Transcriptomic Analysis on the Key Genes and Functional Pathways of Ramulus Cinnamomi in Repressing the Proliferation of Lung Cancer in vitro
DOI:
https://doi.org/10.62767/Keywords:
Ramulus Cinnamomi, non-small cell lung carcinoma, transcriptomics, epidermal growth factor receptor, gefitinibAbstract
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.
References
Zheng RS, Zhang SW, Sun KX, et al. Cancer statistics in China, 2016. Zhonghua Zhong Liu Za Zhi 2023; 45(3): 212-220.
Guo H, Li H, Zhu L, et al. "How long have I got?" in stage IV NSCLC patients with at least 3 months up to 10 years survival, accuracy of long-, intermediate-, and short-term survival prediction is not good enough to answer this question. Frontiers in Oncology 2021; 11: 761042.
Chen R, Zhao D, Wang LN. Mechanism and strategies on drug resistance of non-small cell lung cancer to EGFR-TKI. Cancer Research on Prevention and Treatment 2017; 44(03): 225-230.
Jackman D, Pao W, Riely GJ, et al. Clinical definition of acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small-cell lung cancer. Journal of Clinical Oncology 2010; 28(2): 357-360.
Qian CG, Jin L, Zhu LP, et al. Study on chemical constituents, antitumor and anti-inflammatory activities of Cinnamomi Ramulus. Chinese Traditional and Herbal Drugs 2022; 53(1): 31-40.
Liu J, Zhang Q, Li RL, et al. The traditional uses, phytochemistry, pharmacology and toxicology of Cinnamomi Ramulus: a review. Journal of Pharmacy and Pharmacology 2020; 72(3): 319-342.
Carvalho AA, Andrade LN, de Sousa ÉB, et al. Antitumor phenylpropanoids found in essential oils. BioMed Research International 2015; 2015: 392674.
Li J, Huang HY, Lin YC, et al. Cinnamomi Ramulus inhibits cancer cells growth by inducing G2/M arrest. Frontiers in Pharmacology 2023; 14: 1121799.
Pan B, Xia Y, Gao Z, et al. Cinnamomi Ramulus inhibits the growth of colon cancer cells via Akt/ERK signaling pathways. Chinese Medicine 2022; 17(1): 36.
Gong NJ, Liang H. Effects of Buyuan Decoction and Guizhi Fuling Pills on immune function of patients with middle and advanced non-small cell lung cancer. Anti-Tumor Pharmacy 2020; 10(2): 232-236.
Mei Z, Zhu FY, Ren Y, et al. Clinical experience on the application of modified Guizhi Fuling pills in tumors. World Latest Medicine Information 2018; 18(33): 137.
Wang XH, Yu MW, Wang XM. Analysis of WANG Xiao-min's treatment thoughts of malignant tumors with Chaihu Guizhi Decoction. Journal of Hubei University of Chinese Medicine 2023; 25(1): 116-118.
Huo XK, Jia YT. Overview of the clinical application of Huangqi Guizhi Wuwu Decoction in the treatment of tumors. Journal of Oncology in Chinese Medicine 2022; 4(3): 71-75.
Han L, Guo X, Bian H, et al. Guizhi Fuling Wan, a traditional Chinese herbal formula, sensitizes cisplatin-resistant human ovarian cancer cells through inactivation of the PI3K/AKT/mTOR pathway. Evidence-Based Complementary and Alternative Medicine 2016; 2016: 4651949.
Park GH, Song HM, Park SB, et al. Cytotoxic activity of the twigs of Cinnamomum cassia through the suppression of cell proliferation and the induction of apoptosis in human colorectal cancer cells. BMC Complementary Medicine and Therapies 2018;18(1): 28.
Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biology 2014; 15(12): 550.
Yu G, Wang LG, Han Y, et al. clusterProfiler: an R package for comparing biological themes among gene clusters. OMICS 2012; 16(5): 284-287.
Liberzon A, Birger C, Thorvaldsdóttir H, et al. The Molecular Signatures Database (MSigDB) hallmark gene set collection. Cell Systems 2015; 1(6): 417-425.
Szklarczyk D, Gable AL, Lyon D, et al. STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Research 2019; 47(D1): D607-D613.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 2001; 25(4): 402-408.
Hsu WH, Yang JC, Mok TS, et al. Overview of current systemic management of EGFR-mutant NSCLC. Annals of Oncology 2018; 29(suppl_1): i3-i9.
Sigismund S, Avanzato D, Lanzetti L. Emerging functions of the EGFR in cancer. Molecular Oncology 2018; 12(1): 3-20.
Shigematsu H, Lin L, Takahashi T, et al. Clinical and biological features associated with epidermal growth factor receptor gene mutations in lung cancers. Journal of the National Cancer Institute 2005; 97(5): 339-346.
Sun PD, Sun D. Mechanisms of Ramulus Cinnamomi water extract in inducing cell cycle progression and apoptosis of human colorectal cancer cells SW480. Chinese Journal of Gerontology 2020; 40(20): 4396-4399.
Jiang SH, Li L, Wu YS, et al. Mechanism of Guizhi Fuling Wan in inhibiting proliferation of human breast cancer MCF-7 cells. Chinese Journal of Experimental Traditional Medical Formulae 2018; 24(15): 132-136.
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The analyzed data sets generated during the study are available from the corresponding author on reasonable request.Issue
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