aaptamine 与顺铂联合用药对人肺腺癌顺铂耐药A549/DDP细胞的协同抑制作用

doi:10.13481/j.1671-587x.20210102

Abstract

Abstract: Objective

To investigate the synergetic inhibitory effect of aaptamine and cisplatin （DDP） on the DDP resistant human lung cancer A549/DDP cells， and to clarify its possible molecular mechanism.

Methods

The A549/DDP cells in logarithmic phase were selected and cultured with different concentrations of aaptamine and DDP for 48 h； the medium inhibitory concentrations（IC₅₀） of aaptamine and DDP were detected by CCK-8 assay； the combination index （CI） of aaptamine and DDP were analyzed by Chou-Talalay method and the optimal combination concentrations were confirmed. The cells were divided into control group， aaptamine group（5 mg·L^－1）， DDP group（1 mg·L^－1）， and combination group （5 mg·L^－1aaptamine +1 mg·L^－1DDP）. The proliferation activities of cells in various groups were detected by CCK-8 assay.The number of clones of the cells in various groups was determined by clonal formation assay. The migration abilities of the cells in various groups were detected by scratch assay. The apoptotic rates of cells in various groups were detected by double-staining flow cytometry. Western blotting method was performed to detect the expression levels of resistance-related proteins in the cells in various groups.

Results

The IC₅₀ values of aaptamine and DDP for the A549/DDP cells were 19.45 mg·L^－1 and 12.86 mg·L^－1， respectively. Co-treatment with aaptamine and DDP produced a synergistic effect at any selected concentrations in the A549/DDP cells. The proliferation assay results showed that compared with control group， aaptamine group and DDP group， the proliferation activity of the cells in combination group was decreased （P<0.01）. The clone formation assay results showed that compared with control group， aaptamine group and DDP group， the number of clones in combination group was decreased （P<0.05 or P<0.01）. The scratch assay results showed that compared with control group， aaptamine group and DDP group， the migration rate of cells in combination group was decreased significantly （P<0.01）. The flow cytometry results showed that compared with control group， aaptamine group and DDP group， the apoptotic rate of A549/DDP cells in combination group was markedly increased （P<0.05 or P<0.01）. The Western blotting results showed that compared with control group， aaptamine group and DDP group， the expression level of resistance related proteins ATP binding cassette subfamily G member 2（ABCG2） in the cells in combination group was decreased （P<0.05 or P<0.01），and the ratio of Bax/Bcl-2 was increased （P<0.05 or P<0.01）.Compared with control group，the expression level of excision repair cross complementing group 1（ERCC1） protein in combination group was decreased （P<0.05）.

Conclusion

Aaptamine has a synergistic inhibitory effect with DDP in the A549/DDP cells and its mechanism may be related to inhibiting proliferation， inducing apoptosis and decreasing the expression levels of resistance-related proteins ABCG2 and ERCC1.

Key words: aaptamine, cisplatin resistance, lung neoplasms, A549/DDP cells, synergistic effect

CLC Number:

R734.2

Shuang MIAO,Na NI,Lijuan YANG,Yan WU,Xuelin LI,Hongliang DONG,Kaikai GONG. Synergistic inhibitory effect of aaptamine and cisplatin on cisplatin resistant human lung adenocarcinoma A549/DDP cells[J].Journal of Jilin University(Medicine Edition), 2021, 47(1): 8-15.

Figures/Tables 7

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References 27

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Group	ABCG2	ERCC1	Bax /Bcl-2
Control	1.09±0.04	1.38±0.21	0.30±0.06
Aaptamine	1.04±0.08	1.16±0.10	0.66±0.18
DDP	0.90±0.02	0.92±0.19	0.63±0.13
Combination	0.59±0.08^*?#	0.55±0.04^*	1.74±0.05^**??##

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Synergistic inhibitory effect of aaptamine and cisplatin on cisplatin resistant human lung adenocarcinoma A549/DDP cells

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