Marine sponge-derived smenospongine overcomes resistance of cisplatin via inhibiting EGFR-Akt-ABCG2 pathway in NSCLC cells

doi:10.3969/j.issn.1674-8115.2022.08.004

Abstract

Abstract:

Objective ·To explore the antitumor activities and mechanisms of the sesquiterpene amine quinone compound smenospongine (SME) in non-small cell lung cancer (NSCLC) cell line A549 and cisplatin-resistant cell line A549/DDP. Methods ·Parental cell A549 and its cisplatin-resistant cell A549/DDP were used as research models. CCK-8 assay was used to detect the half maximum inhibitory concentration (IC₅₀) of the two cell lines treated with first-line chemotherapeutic drugs, in order to verify the multidrug resistance of A549/DDP. Colony formation assay was used to detect the effect of SME on the proliferation of parental and drug-resistant cells. Transwell invasion assay and Western blotting were used to detect the effect of SME on epithelial to mesenchymal transformation (EMT) of A549/DDP; Western blotting and quantitative real-time PCR (qRT-PCR) were used to detect the protein and mRNA expression levels of multidrug resistance genes regulated by SME in two cell lines and the molecular mechanism of drug resistance. Furthermore, Western blotting was used to detect the effect of SME on the upstream protein of multidrug resistance protein, ATP-binding cassette superfamily G member 2 (ABCG2), and flow cytometry was used to detect the effect of SME on the cell cycle of parental and cisplatin-resistant cells. TdT-mediated dUTP Nick-end labeling (TUNEL) and Western blotting were used to detect the effect of SME on apoptosis of the two cell lines. Results ·Compared with parental cells, cisplatin-resistant A549/DDP cells showed significant resistance to cisplatin and showed multidrug resistance to the first-line chemotherapy drugs used on lung cancer. SME markedly inhibited the proliferation and clone formation of A549 and A549/DDP as well as the EMT of drug-resistant cell. SME notably down-regulated the expression of ABCG2 protein and mRNA, and inhibited the epidermal growth factor receptor (EGFR)-serine/threonine kinase (Akt) signal pathway upstream of ABCG2, which thereby down-regulated ABCG2, and positively regulated FoxO1. SME induced G0/G1 arrest and induced apoptosis of both cells. Conclusion ·As a new small molecular compound in overcoming the drug resistance of NSCLC, SME inhibits the cell viability of A549 and A549/DDP by restraining EGFR-Akt signal pathway which thereby down-regulates ABCG2, positively regulates FoxO1 and inhibits EMT of A549/DDP, which finally leads to apoptosis.

Key words: smenospongine (SME), non-small cell lung cancer (NSCLC), cisplatin resistance, ATP-binding cassette superfamily G member 2 (ABCG2), serine/threonine kinase (Akt)

CLC Number:

LIAO Yahui, LIU Liyun, ZHU Hongrui, LIN Houwen, YAN Jizhou, SUN Fan. Marine sponge-derived smenospongine overcomes resistance of cisplatin via inhibiting EGFR-Akt-ABCG2 pathway in NSCLC cells[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022, 42(8): 997-1007.

Figures/Tables 7

Fig 1 Authentication of cisplatin-resistant cells

Tab 1 MDR features of cisplatin-resistant cells

Chemical	IC₅₀/（μmol×L^-1）		RI
Chemical	A549	A549/DDP	RI
DDP	3.730±0.080	155.100±6.290	41.550
GEM	0.570±0.070	2.880±0.100	5.050
NVB	0.006±0.001	5.370±0.050	895.000
Pemetrexed	48.040±6.340	304.670±0.754	6.342
5-FU	31.630±0.930	33.710±0.100	1.070

Fig 2 SME inhibits cell proliferation and EMT in cisplatin-resistant cells

Fig 3 SME inhibits the mRNA expression of ABCG2 in cisplatin-resistant cells

Fig 4 SME inhibits the activity of EGFR-Akt pathway in cisplatin-resistant cells

Fig 5 SME induces G0/G1 arrest and apoptosis in cisplatin-resistant cells

Fig 6 A schematic diagram depicting a model of the inhibition of SME on the EGFR-Akt-ABCG2 pathway

TrendMD

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