Artesunate inhibits intestinal tumorigenesis through inhibiting wnt signaling

Abstract

Artesunate (ART) is a clinically approved antimalarial drug and was revealed as a candidate of colorectal cancer chemopreventive agents in our drug screening system. Here, we aimed to understand the suppressive effects of ART on intestinal tumorigenesis. In vitro, ART reduced T-cell factor/lymphoid enhancer factor (TCF/LEF) promoter transcriptional activity. In vivo, ART inhibited intestinal polyp development. We found that ART reduces TCF1/TCF7 nuclear translocation by binding the Ras-related nuclear protein (RAN), suggesting that ART inhibits TCF/LEF transcriptional factor nuclear translocation by binding to RAN, thereby inhibiting Wnt signaling. Our results provide a novel mechanism through which artesunate inhibits intestinal tumorigenesis.

Theoretical Basis Validation and Oxidative Stress Markers for Cancer Prevention Clinical Trials of Aspirin

“…Both c-Myc and Cyclin D1 are cell proliferation-related factors that are induced by activation of Wnt signaling through APC gene mutations found in ~ 80% of CRC (21,22). So far, it has been suggested that suppression of excessive Wnt signaling is involved in polyp inhibition experiments using Min mice (23)(24)(25). Previous studies reported that aspirin suppressed the expression levels of c-Myc and Cyclin D1 in many cancer cells and also suppressed cell proliferationrelated marker proteins, such as Ki67 and PCNA (26-28).…”

“…The other regions were opened longitudinally and xed at between sheets of lter paper in 10% buffered formalin. Polyp number, size, and intestinal distributions were assessed using a stereoscopic microscope (23). All experiments were performed in accordance with the "Guidelines for Animal Experiments in the National Cancer Center" and were approved by the Institutional Ethics Review Committee for Animal Experimentation of the National Cancer Center.…”

“…Cells stably expressing TCF/LEF-, NF-κB-, and NRF2-Luc were treated with hygromycin and cloned. These cells are referred to as HCT116-TCF/LEF-Luc, HCT116-NF-κB-Luc, and HCT116-NRF2-Luc cells, respectively (23). For the aspirin experiments, the HCT116-TCF/LEF-Luc, HCT116-NF-κB-Luc, and HCT116-NRF2-Luc cells were seeded in 96-well plates (2 × 10 4 cells per well).…”

“…Total RNA was isolated using RNAiso Plus (Takara Bio, Shiga, Japan) and 1-µg aliquots in a nal volume of 20 µL were used for cDNA synthesis using a High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA). Real-time PCR was carried out using the CFX96/384 PCR Detection System (Bio-Rad, Tokyo, Japan) and FastStart Universal SYBR Green Mix (Roche Diagnostics, Mannheim, Germany), in accordance with the manufacturers' instructions (23). The designed gene primer sequences used to evaluate human and mouse mRNA levels are shown in Supplementary Table 1.…”

Theoretical basis validation and oxidative stress markers for cancer prevention clinical trials of aspirin

“…Both c-Myc and Cyclin D1 are cell proliferation-related factors that are induced by activation of Wnt signaling through APC gene mutations found in ~ 80% of CRC 21 , 22 . So far, it has been suggested that suppression of excessive Wnt signaling is involved in polyp inhibition experiments using Min mice 23 – 25 . Previous studies reported that aspirin suppressed the expression levels of c-Myc and Cyclin D1 in many cancer cells and also suppressed cell proliferation-related marker proteins, such as Ki67 and PCNA 26 – 28 .…”