Antiproliferative effect of indomethacin on CML cells is related to the suppression of STATs/Bcl-XL signal pathway.
Indomethacin (IN) can inhibit cyclooxygenase activity and is considered to exert antitumor action in a variety of cancer cells. In the present study, we investigated the underlying mechanism of its antiproliferative effect on chronic myeloid leukemia (CML) cells. We studied the role of signal transducer and activator of transcription 1 or 5 (STAT(1) or STAT(5)) and Bcl-X(L) proteins in IN-induced proliferative inhibition on CML cells. Both K562 cells and fresh bone marrow mononuclear cells from five CML patients were exposed to IN. Cell proliferation was determined by MTT assay. The expression of JAK(2), STAT(1), STAT(5), and Bcl-X(L) proteins was probed with Western blotting. The level of phosphorylated STAT(1) (p-STAT(1)) or STAT(5) (p-STAT(5)) proteins was determined by coimmunoprecipitation combined with Western blotting. Intracellular localizations of both STAT(1)/STAT(5) and p-STAT(1)/p-STAT(5) were observed by indirect immunofluorescence assay. Our results showed that IN could inhibit the proliferation of CML cells in a dose-dependent manner (36-288 microg/ml). The expression of STAT(1) and STAT(5) was suppressed by IN both in a concentration-dependent manner and a time-dependent (0-36 h) manner. The levels of p-STAT(1) and p-STAT(5) were down-regulated by IN. A similar result was obtained for Bcl-X(L) protein expression. The intracellular fluorescence signals representing STAT(1)/STAT(5) and p-STAT(1)/p-STAT(5) were obviously weakened by IN. In contrast with IN, granulocyte-macrophage colony-stimulating factor could significantly promote the growth of CML cells and up-regulate the expression of both STAT(1)/STAT(5) and p-STAT(1)/p-STAT(5). This data indicated that IN is able to suppress the proliferation of CML cells, and the mechanism is associated with the inhibition of STATs/ Bcl-X(L) signal transduction pathway.
Institute of Molecular Hematology, the Second Xiang-ya Hospital, Central South University, Changsha, Hunan, 410011, People's Republic of China.
Guangsen
G
Zhang
Yunbi
Y
Fu
D002455:Cell Division / Q000187:drug effects; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology*; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics*; D050791:STAT Transcription Factors / Q000037:antagonists & inhibitors*; D050794:STAT1 Transcription Factor / Q000037:antagonists & inhibitors; D015398:Signal Transduction / Q000187:drug effects*; D014407:Tumor Cells, Cultured; D051020:bcl-X Protein / Q000037:antagonists & inhibitors*
D050791:STAT Transcription Factors; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D051020:bcl-X Protein; D007213:Indomethacin