4 or 3.2 mM cinnamic acid for 6 (A, B, C), 12 (D, E, F) and 24 hours (G, H, I). The results did not show differences among the control groups and the treated groups. We did not observe significant differences between the control and treated groups after 6 or 12 hours of drug exposure (Table 3). Interestingly, the apoptotic cascade in the HT-144 cells was initiated approximately 24 hours after treatment with 3.2 mM cinnamic acid, specifically, when the frequency of cell death changed from 5% in the control group to 30% in the treated group. Our

results indicated that there was no significant increase in apoptotic cell frequency PCI-34051 after treatment with 0.4 mM of the drug. Table 3 Frequencies (%) of apoptotic cells (early + late apoptosis) in HT-144 and NGM cell lines after treatment with cinnamic acid in different times and concentrations Cell line Time of treatment Control groups Treated groups       0.05 mM 0.4 mM 3.2 mM HT-144 6 hours 7.48 6.96 5.74 6.45 12 hours 2.78 2.29 2.77

7.20 24 hours 4.51 4.52 buy Sapanisertib 3.16 29.53a NGM 6 hours 9.59 8.83 7.07 6.64 12 hours 4.44 4.46 2.97 2.92   24 hours 3.75 4.64 3.90 5.82 The results were obtained by quantification of cells positive to activated-caspase 09 by using a flow cytometer. a Significantly different from control group according to Multidimensional Nonlinear Descriptive Analysis. Furthermore, there were no differences between the control and treated groups of NGM cells after 24 hours of treatment with cinnamic acid (Table 3). The frequency of apoptotic cells TGF-beta inhibitor in the control group was approximately 5%, and the frequency of apoptosis in the NGM cell line did not reach 9% in any group. The statistics confirmed that the differences observed were not significant. The western blotting analysis showed that both cell lines

express the p53 protein. We could not confirm the selective effects of cinnamic acid by the total p53 quantification or p53 phosphorylation because apoptosis in HT-144 cells was not directly associated with the increase of p53 expression or phosphorylation (Figure 4). Figure 4 p53 and phospho-p53 levels in NGM and HT-144 cells after cinnamic acid exposure for 24 hours. There were no differences in p53 or phospho-p53 levels after treatment of NGM cells. HT-144 cells showed decreased level of p53 and phospho-p53 after treatment with cinnamic acid. Tubulin was used as a loading control. Cell morphology The morphological changes observed using microscopy after treatment with cinnamic acid and the BrdU incorporation data suggested that the drug targets the cell cycle. Thus, we analyzed the cytoskeleton of the cells after drug treatment. The control groups of both cell lines commonly appeared as fusiform cells, with microSelleckchem PD0332991 filaments that formed parallel stress fibers (Figures 5A-C, 6). After treatment with 0.4 mM cinnamic acid, the HT-144 cells showed a triangular or stellate morphology, and an altered orientation of actin filaments.

Future researches

should elucidate the specific context that is responsible for specific functions of miR-210. In addition, how to integrate multiple functionally Bindarit molecular weight different but related targets of one peculiar miRNA such as miR-210, so as to precisely predict its functions remains a great challenge. Besides functions of miR-210, we also reviewed the diagnostic and prognostic value of it. As described above, up-regulated miR-210 is not only be detected in cancer tissues, but also in body fluids. It is feasible to discriminate cancer from non-cancer with a specific group of miRNAs including miR-210. However, when it comes to prognosis, it is far Volasertib chemical structure too early to use miR-210 alone as a prognostic factor without dispute, and more investigations are needed to elucidate the underlying mechanism of such discrepancy. In future, global analysis of large cohorts of patients with not

only miRNAs expression profile but also mRNAs expression profile, even integrated with other genetic information such as DNA copy number variance, single nucleotide polymorphisms, will provide us more insights about significant prognostic find more factors as well as novel therapeutic targets. Acknowledgements This study was supported by National Natural Science Foundation of China (Grant no. 81272501). We acknowledge Dr. David L, Roerig for critical reading of the manuscript. References 1. Bartel DP: MicroRNAs: target recognition and regulatory functions. Cell 2009,136(2):215–233.PubMedCentralPubMed 2. Krol J, Loedige I, Filipowicz W: The widespread regulation of microRNA biogenesis, function and decay. Nat Rev

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