扁蒴藤素通过活性氧调控PI3K/AKT通路增强顺铂诱导鼻咽癌细胞凋亡

doi:10.12122/j.issn.1673-4254.2024.05.12

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (5): 904-912.doi: 10.12122/j.issn.1673-4254.2024.05.12

扁蒴藤素通过活性氧调控PI3K/AKT通路增强顺铂诱导鼻咽癌细胞凋亡

王媛媛(), 陈腾, 从小凡, 李依然, 陈蕊, 张配, 孙小锦(), 赵素容()

蚌埠医科大学药学院//安徽省生化药物工程技术研究中心，安徽蚌埠 233030

收稿日期:2023-12-20 出版日期:2024-05-20 发布日期:2024-06-06
通讯作者: 孙小锦,赵素容 E-mail:wangyy0424@163.com;aijosxj@163.com;inwindangel@qq.com
作者简介:王媛媛，硕士，E-mail: wangyy0424@163.com
基金资助:
国家自然科学基金(81603155);安徽省高校自然科学研究重点项目(KJ2021A0736);蚌埠医学院自然科学重点项目(2021byzd018);安徽省大学生创新创业训练项目(S202210367013);蚌埠医学院研究生科研创新计划自然科学项目(Byycx22061)

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

Yuanyuan WANG(), Teng CHEN, Xiaofan CONG, Yiran LI, Rui CHEN, Pei ZHANG, Xiaojin SUN(), Surong ZHAO()

School of Pharmacy, Bengbu Medical University//Anhui Provincal Engineering Technology Research Center of Biochemical Pharmaceuticals, Bengbu 233030, China

Received:2023-12-20 Online:2024-05-20 Published:2024-06-06
Contact: Xiaojin SUN, Surong ZHAO E-mail:wangyy0424@163.com;aijosxj@163.com;inwindangel@qq.com
Supported by:
National Natural Science Foundation of China(81603155)

摘要/Abstract

摘要：

目的观察扁蒴藤素和顺铂对鼻咽癌细胞增殖和凋亡的影响，并探讨其作用机制。方法 CCK-8法检测不同浓度扁蒴藤素、顺铂处理24 h后HNE-1和CNE-2Z细胞的存活率，集落形成实验观察细胞集落形成能力，流式细胞术检测细胞凋亡和活性氧（ROS）水平，Western blotting检测蛋白表达。N-乙酰半胱氨酸（NAC）预处理后，检测细胞增殖、凋亡及通路的影响。结果 CCK-8结果显示，扁蒴藤素和顺铂均可抑制HNE-1和CNE-2Z细胞的存活率（P<0.05）。与单用扁蒴藤素或顺铂相比，联合使用明显抑制细胞存活率和细胞集落形成能力（P<0.05），升高细胞凋亡率（P<0.01）和细胞内ROS水平（P<0.01），上调Bax、Cleaved caspase-3和Cleaved PARP的表达（P<0.05），下调Bcl-2、Mcl-1和PARP的表达（P<0.05），同时，下调细胞中p-PI3K和p-AKT的表达（P<0.05）。而NAC可部分逆转扁蒴藤素联合顺铂对HNE-1和CNE-2Z细胞的增殖抑制（P<0.01）和凋亡诱导作用（P<0.05），部分恢复p-PI3K和p-AKT的下调作用（P<0.05）。结论扁蒴藤素能增强鼻咽癌细胞对顺铂的增殖抑制和凋亡诱导作用，其机制可能与ROS介导的PI3K/AKT信号通路失活有关。

关键词: 鼻咽癌, 扁朔藤素, 顺铂, 活性氧, PI3K/AKT信号通路

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

王媛媛, 陈腾, 从小凡, 李依然, 陈蕊, 张配, 孙小锦, 赵素容. 扁蒴藤素通过活性氧调控PI3K/AKT通路增强顺铂诱导鼻咽癌细胞凋亡[J]. 南方医科大学学报, 2024, 44(5): 904-912.

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.

图/表 6

图1 扁蒴藤素和/或顺铂对鼻咽癌细胞活力的影响

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.

图2 扁蒴藤素联合顺铂对鼻咽癌细胞集落形成的影响

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.

图3 扁蒴藤素联合顺铂对鼻咽癌细胞凋亡的影响

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.

图4 扁蒴藤素联合顺铂对鼻咽癌细胞ROS水平的影响

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.

图5 ROS参与扁蒴藤素联合顺铂对鼻咽癌细胞增殖和凋亡的影响

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.

图6 扁蒴藤素联合顺铂对PI3K/AKT信号通路相关蛋白表达的影响

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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扁蒴藤素通过活性氧调控PI3K/AKT通路增强顺铂诱导鼻咽癌细胞凋亡

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

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