Pristimerin enhances cisplatin-induced apoptosis in nasopharyngeal carcinoma cells via ROS-mediated deactivation of the PI3K/AKT signaling pathway

doi:10.12122/j.issn.1673-4254.2024.05.12

Abstract

Abstract:

Objective To explore the effect of pristimerin combined with cisplatin on proliferation and apoptosis of nasopharyngeal carcinoma cells. Methods CCK-8 assay was used to examine the survival rate of HNE-1 and CNE-2Z cells following treatment for 24 h with different concentrations of pristimerin, cisplatin or their combination. The changes in colony formation ability, apoptosis, and intracellular reactive oxygen species (ROS) levels of the treated cells were analyzed using colony formation assay and flow cytometry. Western blotting was performed to detect the changes in protein expressions in the cells. The effects of pre-treatment with NAC on proliferation, apoptosis, and PI3K/AKT signaling pathway were observed in pristimerin- and/or cisplatin-treated cells. Results Both pristimerin and cisplatin significantly lowered the survival rate of HNE-1 and CNE-2Z cells (P<0.05). Compared with pristimerin or cisplatin alone, their combination more strongly inhibited survival and colony formation ability of the cells, increased cell apoptosis rate and intracellular ROS levels, upregulated the protein expressions of Bax, cleaved caspase-3, and cleaved PARP, and downregulated the protein expressions of Bcl-2, Mcl-1, PARP and p-PI3K and p-AKT (P<0.05). NAC pretreatment significantly attenuated proliferation inhibition and apoptosis-promoting effects of pristimerin combined with cisplatin, and partially restored the downregulated protein expressions of p-PI3K and p-AKT in HNE-1 and CNE-2Z cells with the combined treatment (P<0.05). Conclusion Pristimerin can enhance cisplatin-induced proliferation inhibition and apoptosis in nasopharyngeal carcinoma cells, the mechanism of which may involve ROS-mediated deactivation of the PI3K/AKT signaling pathway.

Key words: nasopharyngeal carcinoma, pristimerin, cisplatin, reactive oxygen species, PI3K/AKT signaling pathway

Yuanyuan WANG, Teng CHEN, Xiaofan CONG, Yiran LI, Rui CHEN, Pei ZHANG, Xiaojin SUN, Surong ZHAO. Pristimerin enhances cisplatin-induced apoptosis in nasopharyngeal carcinoma cells via ROS-mediated deactivation of the PI3K/AKT signaling pathway[J]. Journal of Southern Medical University, 2024, 44(5): 904-912.

Figures/Tables 6

Fig.1 Effect of pristimerin (Pri) and/or cisplatin (DDP) on viability of HNE-1 and CNE-2Z cells. A: CCK-8 assay for detecting viability of HNE-1 and CNE-2Z cells with Pri treatment. B: CCK-8 assay for detecting viability of HNE-1 and CNE-2Z cells with DDP treatment. C: CCK-8 assay for detecting viability of HNE-1 and CNE-2Z cells treated with Pri, DDP or their combination. *P<0.05, **P<0.01 vs Ctrl group, ##P<0.01.

Fig.2 Effect of Pri combined with DDP on colony formation in HNE-1 and CNE-2Z cells. *P<0.05, **P<0.01 vs Ctrl group; #P<0.05, ##P<0.01 vs Pri or DDP group.

Fig.3 Effect of Pri combined with DDP on apoptosis of HNE-1 and CNE-2Z cells. A: Apoptosis of HNE-1 and CNE-2Z cells treated with Pri and/or DDP for 24 h detected by flow cytometry. B: Expressions of Bax, Bcl-2, Mcl-1, cleaved caspase-3 and cleaved PARP in HNE-1 cells detected by Western blotting. C: Expressions of Bax, Bcl-2, Mcl-1, cleaved caspase-3 and cleaved PARP in CNE-2Z cells detected by Western blotting. *P<0.05, **P<0.01 vs Ctrl group; #P<0.05, ##P<0.01 vs Pri or DDP group.

Fig.4 Effect of Pri combined with DDP on ROS levels in HNE-1 and CNE-2Z cells. A: ROS level of HNE-1 cells treated with Pri or DDP or their combination for 24 h detected by flow cytometry. B: ROS level in CNE-2Z cells treated with Pri or DDP or their combination for 24 h detected by flow cytometry. **P<0.01 vs Ctrl group; ##P<0.01.

Fig.5 ROS was involved in mediating the effect of Pri combined with DDP on proliferation and apoptosis in HNE-1 and CNE-2Z cells. A: CCK-8 assay for detecting viability of HNE-1 and CNE-2Z cells treated with NAC followed by Pri combined with DDP. **P<0.01vs Ctrl group, ##P<0.01 vs Pri+DDP group. B, C: Protein levels of Bax, Bcl-2, Mcl-1, cleaved caspase-3 and cleaved PARP in HNE-1 cells and CNE-2Z cells detected by Western blotting. *P<0.05, **P<0.01 vs Pri+DDP group.

Fig.6 Effect of Pri combined with DDP on expression of PI3K/AKT signaling pathway-related proteins in HNE-1 and CNE-2Z cells. A, B: Protein expression levels of PI3K, AKT, p-PI3K and p-AKT in HNE-1 cells and CNE-2Z cells detected by Western blotting. *P<0.05, **P<0.01 vs Ctrl group; #P<0.05, ##P<0.01 vs Pri or DDP group. C, D: Western blotting of protein expression levels of PI3K, AKT, p-PI3K and p-AKT in HNE-1 cells and CNE-2Z cells with NAC pretreatment. *P<0.05 vs Pri+DDP group.

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