RESUMO
To understand how upregulated isoglutaminyl cyclase (isoQC) is involved in the initiation of diseases such as cancer, we developed a human KYSE30 carcinoma cell model in which isoQC was stably overexpressed. GO and KEGG analysis of the DEGs (228) and DEPs (254) respectively implicated isoQC on the proliferation invasion and metastasis of cells and suggested that isoQC might participate in the regulation of MAPK, RAS, circadian rhythm, and related pathways. At the functional level, isoQC-overexpressing KYSE30 cells showed enhanced proliferation, migration, and invasion capacity. Next, we decided to study the precise effect of isoQC overexpression on JNK, p-JNK, AKT, p-AKT, ERK, p-ERK, and PER2, as RNA levels of these proteins are significantly correlated with signal levels indicated in RNA-Seq analysis, and these candidates are the top correlated DEPs enriched in RT-qPCR analysis. We saw that only p-ERK expression was inhibited, while PER2 was increased. These phenotypes were inhibited upon exposure to PER2 inhibitor KL044, which allowed for the restoration of p-ERK levels. These data support upregulated isoQC being able to promote cancer cell proliferation and migration in vitro, likely by helping to regulate the MAPK and RAS signaling pathways, and the circadian protein PER2 might be a potential mediator.
Assuntos
Aminoaciltransferases , Movimento Celular , Proliferação de Células , Sistema de Sinalização das MAP Quinases , Humanos , Proliferação de Células/genética , Movimento Celular/genética , Sistema de Sinalização das MAP Quinases/genética , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Proteínas Circadianas Period/genética , Proteínas Circadianas Period/metabolismo , Invasividade Neoplásica , Regulação para Cima , Neoplasias/genética , Neoplasias/patologia , Neoplasias/metabolismoRESUMO
The dried roots and rhizomes of Radix Gentianae Macrophyllae are widely used as food material or medicinal crops. "Sweating" is a traditional postharvest processing method, the basic processing procedure consists of softening, stacking and drying. The aim of this paper is to unveil the scientific connotation responsible for the "Sweating" processing in Radix Gentianae Macrophyllae during postharvest. Thus, the effect of different postharvest processing methods on the metabolic pathways of Radix Gentiasnae Macrophyllae was studied by the non-targeted metabolomic technique in combination with the label-free proteomics approach. The results showed that the differentially accumulated metabolites and abundant proteins were mainly enriched in the pathways of phenylalanine, tyrosine and tryptophan biosynthesis, polyphenols and terpenoids biosynthesis. "Sweating" has a greater up-regulation effect on these pathways than "Non-sweating", and can induce protein expression and metabolite accumulation associated with the quality traits of Radix Gentianae Macrophyllae. The results provide a detailed explanation of the scientific connotation of crucial steps of "Sweating" processing wherein opportunities existed for taking appropriate measures to enhance the accumulation of bioactive ingredients. These findings will serve as significant references for enhancing the postharvest processing technology of Radix Gentianae Macrophyllae and similar plants, resulting in higher product quality for food or plant materials production.