(E)-4-(3-(3,5-dimethoxyphenyl)allyl)-2-methoxyphenol inhibits growth of colon tumors in mice

Abstract

In our previous study, we found that (E)-2,4-bis(p-hydroxyphenyl)-2-butenal showed anti-cancer effect, but it showed lack of stability and drug likeness. We have prepared several (E)-2,4-bis(p-hydroxyphenyl)-2-butenal analogues by Heck reaction. We selected two compounds which showed significant inhibitory effect of colon cancer cell growth. Thus, we evaluated the anti-cancer effects and possible mechanisms of one compound (E)-4-(3-(3,5-dimethoxyphenyl)allyl)-2-methoxyphenol in vitro and in vivo. In this study, we found that (E)-4-(3-(3,5-dimethoxyphenyl)allyl)-2-methoxyphenol induced apoptotic cell death in a dose dependent manner (0-15 μg/ml) through activation of Fas and death receptor (DR) 3 in HCT116 and SW480 colon cancer cell lines. Moreover, the combination treatment with (E)-4-(3-(3,5-dimethoxyphenyl)allyl)-2-methoxyphenol and nuclear factor κB (NF-κB) inhibitor, phenylarsine oxide (0.1 μM) or signal transducer and activator of transcription 3 (STAT3) inhibitor, Stattic (50 μM) increased the expression of Fas and DR3 more significantly. In addition, (E)-4-(3-(3,5-dimethoxyphenyl)allyl)-2-methoxyphenol suppressed the DNA binding activity of both STAT3 and NF-κB. Knock down of STAT3 or NF-κB p50 subunit by STAT3 small interfering RNA (siRNA) or p50 siRNA magnified (E)-4-(3-(3,5-dimethoxyphenyl)allyl)-2-methoxyphenol-induced inhibitory effect on colon cancer cell growth. Besides, the expression of Fas and DR3 was increased in STAT3 siRNA or p50 siRNA transfected cells. Moreover, docking model and pull-down assay showed that (E)-4-(3-(3,5-dimethoxyphenyl)allyl)-2-methoxyphenol directly bound to STAT3 and NF-κB p50 subunit. Furthermore, (E)-4-(3-(3,5-dimethoxyphenyl)allyl)-2-methoxyphenol inhibited colon tumor growth in a dose dependent manner (2.5 mg/kg-5 mg/kg) in mice. Therefore, these findings indicated that (E)-4-(3-(3,5-dimethoxyphenyl)allyl)-2-methoxyphenol may be a promising anti-cancer agent for colon cancer with more advanced research.

Piperlongumine inhibits lung tumor growth via inhibition of nuclear factor kappa B signaling pathway

“…Similar mechanism was found in the inhibitory effect on cancer cell growth of thiacremonone and tectochrysin that bound to the DNA binding site of p50 to inhibit the DNA binding activity of NF-κB in NSCLC cells 26 27 . Besides, (E)-2,4-Bis ( p -hydroxyphenyl)-2-butenal inhibits NF-κB activity via directly binding to IKKβ 28 , (E)-4-(3-(3,5-dimethoxyphenyl)allyl)-2-methoxyphenol inhibits NF-κB activity via directly binding to DNA binding site of p50 29 . Melittin, the major component of bee venom directly bound to the nuclear localization sequence (NLS) which was near to the C-terminus of p50 to block NF-κB activation 30 .…”

A C21-steroidal derivative suppresses T-cell lymphoma in mice by inhibiting SIRT3 via SAP18-SIN3

“…To further confirm the binding of A671 to SAP18, pool-down experiments performed using epoxy-activated Sepharose 6B (ES6B) beads 35 , 36 . Extracts isolated from HEL cells incubated with conjugated A671 + ES6B or ES6B beads alone, and after washing, the eluted proteins subjected to western blotting.…”

A small molecule, (E)-2-methoxy-4-(3-(4-methoxyphenyl) prop-1-en-1-yl) phenol suppresses tumor growth via inhibition of IkappaB kinase β in colorectal cancer in vivo and in vitro

“…HCT116 and SW480 colon cancer cells were treated with MMPP (0-15 μg/mL) for 24 h, then were homogenized with a protein extraction solution (PRO-PREP™, iNtRON Biotechnology, Seongnam, Gyeonggi, Korea), and lysed by 60 min incubation on ice. Western blotting was described elsewhere [ 36 ]. After a short washing in TBST, the membranes were immunoblotted with the following primary antibodies: caspase-3 and caspase-8 (1:1000 dilutions; Cell Signaling, Beverly, MA, USA) and Fas, DR3, DR4, DR5, DR6, IKKβ, p-IKKβ, IκBα, p-IκBα, p50, p65, Bcl-2, Bax, Histone-H1 and β-actin (1:1000 dilutions; Santa Cruz Biotechnology, Santa Cruz, CA, USA).…”

“…Animals were sacrificed by CO 2 , tumor tissues were collected. Immunohistochemistry assay was performed as described elsewhere [ 36 ]. Staining was carried out using antibodies against PCNA, p-IKKβ, active caspase-3, DR5 and DR6 (1:500, Abcam, Cambridge, UK).…”

“…MMPP was conjugated with epoxy-activated Sepharose 6B. Pull-down assay was performed as described elsewhere [ 36 ]. MMPP (1 mg) was dissolved in 1mL of coupling buffer (0.1 M NaHCO 3, pH 13.0 containing 0.5 M NaCl).…”