Anti-miR-15a/16-1 has the ability to efficiently and specifically silence endogenous miR-15a and miR-16-1. Our data showed anti-miR-15a/16-1 could partly reverse the expression of WT1 in curcumin-treated K562 and HL-60 cells. These results

suggest that the decrease of WT1 expression is partly attributable to the increased expression of miR-15a and miR-16-1 in curcumin-treated leukemic cells. Thus our data suggest that one of the important anti-proliferation effects of curcumin on leukemic cells is via miRNAs pathway. Given that many miRNAs are regulated by pure curcumin, many further experiments will be required to define other miRNAs besides miR-15a/16-1 are regulated by curcumin and play an important role in anti-tumor effects www.selleckchem.com/products/cftrinh-172.html of curcumin. Conclusion Therefore, we conclude that pure curcumin can decrease WT1 expression partly through upregulating the expression of miR-15a and miR-16-1. Our data show for the first time that miRNAs pathway plays an important role in the function of anti-proliferation by pure curcumin in leukemic cells. Conflict of interests The authors declare that they have no competing interests. Acknowledgements The project supported by National Natural Science Foundation of China

(81172613), Zhejiang Provincial Natural Science Foundation of China (Y2101069, Y206383, Y12H080019), Scientifical Research Foundation (Y201119952) of Zhejiang Provincial Education Department. Electronic supplementary material Additional file 1: Figure S1. (A) K562 cells were treated with 5, 10, 20 uM pure curcumin for 48

hours, then PRT062607 manufacturer the mRNA level of WT1 was detected by qRT-PCR. ABL and GAPDH served as different housekeeping for normalization. (B) Primary leukemic cells of 12 AML patients were separated by Ficoll and were treated with 20 uM pure curcumin for 48 hours, then the mRNA levels of WT1 were detected by qRT-PCR. (C) The protein level of WT1 was detected by Western blotting after negative control(N.C), miR-15a and miR-16-1 mimics were transfected PtdIns(3,4)P2 into K562 for 48 hours. Figure S2. An illustration of the potential mechanisms of curcumin action in leukemic cells. Curcumin upregulated the expression of miR-15a/16-1 in leukemic cells. Overexpression of miR-15a/16-1 obviously reduced the protein level of WT1. However, downregulation of WT1 by siRNA could not increase the expression of miR-15a/16-1. These events showed that curcumin induced-upregulation of miR-15a/16-1 was an event upstream to the downregulation of WT1. Finally anti-miR-15a/16-1 oligonucleotides (AMO) partly reversed the down-regulation of WT1 induced by curcumin in leukemic cells and reversed the inhibition of cell proliferation caused by curcumin in K562 and HL-60 cells. (DOC 126 KB) ATR inhibitor References 1. Kreidberg JA, Sariola H, Loring JM, Maeda M, Pelletier J, Housman D, Jaenisch R: WT-1 is required for early kidney development. Cell 1993, 74:679–691.PubMedCrossRef 2.