华蟾素对人皮肤鳞状细胞癌A431细胞增殖、凋亡的影响及机制分析  

Effect and mechanism of cinobufacin on proliferation and apoptosis of human skin squamous cell carcinoma cell line A431

作　　者：朱小美 张小超 曹育春[1] 黄永初 Zhu Xiaomei;Zhang Xiaochao;Cao Yuchun;Huang Yongchu(Department of Dermatology,Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology,Wuhuan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院皮肤科,武汉430030

出　　处：《肿瘤综合治疗电子杂志》2022年第4期32-36,共5页Journal of Multidisciplinary Cancer Management(Electronic Version)

基　　金：国家自然科学基金面上项目(81974308)。

摘　　要：目的 探讨华蟾素对人皮肤鳞状细胞癌A431细胞增殖、凋亡的影响及机制。方法 采用CCK-8实验分析不同剂量华蟾素对A431细胞增殖的影响,并将A431细胞随机分为对照组、低剂量组(100 nmol/L华蟾素)、中等剂量组(500 nmol/L华蟾素)、高剂量组(1000 nmol/L华蟾素),通过流式细胞术(AnnexinⅤ-FITC)实验比较各组细胞凋亡比例,采用蛋白质印迹法实验比较各组细胞PCNA、Bax、Bcl-2、Akt、p-Akt、ERK、p-ERK表达水平。结果 CCK-8实验和流式细胞术实验表明华蟾素作用48 h后明显抑制了A431细胞增殖,诱导细胞凋亡,且该趋势与药物剂量有关,随着华蟾素的药物剂量增加,细胞活力逐渐降低,细胞凋亡比例逐渐升高(P<0.05)。蛋白质印迹法实验表明华蟾素作用48 h后未明显改变Akt、ERK的总蛋白表达水平,明显抑制了A431细胞PCNA、Bcl-2、p-Akt、p-ERK的表达水平,升高了Bax表达水平,且该趋势与药物剂量有关,随着华蟾素的药物剂量增加,A431细胞PCNA、Bcl-2、p-Akt、p-ERK的表达水平逐渐降低,Bax表达水平逐渐升高。结论 华蟾素能显著抑制人皮肤鳞癌细胞A431的增殖,诱导细胞凋亡,呈剂量依赖性,可能与抑制Akt和ERK蛋白磷酸化水平有关。Objective To investigate the effect and mechanism of cinobufacin on proliferation and apoptosis of human cutaneous squamous cell carcinoma cell line A431.Method CCK-8 experiment was used to analyze the effect of different doses of cinobufacin on the proliferation of A431 cells.A431 cells were randomly divided into control group,low-dose group (100 nmol/L cinobufacin),medium-dose group (500 nmol/L cinobufacin) and high-dose group (1000 nmol/L cinobufacin).The apoptosis ratio of each group was compared by flow cytometry (Annexin Ⅴ-FITC) experiment,and the expression levels of PCNA,Bax,Bcl-2,Akt,p-Akt,ERK and p-ERK were compared among all groups through Western blotting experiment.Result CCK-8 and flow cytometry experiment showed that cinobufacin significantly inhibited the proliferation of A431 cells and induced apoptosis after 48 h,and the trend was related to the drug dose.With the increase of cinobufacin concentration,the cell viability gradually decreased and the proportion of apoptosis gradually increased (P<0.05).Western blotting experiment showed that cinobufacin did not significantly change the total protein expression levels of Akt and ERK after 48 h,while it significantly inhibited the expression levels of PCNA,Bcl-2,p-Akt and p-ERK of A431 cells,and increased the expression level of Bax.The trend was related to the drug dose.With the increase of cinobufacin concentration,the expression levels of PCNA,Bcl-2,p-Akt and p-ERK of A431 cells gradually decreased,and the expression level of Bax gradually increased.Conclusion Cinobufacin can significantly inhibit the proliferation of human cutaneous squamous cell carcinoma cell line A431 and induce apoptosis in a dose-dependent manner,which may be related to the inhibition of Akt and ERK protein phosphorylation levels.

关 键 词：华蟾素 皮肤鳞状细胞癌 增殖 凋亡 AKT ERK 

分 类 号：R285[医药卫生—中药学]