Identification of Two Novel CIL-102 Upregulations of ERP29 and FUMH to Inhibit the Migration and Invasiveness of Colorectal Cancer Cells by Using the Proteomic Approach

Abstract

CIL-102 (1-[4-(furo[2,3-b]quinolin-4-ylamino) phenyl]ethanone) is a major active agent of Camptotheca acuminata’s alkaloid derivative, and its anti-tumorigenic activity, a valuable biological property of the agent, has been reported in many types of cancer. In this study, we researched the novel CIL-102-induced protein for either the induction of cell apoptosis or the inhibition of cell migration/invasiveness in colorectal cancer cells (CRC) and their molecular mechanism. Firstly, our data showed that CIL-102 treatment not only increased the cytotoxicity of cells and the production of Reactive Oxygen Species (ROS), but it also decreased cell migration and invasiveness in DLD-1 cells. In addition, many cellular death-related proteins (cleavage caspase 9, cleavage caspase 3, Bcl-2, and TNFR1 and TRAIL) and JNK MAPK/p300 pathways were increased in a time-dependent manner. Using the proteomic approach with a MALDI-TOF-TOF analysis, CIL-102-regulated differentially expressed proteins were identified, including eight downregulated and 11 upregulated proteins. Among them, upregulated Endoplasmic Reticulum resident Protein 29 (ERP29) and Fumarate Hydratase (FUMH) by CIL-102 were blocked by the inhibition of ROS production, JNK activity, and p300/CBP (CREB binding protein) signaling pathways. Importantly, the knockdown of ERP29 and FUMH expression by shRNA abolished the inhibition of cell migration and invasion by CIL-102 in DLD-1 cells. Together, our findings demonstrate that ERP29 and FUMH were upregulated by CIL102 via ROS production, JNK activity, and p300/CBP pathways, and that they were involved in the inhibition of the aggressive status of colorectal cancer cells.

Euchromatic histone lysine methyltransferase 2 facilitates radioresistance in prostate cancer by repressing endoplasmic reticulum protein 29 transcription

“…Upregulation of ERP29 in breast cancer cells inhibits cell proliferation and invasion by upregulating E‐cadherin, cyclin‐dependent kinase inhibitor, and spleen tyrosine kinase; downregulating fibronectin, epidermal growth factor receptor, and plasminogen activator receptor; and attenuating the overall ERK cascade 32 . A previous study suggested that the inhibitory effects of CIL‐102, an alkaloid derivative, on the migration and invasion of colorectal cancer cells are abrogated by ERP29 knockdown through ROS, JNK, and p300/CBP pathways 14 . Overexpression of ERP29 prominently inhibits the proliferative capacity of gastric cancer cells and nullifies the epithelial‐mesenchymal transition process in gastric cancer by inhibiting ERK1/2 and AKT phosphorylation 12 .…”

“…A previous study revealed that ERP29 knockdown elevates the migratory and invasive abilities of gastric cancer cells 13 . It has been documented that ERP29 upregulation inhibits migration and invasion of colon cancer cells 14 . Moreover, ERP29 protein expression in androgen‐sensitive prostate cancer cells (LNCaP) is increased three‐fold by co‐treatment with dihydrotestosterone and EPI‐001 (a prostate cancer inhibitor), 15 suggesting that ERP29 may be an antitumor protein in prostate cancer.…”

“…13 It has been documented that ERP29 upregulation inhibits migration and invasion of colon cancer cells. 14 Moreover, ERP29 protein expression in androgen-sensitive prostate cancer cells (LNCaP) is increased threefold by co-treatment with dihydrotestosterone and EPI-001 (a prostate cancer inhibitor), 15 suggesting that ERP29 may be an antitumor protein in prostate cancer. During liver regeneration, ERP29 mRNA is modified by m 6 A, 16 but histone methylation of the ERP29 gene remains an unexplored area.…”

Antrodin C Isolated from Antrodia Cinnamomea Induced Apoptosis through ROS/AKT/ERK/P38 Signaling Pathway and Epigenetic Histone Acetylation of TNFα in Colorectal Cancer Cells

“…3-(4,5-Di-2-yl)-2,5-ditetrazoliu (MTT) was used for cell viability analysis to evaluate the cytotoxic effect, as previously described [ 24 ]. To observe cell morphology characteristic of apoptosis, 4,6-diamidino-2-phenylindole (DAPI)-stained cells were examined by fluorescence microscopy, as previously described [ 25 ]. Cell-cycle distribution was analyzed using flow cytometry, as previously described [ 26 ].…”

2D-DIGE-MS Proteomics Approaches for Identification of Gelsolin and Peroxiredoxin 4 with Lymph Node Metastasis in Colorectal Cancer

“…MS spectra and MS/MS fragment ion mass were determined with a Bruker MALDI-TOF/TOF analyzer (Bruker Daltonics, Bremen, Germany), and all product ions were submitted to a computer database and analyzed with the MASCOT MS/MS ion search (Matrix Science Inc., MA, USA) using the SwissProt database (all entries). The Bruker operation system, namely flexControl Version 3.3 (Build 85) and flexAnalysis software, were used to create peak lists [21].…”