Apigenin Inhibits the Growth of Hepatocellular Carcinoma Cells by Affecting the Expression of microRNA Transcriptome

Abstract

BackgroundApigenin, as a natural flavonoid, has low intrinsic toxicity and has potential pharmacological effects against hepatocellular carcinoma (HCC). However, the molecular mechanisms involving microRNAs (miRNAs) and their target genes regulated by apigenin in the treatment of HCC have not been addressed.ObjectiveIn this study, the molecular mechanisms of apigenin involved in the prevention and treatment of HCC were explored in vivo and in vitro using miRNA transcriptomic sequencing to determine the basis for the clinical applications of apigenin in the treatment of HCC.MethodsThe effects of apigenin on the proliferation, cell cycle progression, apoptosis, and invasion of human hepatoma cell line Huh7 and Hep3B were studied in vitro, and the effects on the tumorigenicity of Huh7 cells were assessed in vivo. Then, a differential expression analysis of miRNAs regulated by apigenin in Huh7 cells was performed using next-generation RNA sequencing and further validated by qRT-PCR. The potential genes targeted by the differentially expressed miRNAs were identified using a curated miRTarBase miRNA database and their molecular functions were predicted using Gene Ontology and KEGG signaling pathway analysis.ResultsCompared with the control treatment group, apigenin significantly inhibited Huh7 cell proliferation, cell cycle, colony formation, and cell invasion in a concentration-dependent manner. Moreover, apigenin reduced tumor growth, promoted tumor cell necrosis, reduced the expression of Ki67, and increased the expression of Bax and Bcl-2 in the xenograft tumors of Huh7 cells. Bioinformatics analysis of the miRNA transcriptome showed that hsa-miR-24, hsa-miR-6769b-3p, hsa-miR-6836-3p, hsa-miR-199a-3p, hsa-miR-663a, hsa-miR-4739, hsa-miR-6892-3p, hsa-miR-7107-5p, hsa-miR-1273g-3p, hsa-miR-1343, and hsa-miR-6089 were the most significantly up-regulated miRNAs, and their key gene targets were MAPK1, PIK3CD, HRAS, CCND1, CDKN1A, E2F2, etc. The core regulatory pathways of the up-regulated miRNAs were associated with the hepatocellular carcinoma pathway. The down-regulated miRNAs were hsa-miR-181a-5p and hsa-miR-148a-3p, and the key target genes were MAPK1, HRAS, STAT3, FOS, BCL2, SMAD2, PPP3CA, IFNG, MET, and VAV2, with the core regulatory pathways identified as proteoglycans in cancer pathway.ConclusionApigenin can inhibit the growth of HCC cells, which may be mediated by up-regulation or down-regulation of miRNA molecules and their related target genes.

The Effect of Encapsulated Apigenin Nanoparticles on HePG-2 Cells through Regulation of P53

“…Several studies established the enhanced cytotoxic activity of Ap against cancer cells after being loaded on different nano-carrier systems [21,25,49,58,97]. In our study, we loaded the Ap onto ChNPs, then coated them with albumin-folic to produce water-soluble nanoparticles and targeted the cancer cells by increasing the penetration of Ap inside the HePG-2 cell.…”

Influence of the Bioactive Diet Components on the Gene Expression Regulation

“…and hsa-miR-148a-3p apigenin hepatocellular carcinoma [128] ↑ miR-34a-5p lung cancer [129] ↑ miR-152-5p cervical carcinoma [130] ↑ miRNA-215-5p colorectal cancer [131] ↑ miR-34a-5p luteolin lung cancer [132] ↑ miR-203 breast cancer [133] ↑ miR-6809-5p hepatocellular carcinoma [134] ↑ miRNA-34a gastric cancer [135] ↓ miR-21 and ↑ miR-16 and -34a breast cancer [136] ↓ microRNA-132 Bronchopneumonia [137] ↓ miRNA-301-3p pancreatic cancer [138] ↑ microRNA-340 kaempferol lung cancer [139] ↑ miR-339-5p colon cancer [140] ↓ microRNA-21 liver cancer [141] ↓ miR-146a osteoarthritis [142] ↑ miR-203 hypertension [143] ↑ miR-132 and miR-502c chrysin breast cancer [144] ↑ miR-9 and Let-7 gastric cancer [145] ↓ miR-18a, miR-21, and miR-221 genes gastric cancer [146] ↓ microRNA (miR)-92a atherosclerosis [147] ↓ miR-636 caffeic acid diabetic nephropathy [148] ↑ miR-221 epigallocatechin-3gallate hepatic fibrosis [149] ↑ miR-548m hepatitis C [150] ↑ microRNA-let-7b melanoma [151] ↑ miR-520a-3p prostate cancer [152] ↑ miR-384 ischemic heart disease [153] ↓ miR-25 breast cancer [154] ↑ miR-9-3 sulforaphane lung cancer [155] ↑ miR135b-5p pancreatic cancer [156] ↓ miRNA-423-5p liver fibrosis [157] ↓ miR30a-3p pancreatic cancer [158] ↓ miR-155 acute myeloid leukemia [159] ↓ miR-21 colon cancer ↑ miRNA-124-3p nasopharyngeal cancer [160] ↓ miR-23b, miR-92b, miR-381, and miR-382 breast cancer [36] ↑ miR-29a-3p and miR-200a Carotenoids: lycopene, β-carotene, lutein, astaxanthin colorectal cancer [161] ↑ miR-320d, miR-1246 and miRNA-1...…”

Immune-related lincRNA pairs predict prognosis and therapeutic response in hepatocellular carcinoma

“…Apigenin, a natural flavonoid, has low intrinsic toxicity and possesses preventive and therapeutic potential against cancers 50 . In HCC, apigenin could inhibit the growth and epithelial-mesenchymal transition of cancer cells 51 , 52 .…”