The synergistic antitumour effect of multi-components from Pulsatilla chinensis saponins in NCI-H460 lung cancer cell line through induction of apoptosis

Abstract

Context: Pulsatilla chinensis (Bunge) Regel (Ranunculaceae) possess antitumour effects; however, its antitumour potential has not been extensively investigated. Objective: To investigate the synergetic effect of multi-components from P. chinensis induced cell apoptosis and explore the mechanism. Materials and methods: The cytotoxicity was measured against NCI-H460, SMMC-7721, HCT-116 and U251 cell lines treated with eight monomers from P. chinensis. The synergetic effect of a combination of Pulsatilla saponin D (PSD), Raddeanoside R13 (R13), and Pulsatilla saponin A (PSA) was assessed using CalcuSyn3.0. Annexin V-FITC/PI and DAPI staining analyzed apoptosis of NCI-H460 cells treated with PSD, R13 and PSA alone or in combination. Proteins differential expression was analyzed using proteomic, DAVID Bioinformatics Resources, R software environment and KEGG database, and verified by western blotting. Results: PSD, R13, and PSA displayed greater antitumor activity with IC 50 values of 5.6, 5.1 and 10.5 mM against NCI-H460 cells compared with other monomers. The combination of PSD, R13, and PSA had a synergistic effect at CI ¼ 0.27 and induced 17.53% cells apoptotic detected by flow cytometric. Bioinformatic analysis showed an overview of the differentially expressed proteins and some signalling pathways. Moreover, some candidate proteins (LDHA, PI3K, NOL3 and cleaved-caspase-3) were validated by western blotting. Discussion and Conclusion: These results show PSD, R13, and PSA are good candidates as natural products for use in the treatment of lung cancer. Potential signalling pathways and protein targets need to be further validated. The application of the drug combination approach also provides a therapeutic strategy for cancer.

Anti-Proliferative and Pro-Apoptotic Activities of Hederacolchiside A1 On MCF-7 Cells Via ROS Regulation and JAK2/STAT3 Inactivation

“…Apoptosis is accompanied by changes in cell morphology, such as cell volume reduction, nuclear membrane shrinkage, chromatin concentration and aggregation, and apoptotic bodies (Zhang et al 2015). HA1 has a signi cant inhibitory effect on many types of tumor cell lines (NCI-H460, ZR75-1, HL-60, SMMC-7721, Bel-7402, MCF-7) by inducing apoptosis (Gerkens et al 2007;Liang et al 2016;Wang et al 2018;Guan et al 2020). HA1 can reportedly induce the apoptosis of breast cancer cells (ZR75-1 and MCF-7) and increase the expression of cleaved PARP and caspase-3 (Liang et al 2016).…”

“…The molecular mechanisms of the antitumor effects of HA1 remain uncertain, except for its reported membrane damage and high a nity for melanin (Delmas et al 2000;Debiton et al 2004). To date, research on the molecular mechanism is mainly limited to the inhibition of the PI3K/AKT/mTOR signaling pathway (Wang et al 2018;Guan et al 2020;Zhang et al 2020). It has been shown that STAT3 is crucial in numerous tumorigenesis processes, and activation of the JAK-STAT pathway plays a crucial role in the antiproliferation and apoptosis induction of breast cancer cells (Sansone et al 2012;Ma et al 2020).…”

“…Oleanane-type saponins, such as -hederin, β-hederin, hederacolchiside A1 (HA1), and Pulsatilla saponin A/D, have received considerable attention because of their strong antitumor activity in vitro and in vivo (Wang et al 2017;Guan et al 2020). HA1, as the main active ingredient, widely exists in the genera Anemone L. and Hedera L. These include Pulsatilla chinensis (Bunge) Regel, Anemone raddeana Regel, and Anemone accida Fr.…”

α-Hederin inhibits the growth of lung cancer A549 cells in vitro and in vivo by decreasing SIRT6 dependent glycolysis

“…In our previous study, we demonstrated that three monomers of Pulsatilla chinensis , PSA, R13, and PSD, can considerably inhibit the proliferation of tumour cell lines (Guan et al. 2020 ). α-Hederin ( Figure 1(a) ) is a monosaccharide chain pentacyclic triterpenoid saponin and is one of the effective components of the traditional Chinese medicine Pulsatilla chinensis .…”

“…Pulsatilla chinensis (Bunge) Regel (Ranunculaceae), which is used for its cooling and detoxifying abilities, has been reported to have many pharmacological characteristics such as being antimalarial and antibacterial (Cheng et al 2008) and an anticancer agent (Liu et al 2013). In our previous study, we demonstrated that three monomers of Pulsatilla chinensis, PSA, R13, and PSD, can considerably inhibit the proliferation of tumour cell lines (Guan et al 2020). a-Hederin (Figure 1(a)) is a monosaccharide chain pentacyclic triterpenoid saponin and is one of the effective components of the traditional Chinese medicine Pulsatilla chinensis.…”