buy SGX523 Our previous study demonstrated that HDAC

Our previous study demonstrated that HDAC inhibition contributed to the anticancer effect of statins (Lin et al., 2008). Combining anti-cancer drugs with different mechanisms is a rational approach to improve efficacy. JMF3086, synthesized to inhibit both HDACs and HMGR, exhibits significant benefits above lovastatin plus SAHA. This advantage is like a two-in-one antibody which is better than two monospecific buy SGX523 (Schaefer et al., 2011). Various polypharmacological molecules dually inhibiting HDACs and other therapeutic targets have also been developed and provide an impetus to develop more anti-cancer treatments (Falkenberg and Johnstone, 2014). Evading apoptosis is a hallmark of cancer (Hanahan and Weinberg, 2011), where apoptosis induction represents one therapeutic strategy (Cotter, 2009). Anti-apoptotic genes such as BCL2 and MCL1 were down-regulated by JMF3086 from genome-wide ChIP-on-chip and Q-PCR analyses, and JMF3086 did induce apoptosis in HCT116 cells and tumors of CRC mice, indicating the contribution of apoptosis induction to the therapeutic effect of JMF3086 on CRC. NR3C1, which is a glucocorticoid receptor, was predicted to be induced by JMF3086 from GO and IPA analyses. It induced apoptotic cell death via decreasing the expression of anti-apoptotic proteins such as BCL2 and MCL1, and/or via inducing the expression of pro-apoptotic proteins such as BCL-2-like apoptosis initiator-11 (BCL2L11) (Schlossmacher et al., 2011). JMF compounds did induce NR3C1 mRNA expression in CRC mice, indicating its contribution to apoptosis. The tumor microenvironment consisting of cancer cells, stromal tissue, blood vessels, immune cells, fibroblasts, bone marrow-derived inflammatory cells, lymphocytes and the extracellular matrix, plays a critical role in cancer development, progression and metastasis (Popivanova et al., 2009; Hanahan and Weinberg, 2011). Clinical studies have indicated its association with poor prognosis in patients with various human cancers (Erreni et al., 2011; Gregory and Houghton, 2011). Tumor-associated macrophages (TAMs) are key components of tumor microenvironment (Noy and Pollard, 2014). They can produce growth factors, cytokines, chemokines and matrix metalloproteases (MMP) to promote cancer progression, metastasis, angiogenesis and lymphangiogenesis (Qian and Pollard, 2010). Tumor-associated neutrophils (TANs) acting through the production of cytokines/chemokines and the recruitment of TAMs to the tumor sites also play a role in tumor microenvironment (Mantovani, 2009; Nathan, 2006). In this study, both infiltrations of macrophages and neutrophils occurring in AOM-DSS mouse models were blocked by JMF compounds but not by statins, SAHA or lovastatin plus SAHA (Fig. 3E and F), probably explaining the better efficacy of JMF3086 than lovastatin plus SAHA in treating AOM-DSS CRC. However, whether TAMs and TANs affected by JMF3086 are beyond its inhibitions on HMGR and HDAC remains to be investigated. Metastasis is the major cause of CRC-related deaths, and livers or lungs are the primary organs of metastasis (Villeneuve and Sundaresan, 2009; Edwards et al., 2012). JMF3086 showed significant efficacy in inhibiting metastasis and potentiated the effect of oxaliplatin. The mechanism may involve the inhibition on angiogenesis (Fig. 4G). Furthermore, combination of JMF3086 with oxaliplatin reduced the hypoxic region and downregulated HIF-1α expression in vivo (Fig. 6I). Stemness of cancer cells may play a role in the genesis, maintenance, recurrence, metastasis, and drug resistance of CRC (Eyler and Rich, 2008; Pignalosa and Durante, 2012). Loss of CSC populations or stem-like properties could hinder the metastatic spread of the tumor (Pattabiraman and Weinberg, 2014). Stem cell expansion was derived from chronic inflammation causing damage and regeneration of crypt and release of cytokines, which maintain and regulate cancer stem-cell niche (Carpentino et al., 2009; Liu and Wicha, 2010). JMF3086 attenuated the cytokines in CRC mice and inhibited the expression of stemness-related genes. We provide direct evidences to demonstrate that HMGR enhances the stemness of CRC cells, and JMF3086 inhibited the growth of colonospheres and colonosphere-derived xenografts more than lovastatin plus SAHA. Statins was reported to lower cytokines levels to target CSCs and inhibited tumor growth as well as metastasis (Ridker et al., 2005), and HDAC inhibitor reduced the stemness of CRC cells (Kodach et al., 2011; Sikandar et al., 2010). These indicate that dual HMGR-HDAC inhibitor JMF3086-induced inhibitions on stemness and metastasis contribute to its anti-CRC effect.