Evodiamine Eliminates Colon Cancer Stem Cells via Suppressing Notch and Wnt Signaling

Abstract

Evodiamine, an alkaloid contained in traditional Asian herbal medicines that have been used for hundreds years, is interesting due to its cytotoxic effects against many cancers. We examined the effect of evodiamine on the cancer stem cell (CSC) population and the bulk cultured cancer cells (BCC) of colon cancers to examine the double targeting effect. We found that three colon cancer cell lines’ BCC and CSC are effectively targeted by evodiamine. Evodiamine was able to suppress BCC proliferation and induce apoptosis of the cells captured in G2/M phase, as previously reported. However, evodiamine did not cause the accumulation of CSCs at a certain stage of the cell cycle, resulting in the elimination of stemness through an unknown mechanism. By analyzing the expression of 84 genes related to CSCs in two colon cancer cell lines’ CSC, as well as performing further informatics analyses, and quantitative RT-PCR analyses of 24 CSC genes, we found that evodiamine suppressed the expression of the genes that control key signaling pathways of CSC, namely, WNT and NOTCH signaling, to lead CSC elimination. These results suggest that evodiamine should be further developed for targeting both BCCs and CSCs in colon cancers.

Targeting a Lipid Desaturation Enzyme, SCD1, Selectively Eliminates Colon Cancer Stem Cells through the Suppression of Wnt and NOTCH Signaling

“…They were maintained as bulk culture cells (BCC) in Dulbecco’s modified eagle’s medium (DMEM, Invitrogen, Waltham, MA, USA) or RPMI 1640 (Invitrogen) supplemented with FBS (Fetal bovine serum, Invitrogen). For cancer stem cell (CSC) enrichment, they were cultured in DMEM/F12 containing B27 supplement and 20 ng/mL EGF (all from Invitrogen) in a polyHEMA (Sigma–Aldrich, St. Louis, MO, USA)-coated dish as described previously [ 20 ]. All cultured cells were maintained at 37 °C in a humidified atmosphere of 5% CO 2 .…”

“…The cells were reverse transfected with siSCD1, and siControl (NC) obtained from IDT (Coralville, IA, USA) using Lipofectamine ® RNAiMAX (Thermo, Waltham, MA, USA) in accordance with the manufacturer’s protocol. Cell viability was evaluated as previously published [ 20 ]. Four thousand cells were seeded in every well of a 96-well plate.…”

“…The cell lysates were prepared as described previously [ 20 ]. Briefly, the Tris-based buffer containing phosphatase inhibitors and protease inhibitors (Roche, Basel, Switzerland) was applied and cleared with centrifugation.…”

Emerging agents that target signaling pathways in cancer stem cells

“…Verteporfin can inhibit the transcriptional activity of YAP/TAZ-TEAD, reduce the expression of CSC markers, and suppress CSC proliferation [108,109]. Evodiamine (Evo), which is isolated from the Chinese herb Evodia rutaecarpa Benham, can activate MST1/2-mediated phosphorylation of LATS1/2, which leads to YAP/TAZ phosphorylation and prevents YAP/TAZ translocation from the cytoplasm into the nucleus, and it has been shown that Evo can inhibit the proliferation of colon CSCs [110,111]. Additionally, tanshinone IIA and limonin, which are extracted from Chinese herbs, have been shown to attenuate the stemness of cervical carcinoma stem cells by inhibiting the cytoplasmic-nuclear translocation of YAP [112,113].…”

PTEN, a Barrier for Proliferation and Metastasis of Gastric Cancer Cells: From Molecular Pathways to Targeting and Regulation

“…As a potential factor in cell fate, increasing evidence has shown that Notch signaling plays like a double-edged sword in cancer, and it has both oncogene and onco-suppressor activities [ 248 , 249 , 250 , 251 , 252 , 253 , 254 ]. It has been reported that metastasis of GC cells is tightly regulated by Notch/PTEN/Akt axis.…”