be used as a general marker of carcinogenesis. GST activity towards 1-chloro-2,4-dinitrobenzene in two of the cell lines (NES and NOC-361) was moderately higher than that in normal human lung, but this activity was markedly suppressed in one of the cell lines (NCI-H69). Quantitation of the GST isoenzymes in the tumors grown in nude mice by injecting these cell lines also revealed a moderate increase of GST-π-type isoenzyme in NES and NOC-361 and its suppression in NCI-H69. Immunocytochemical localization studies with these tumors using antibodies raised against GST-π also indicated a drastic decrease of GST-π-type isoenzyme in NCI-H69 and this finding was confirmed by Western biot studies. These results suggest that GST-π or the isoenzyme(s) having similar immunological nature to GST-π, cannot be used as the general markers of neoplastic states. [ABSTRACT FROM PUBLISHER]