Withaferin A targeting both cancer stem cells and metastatic cancer stem cells in the UP-LN1 carcinoma cell model

Abstract

As our understanding of cancer stem cell (CSC) biology improves, search for inhibitory agents of CSCs and metastatic CSCs (mCSCs) positive for CXCR4 is warranted. Withaferin A (WA), a withanolide extracted from the medicinal plant Withania somnifera, has been shown to exhibit anti-cancer effects through multiple mechanisms. Whether WA could selectively target CSCs, mCSCs, or non-CSCs of a gastrointestinal (GI) carcinoma tumor remains unclear. Methods: Side-population (SP) analysis, flow cytometric phenotyping and sorting, non-invasive imaging in conjunction with xenotransplantation, and immunohistology were used in this investigation. Results: Using the lymph node metastatic GI cancer cell line UP-LN1, consisting of CD44 high /CD24 low floating (F) and CD44 low /CD24 high adherent (A) cell subsets, this study demonstrated that as compared with parental UP-LN1 cells or A cells, WA preferentially reduced F-cell proliferation, tumor sphere formation, and SP cells in vitro in greater effi ciencies by apoptosis. This action was mechanistically mediated via the down-regulation of CXCR4/CXCL12 and STAT3/interleukin-6 axes, both of which are instrumental in the acquisition of metastatic ability. Attenuation of interferon-γ-induced CXCR4 expression in F cells by knockdown with siRNA or blocking with an anti-CXCR4 antibody, followed by Western blot analysis, showed signifi cantly reduced metastatic potential in vitro. The extent of in vitro anti-invasive effect of WA on the IFN-γ-treated F cells was signifi cantly greater than on the F cells without WA treatment, or F cells treated with control siRNA or with control IgG antibody. The observed in vitro effects of WA on the CSC and mCSC targeting were validated by data obtained with non-invasive imaging in NOD/SCID mouse xenotransplantation. Conclusion: WA could effi ciently block the formation of both CSCs and mCSCs in the UP-LN1 cell line, suggesting that WA may be considered an effective therapeutic agent for this type of GI malignancies.

Cancer stem cell markers predict a poor prognosis in renal cell carcinoma: a meta-analysis

“…Accumulated evidence shows that cancer can be considered as a stem cell disease [ 44 ]. CSCs, which comprise a small subpopulation of cancer cells, exhibit self-renewal ability and cancer-propagating capacity [ 45 , 46 ]. The concept of contribution of CSCs to cancer initiation and therapeutic resistance is widely accepted, so a better understanding of the characteristics of CSCs will provide valuable therapeutic and prognostic targets for clinical practice.…”

The Potential Pharmacological Effects of Natural Product Withaferin A in Cancer: Opportunities and Challenges for Clinical Translation

“…WA inhibited the sphere formation and induced apoptosis of the F cell population in the UP-LN1 lymph node metastatic cell line (gastrointestinal carcinoma cell line). One more reason for hiding the apoptosis by WA is the lower levels of glutathione in the F cell sub-population [72].…”

Withaferin A Increases the Effectiveness of Immune Checkpoint Blocker for the Treatment of Non-Small Cell Lung Cancer

“…Withaferin A (WFA) is a broad-spectrum anticancer steroidal lactone isolated from the leaves and roots of Withania Somnifera (WS) [ 22 ]. Mechanistically, WFA induces apoptotic and ferroptotic cell death, and inhibits angiogenesis, metastasis, and cancer stemness [ 23 , 24 , 25 ]. Additionally, WFA exerts its anti-cancer effects by targeting several downstream molecules.…”