摘要
目的:探讨白杨素增强顺铂抗肿瘤作用的最佳条件并筛选敏感细胞。方法:设立阴性对照组、空白调零组及40μmol/L白杨素、5.0μg/mL顺铂、40μmol/L白杨素+5.0μg/mL顺铂3个处理组,选取人结直肠癌细胞(HCT-116)、人鼻咽癌细胞(CNE1)、人肝癌细胞(HepG2)、人胃癌细胞(BGC-823)分别作用24 h。采用四甲基噻唑蓝(MTT)法测定肿瘤细胞的增殖抑制率,采用Hoechst33342荧光染色法测定诱导肿瘤细胞的凋亡率,通过抑制率和凋亡率筛选出敏感细胞。采用L_9(3~3)正交设计方法,设立3个白杨素水平(10、20、40μmol/L),3个顺铂水平(13、2.5、5.0μg/mL),以及3种作用时间(12、74、36 h)处理敏感细胞,确定白杨素增强顺铂抗肿瘤作用的最佳作用条件。结果:与白杨素或顺铂单独处理组相比,MTT试验结果表明,白杨素联合顺铂作用组对上述4种肿瘤细胞增殖的抑制率均明显增加,差异均具有统计学意义(P均<0.01);Hoechst 33342染色实验发现,白杨素联合顺铂作用组诱导各肿瘤细胞的凋亡率亦均明显增加(P均<0.01)。白杨素联合顺铂作用组对人胃癌细胞BGC-823的增殖抑制率为(78.0±2.0)%、诱导细胞的凋亡率为(72.3±6.5)%。在4种细胞中,人胃癌细胞株BGC-823对白杨素联合顺铂抗肿瘤作用最敏感。正交实验的体外模型结果表明,最佳作用条件为白杨素40μmol/L联合顺铂5.0μg,mL,作用时间24 h。结论:白杨素能增强顺铂抑制人肿瘤细胞增殖和诱导人肿瘤细胞凋亡的作用。人胃癌细胞BGC-823是白杨素和顺铂联合作用的敏感细胞株。
OBJECTIVE： To study the anti-tumor effect of cisplatin by chrysin （ChR） in vitro. METHODS： Cultured cells from human colorectal cancer （HCT-116）,gastric cancer （BGC-823）,nasopharyngeal carcinoma （CNE1） and liver cancer （HepG2） were treated by chrysin （40 μmol/L）,cisplatin （5.0 μg/mL）,and chrysin （40 μmol/L）＋cisplatin （5.0 μg/mL） separately for 24 hours. The inhibitory rate of tumor cells was determined by MTT assay. The apoptosis rate of tumor cells was determined by Hoechst 33342 fluorescence staining. The sensitive cell line was identified by the inhibition rate and apoptosis index. The L9（33） orthogonal design of three levels of chrysin （10,20,40 μmol/L）,three levels of cisplatin （1.3,2.5,5.0 μg/mL）,and three time-points （12,24,36 h） was used to screen for the best combination effect of chrysin and cisplatin in the sensitive cells. RESULTS： Among all cell lines,the MTT assay showed that the inhibition rates from exposure to the combination of chrysin and cisplatin were significantly higher than that of the single treatment groups of chrysin and cisplatin （P 〈 0.01）. Similarly,Hoechst 33342 staining showed that the apoptosis rates from the combined treatment were significantly higher than those from the single treatment groups （P 〈 0.01）. From the combined treatment,the inhibitory rate in BGC-823 was （78.0±2.0）% and the apoptosis index was （72.3±6.5）%,indicating that this was the most sensitive among all tested cell lines. The results of in vitro orthogonal experiment showed that the most effective conditions were：40 μmol/L chrysin,5.0 μg/mL cisplatin,and 24 hours of treatment time. CONCLUSION： Chrysin enhanced the inhibitory effect of cisplatin via inhibition of cell proliferation and promotion of apoptosis in human tumor cell lines. Among the tested cell lines,the human gastric cancer cell line BGC-823 was the sensitive one to the combined effect of chrysin and cisplatin.
作者
杨美玲
李欣
黄俊明
张紫虹
易资梅
古梅英
黄健康
何云
YANG Meiling LI Xin HUANG Junming ZHANG Zihong YI Zimei GU Meiying HUANG Jiankang HE Yun(School of Public Health, Sun Yat-sen University, Guangzhou 510080 Guangdong Province Center for Disease Control and Prenention, Guangzhou 511430, Guangdong, China)
出处
《癌变．畸变．突变》
CAS
CSCD
2017年第1期37-41,共5页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
国家自然科学基金资助项目(81202199)
广东省医学科研基金(A2015055)
关键词
白杨素
顺铂
正交实验设计
抗肿瘤
ehrysin
eisplatin
orthogonal experiment design
anti-tumor
