Transcriptomic Analysis on the Key Genes and Functional Pathways of Ramulus Cinnamomi in Repressing the Proliferation of Lung Cancer in vitro

Abstract

Background: This study is engineered to investigate the key genes and functional pathways of Ramulus Cinnamomi (RC) in inhibiting the proliferation of lung cancer cell lines A549 and H1650 in vitro. Methods The total RNA in A549 and H1650 cells was collected before and after treatment of RC, and transcriptomic sequencing was subsequently performed. Through the screening, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, Gene Ontology (GO) analysis, gene set enrichment analysis (GSEA) and protein-protein network interaction (PPI) analysis of differentially expressed genes (DEGs), the function enrichment and network construction of DEGs were carried out, and the selected key DEGs were finally verified by real-time quantitative polymerase chain reaction (qPCR). Results RC significantly altered the expression levels of 5,316 genes in A549 cells and 7,845 genes in H1650 cells. The most up-regulated genes were C11orf96 and RMRP, while the most down-regulated genes were ZBTB6, NUFIP1 and PTPN20. GSEA results showed that down-regulated genes were mainly related to cell cycle, DNA replication, RNA degradation and splicing pathways. KEGG analysis showed that down-regulated genes were enriched in cell cycle, spliceosome, and nucleocytoplasmic transport pathways. GO analysis showed that DEGs mainly involves in DNA replication, chromosome separation and non-coding RNA processing. PPI network analysis revealed hub genes, and qPCR confirmed that RC inhibits their expression levels. Conclusions RC inhibits the proliferation of A549 and H1650 cells via multiple signaling pathways and targets, where cell cycle-related genes, including CDK1, CDK2, CCNA2, CCNB1 and CDC6, are the core targets.