Calycosin suppresses TGF-β-induced epithelial-to-mesenchymal transition and migration by upregulating BATF2 to target PAI-1 via the Wnt and PI3K/Akt signaling pathways in colorectal cancer cells

Abstract

Objectives To determine whether the upregulation of basic leucine zipper ATF-like transcription factor 2 (BATF2) by calycosin suppresses the growth and epithelial-to-mesenchymal transition (EMT) in human colorectal cancer (CRC) cells. Method Cells were cultured and treated with different concentrations of calycosin for different periods of time. Protein and mRNA expression was determined by western blotting and quantitative PCR. Cell migration was assessed by Transwell experiments. Co-immunoprecipitation and luciferase assays were used to analyze the association between BATF2 and plasminogen activator inhibitor-1. (PAI-1). Cell apoptosis was determined by flow cytometry; β-catenin cellular localization was visualized by immunofluorescent staining. Results Calycosin up-regulated the expression of BATF2 via the signal transducer and activator of transcription 3 (STAT3) pathway, which was antagonized by transforming growth factor beta (TGF-β), calycosin promoted the cell apoptosis and growth inhibition via phosphoinositide 3-kinase (PI3K)/Akt pathway. TGF-β promoted cell growth, which was inhibited by calycosin regulating the expression of proliferating cell nuclear antigen (PCNA) via the phosphoinositide 3-kinase pathway. TGF-β suppressed expression of BAX via the phosphoinositide 3-kinase pathway but induced cell apoptosis .calycosin enhanced the effect of TGF-β on cell apoptosis,In addition, calycosin suppressed TGF-β-induced cell migration by increasing BATF2 to target PAI-1. TGF-β-induced EMT was inhibited by calycosin in human CRC LoVo and HCT116 cell lines via the Wnt signaling pathway. Conclusions The induction of BATF2 by calycosin may be a feasible therapeutic option for CRC. Graphical Abstract .

Emerging Roles of Wnt Ligands in Human Colorectal Cancer

“…Interestingly, the effect of EGCG on CRC could be attributed to the inhibition on CRC stem cells by suppressing the canonical Wnt signaling ( 155 ). Furthermore, other natural compounds such as calycosin, isobavachalcone, resveratrol, etc., were also proved to suppress the malignant phenotypes of CRC via inhibiting the Wnt signaling pathways ( 156 – 158 ), providing more feasible therapeutic options for CRC treatment. However, few natural compounds have been proven to suppress the development of CRC by directly targeting Wnts, and none of them is currently undergoing clinical testing.…”

CX3CR1 regulates hepatocellular carcinoma(HCC) metastasis via PI3K/AKT pathway

“…Chemokines and their receptors have been proved to play a very important role in tumor microenvironment. Studies have confirmed that chemokine axis CXCL9/10-CXCR3, CXCL12-CXCR4, CCL20-CCR6, CCL5-CCR5, CXCL5-CXCR2 and so on can regulate the functions of various immune cells in tumor microenvironment to further promote immune escape of tumor cells and angiogenesis of tumors to help metastasis and progress of tumors [35][36][37][38][39]. CX 3 CR1 has also been shown to play an important role in regulating tumor and tumor microenvironment [40].…”

<p>Salinomycin and Sulforaphane Exerted Synergistic Antiproliferative and Proapoptotic Effects on Colorectal Cancer Cells by Inhibiting the PI3K/Akt Signaling Pathway in vitro and in vivo</p>

“…For the migration assay, 10×10 4 cells in 200 μL of FBS-free medium were added to the upper chamber, which were not surface-coated with Matrigel matrix; all other aspects of the procedures were the same. 24…”

“…Quantitative RT-PCR (10-μL reaction volume) was performed using SYBR Premix Ex Taq (TaKaRa, Shiga, Japan). 24 β-Actin expression was used as an internal control. The gene-specific primers (BioSune, Shanghai, China) used in this study are listed in Table 1.…”