Identification of Phytochemicals and RNA-Seq Combined in Silico Analysis of Active Compounds and Lung Cancer Related Targets of Cuscuta chinensis Lam

Objectives Cuscuta chinensis Lam.(C. chinensis) is a common medicinal plant, rich in bioactive compounds. However, the anti-lung cancer active compounds of C. chinensis, and related molecular mechanisms are not clear yet. Methods The peaks of CLW in the fingerprinting were analyzed using LC/MS/QTOF. Total phenolic and flavonoid contents of CLW were evaluated utilizing Foline-Ciocalteu colorimetric and aluminum chloride assay. Cytotoxicity assay was performed for cell viability in vitro. Gene expression profiles were determined using RNA-seq analysis. The active compounds and potential targets were screened in silico. A herb-comound-target-disease network was constructed to prediction of key targets. Docking analysis of the active compounds with key targets were analyzed using Autodock Vina 1.1.2. Toxicity assessment was conducted using ToxTree v3.1.0. Results 32 peaks in fingerprinting of CLW were tentatively identified, and the main chemical ingredients of CLW were predicted as flavonoids, glycosidic acids, lignans, phenylpropanoids, phenolic acids and resin glycosides, and further quantified its phenolics and flavonoids. CLW inhibited viability of LLC cells in a time and dose dependent manner. Moreover, RNA-seq analysis revealed 79 genes expression were significantly altered in LLC cells under CLW treatment. In addition, Gene Ontology biological processes (GOBP) analysis suggested that these genes are significantly associated with regulation of protein function and cell proliferation. Furthermore, in silico screening results identified 33 active compounds in C. chinensis. Besides, a RNA-seq analysis combined with network pharmacology effectively identified 8 chemical compounds highly related to Bone morphogenetic protein 1 (BMP1), Cyclin C (CCNC) and Heat shock protein 90β1 (HSP90B1) in lung cancer. Finally, molecular docking results and toxicity assessment demonstrated that medioresinol, (-)-pinoresinol, gitoxigenin and n-trans-feruloyltyramine shows a strong binding affinity with BMP1B, CCNC and HSP90B1, and were potential non-toxic. Conclusions This results suggesting that C. chinensis may provides a new promising lead compounds for lung cancer treatment.