Abstract
Human embryonic carcinoma (EC; NCCIT) cells have self-renewal ability and pluripotency. Cancer stem cell markers are highly expressed in NCCIT cells, imparting them with the pluripotent nature to differentiate into other cancer types, including breast cancer. As one of the main cancer stem cell pathways, Wnt/β-catenin is also overexpressed in NCCIT cells. Thus, inhibition of these pathways defines the ability of a drug to target cancer stem cells. Tannic acid (TA) is a natural polyphenol present in foods, fruits, and vegetables that has anti-cancer activity. Through Western blotting and PCR, we demonstrate that TA inhibits cancer stem cell markers and the Wnt/β-catenin signaling pathway in NCCIT cells and through a fluorescence-activated cell sorting analysis we demonstrated that TA induces sub-G1 cell cycle arrest and apoptosis. The mechanism underlying this is the induction of mitochondrial reactive oxygen species (ROS) (mROS), which then induce the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated extrinsic apoptosis pathway instead of intrinsic mitochondrial apoptosis pathway. Moreover, ribonucleic acid sequencing data with TA in NCCIT cells show an elevation in TRAIL-induced extrinsic apoptosis, which we confirm by Western blotting and real-time PCR. The induction of human TRAIL also proves that TA can induce extrinsic apoptosis in NCCIT cells by regulating mROS.
Highlights
Human embryonic carcinoma (EC) cells exhibit similar gene expression profiles to those of embryonic stem (ES) cells, as both have an unlimited self-renewal ability and the capability to differentiate into all derivatives of the three embryonic germ layers—the ectoderm, endoderm, and mesoderm [1,2,3]
50 μM Tannic acid (TA) was used as the IC50 dosage for a time period of 48 h, which we used for further studies
To determine whether TA plays a role in the inhibition of stem cell markers, we checked the expression of stem cell markers SOX2, octamer-binding transcription factor 4 (OCT4), and NANOG in the mRNA level and obtained a significant concentration-dependent inhibition of these stem cell markers by TA in the NCCIT cells (Figure 1A,B)
Summary
Human embryonic carcinoma (EC) cells exhibit similar gene expression profiles to those of embryonic stem (ES) cells, as both have an unlimited self-renewal ability and the capability to differentiate into all derivatives of the three embryonic germ layers—the ectoderm, endoderm, and mesoderm [1,2,3]. As such, these cells can develop into several cell types in the adult human body [4]. The use of dietary phytochemicals originating from edible foods for cancer chemoprevention is a good method, as these natural substances lack side effects after treatment [14,15]
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