DataSheet_1_Integrated Whole Transcriptome Profiling and Bioinformatics Analysis for Revealing Regulatory Pathways Associated With Quercetin-Induced Apoptosis in HCT-116 Cells.pdf

Quercetin (QUE) is a bioactive component that belongs to the natural flavonoids group, and recent researchers found that it could prevent colorectal cancer (CRC). However, the exact mechanism by which QUE exerts its anti-tumor effects in CRC remains unclear. In this study, MTS assay and flow cytometry were used to detect the anti-tumor effects of QUE on HCT-116 cells. The results showed that QUE could inhibit the proliferation and induce apoptosis of HCT-116 cells. Furthermore, whole transcriptome sequencing was employed to establish the microRNA (miRNA), long non-coding RNA (lncRNA), circular RNA (circRNA), and mRNA profiles. A total of 240 differentially expressed lncRNAs (DElncRNAs), 131 circRNAs (DEcircRNAs), 83 miRNAs (DEmiRNAs), and 1415 mRNAs (DEmRNAs) were identified in the QUE-treated HCT-116 cells compared to the untreated HCT-116 cells. Then, quantitative real-time polymerase chain reaction (qRT-PCR) was used to validate the expression of selected circRNAs, miRNAs, lncRNAs, and mRNAs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed to further investigate RNAs’ biological functions and potential mechanisms. Based on the theory of competing endogenous RNA (ceRNA), the circRNA–miRNA–mRNA and lncRNA–miRNA–mRNA regulatory networks were constructed to illustrate the regulatory relationship between non-coding RNA (ncRNA) and mRNA. Our results provided novel information about the molecular basis of QUE in treating CRC. Our findings indicated that deep RNA sequencing analysis of mRNA and ncRNAs was a promising approach to research anticancer mechanisms.

槲皮素（Quercetin, QUE）是天然黄酮类化合物中的一种生物活性成分，近期研究发现其可预防结直肠癌（colorectal cancer, CRC）。然而，槲皮素在结直肠癌中发挥抗肿瘤作用的确切机制仍未阐明。本研究采用MTS实验与流式细胞术，检测槲皮素对HCT-116细胞的抗肿瘤作用。结果显示，槲皮素可抑制HCT-116细胞增殖并诱导其凋亡。进一步通过全转录组测序技术，构建了微小RNA（microRNA, miRNA）、长链非编码RNA（long non-coding RNA, lncRNA）、环状RNA（circular RNA, circRNA）以及信使RNA（messenger RNA, mRNA）的表达谱。与未经槲皮素处理的HCT-116细胞相比，经槲皮素处理的HCT-116细胞中共鉴定出240个差异表达长链非编码RNA（differentially expressed lncRNA, DElncRNA）、131个差异表达环状RNA（differentially expressed circRNA, DEcircRNA）、83个差异表达微小RNA（differentially expressed miRNA, DEmiRNA）以及1415个差异表达信使RNA（differentially expressed mRNA, DEmRNA）。随后采用实时定量聚合酶链反应（quantitative real-time polymerase chain reaction, qRT-PCR），对筛选得到的circRNA、miRNA、lncRNA及mRNA的表达水平进行验证。通过基因本体论（Gene Ontology, GO）与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）富集分析，进一步探究上述RNA的生物学功能与潜在作用机制。基于内源竞争RNA（competing endogenous RNA, ceRNA）理论，构建了circRNA–miRNA–mRNA与lncRNA–miRNA–mRNA调控网络，以阐明非编码RNA（non-coding RNA, ncRNA）与mRNA之间的调控关系。本研究结果为阐明槲皮素治疗结直肠癌的分子基础提供了全新视角。本研究表明，对mRNA与非编码RNA开展深度转录组测序分析，是研究抗肿瘤机制的一种极具潜力的方法。