BPPA通过活性氧介导线粒体损伤促进头颈癌细胞死亡的作用研究  

BPPA promotes head and neck cancer cell death via mitochondrial damage induced by reactive oxygen species

作　　者：曾叶茜 周莘卓 冯宇菲 唐艳 王勇德 胡春生 ZENG Ye-qian;ZHOU Xin-zhuo;FENG Yu-fei;TANG Yan;WANG Yong-de;HU Chun-sheng(College of Pharmacy,International Academy of Targeted Therapeutics and Innovation,,National&Local Joint Engineering Research Center of Targeted and Innovative Therapeutics,Chongqing Universityof Arts and Sciences,Chongqing 402160,China;Chongqing Key Laboratory of Chinese Medicine&Health Science,Chongqing Academy of Chinese Materia Medica,Chongqing 400065,China)

机构地区：[1]重庆文理学院药学院/创新靶向药物国际研究院,创新靶向药物国家地方联合工程中心,重庆402160 [2]重庆市中药研究院大健康创新中心,重庆400065

出　　处：《中国临床药理学杂志》2025年第3期340-344,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81902357);中国博士后科学基金资助项目(2022M710550)。

摘　　要：目的探讨新型吡唑[1,5-a]嘧啶衍生物(BPPA)对人舌鳞状癌细胞Cal33的抗肿瘤活性及作用机制。方法将Cal33细胞分为空白组(正常培养)、低、中、高剂量实验组(1、2.5、5μmol·L^(-1)BPPA)及联合组[2.5μmol·L^(-1)BPPA和5 mmol·L^(-1)N-乙酰-L-半胱氨酸(NAC)]。用噻唑蓝法检测BPPA对肿瘤细胞的存活率;用流式细胞术检测细胞的凋亡情况;用蛋白质印迹法检测凋亡相关蛋白相对表达水平;用免疫荧光分析活性氧及线粒体膜电位的变化情况。结果空白组、低、中、高剂量实验组及联合组的Cal33细胞存活率分别为(99.56±0.91)%、(58.31±2.31)%、(47.93±1.67)%、(29.35±4.11)%和(70.27±1.21)%,凋亡率分别为(8.07±1.01)%、(44.04±0.94)%、(49.85±1.75)%、(66.79±0.83)%和(24.33±1.04)%;活性氧检测的相对荧光强度分别为1.23±0.21、3.37±0.35、15.53±1.46、20.28±1.24和5.59±0.52,线粒体膜电位红色荧光/绿色荧光值分别为4.69±0.11、4.24±0.12、0.86±0.16、0.46±0.05和2.99±0.11。低、中、高剂量实验组上述指标与空白组比较,联合组上述指标与中剂量实验组比较,在统计学上差异均有统计学意义(P<0.01,P<0.001)。空白组和低、中、高剂量实验组促凋亡蛋白B淋巴细胞瘤-2基因相关X蛋白(Bax)相对表达水平分别为1.00±0.02、1.94±0.03、3.73±0.06和4.64±0.08,同时抑制凋亡蛋白B淋巴细胞瘤-2基因蛋白(Bcl-2)的相对表达水平分别为1.00±0.02、0.75±0.04、0.62±0.02和0.46±0.03。低、中、高剂量实验组上述指标与空白组比较,在统计学上差异均有统计学意义(P<0.01,P<0.001)。结论化合物BPPA具有抗头颈癌活性,其机制是通过促进活性氧生成介导线粒体膜电位降低诱导细胞凋亡导致肿瘤死亡。Objective To investigate the antitumor activity and mechanism of a pyrazole[1,5-a]pyrimidine derivative(BPPA)on human tongue squamous cell carcinoma Cal33 cells.Methods Cal33 cells were divided into blank group(normal culture),experimental-L,-M,-H groups(1,2.5 and 5μmol·L^(-1)BPPA)and combined group[2.5μmol·L^(-1)BPPA and 5 mmol·L^(-1)N-acety-L-cysteine(NAC)].Cell viability was determined by thiazole blue assay.Cell apoptosis was measured using flow cytometry,while apoptosis-related protein expression levels were determined by Western blot analysis.Changes in reactive oxygen species and mitochondrial membrane potential were evaluated using immunofluorescence assay.Results The survival rates of Cal33 cells in blank,experimental-L,-M,-H and combined groups were(99.56±0.91)%,(58.31±2.31)%,(47.93±1.67)%,(29.35±4.11)%and(70.27±1.21)%,respectively;cell apoptotic rates were(8.07±1.01)%,(44.04±0.94)%,(49.85±1.75)%,(66.79±0.83)%and(24.33±1.04)%,respectively;the relative fluorescence intensities of reactive oxygen species(ROS)were 1.23±0.21,3.37±0.35,15.53±1.46,20.28±1.24 and 5.59±0.52,respectively;the mitochondrial membrane potential red fluorescence/green fluorescence values were 4.69±0.11,4.24±0.12,0.86±0.16,0.46±0.05 and 2.99±0.11,respectively.Compared with blank group,the above indexes in the experimental-L,-M,-H groups were statistically significant(P<0.01,P<0.001).Compared with experimental-M group,the above indexes in combined group were statistically significant(P<0.01,P<0.001).The relative levels of pro-apoptotic B-cell lymphoma-2(Bcl-2)-Associated X protein(Bax)in blank,experimental-L,-M,-H groups were 1.00±0.02,1.94±0.03,3.73±0.06 and4.64±0.08,respectively;the relative levels of anti-apoptotic protein Bcl-2 were 1.00±0.02,0.75±0.04,0.62±0.02 and 0.46±0.03,respectively.Compared with blank group,the above indexes in the experimental-L,-M,-H groups were statistically significant(P<0.01,P<0.001).Conclusion BPPA exhibits anti-head and neck cancer activity by ROS mediating the decreas

关 键 词：吡唑类衍生物 头颈癌 细胞凋亡 活性氧 线粒体膜电位 

分 类 号：R979.1[医药卫生—药品]