[关键词]
[摘要]
研究辣木籽异硫氰酸酯(MITC)对皮肤鳞状细胞癌A431细胞生长的影响。通过MTT法、克隆形成实验及流式细胞术检测经不同浓度MITC处理后的A431细胞增殖、凋亡和细胞周期的变化情况;采用异种移植瘤模型检测MITC对A431细胞体内生长的影响。研究结果表明,随着MITC浓度和处理时间增加,A431细胞活力逐渐降低,在12 μM的MITC刺激72 h后,细胞存活率降低为10%(p<0.001);克隆形成实验表明,MITC(5和10 μM)显著抑制A431细胞克隆形成,24 h的细胞克隆形成率分别从100%降低到63.22%(p<0.001)和26.07%(p<0.001);细胞凋亡和周期检测结果表明,当MITC处理细胞48 h后,细胞凋亡率分别从17.22%增加到31.73%(p<0.001)和44.77%(p<0.001),S期细胞从7.43%显著增加到14.44%(p<0.001)和17.43%(p<0.001)。体内实验结果表明,用MITC干预异种移植瘤小鼠20天后,小鼠肿瘤体积从1549.02 mm3降低到857.77 mm3(p<0.05);肿瘤重量从1.30 g降低到0.91 g(p<0.05)。以上结果表明,MITC在体外和体内均能够抑制A431细胞的生长。
[Key word]
[Abstract]
The aim of this study was to investigate the effect of Moringa oleifera seed isothiocyanate (MITC) on the growth of skin squamous cell carcinoma cells (A431 cells). Following the treatment with different concentrations of MITC, the proliferation, apoptosis and cell cycle changes of A431 cells were detected by the MTT assay, clone formation experiment and flow cytometry; The xenograft tumor model was used to detect the effect of MITC on the growth of A431 cells in vivo. The results showed that the viability of A431 cells decreased gradually with the increase of MITC concentrations and processing time, and after a 72 h stimulation with MITC at 12 μM, the cells survival rate decreased to 10% (p<0.001); The results of the clonal formation experiments showed that MITC (5 and 10 μM) significantly inhibited the clonal formation of A431 cells, and the clonal formation rate after a 24 h stimulation with MITC decreased from 100% to 63.22% (p<0.001) and 26.07% (p<0.001); The results of apoptosis and cell cycle examinations showed that after a 48 h treatment with MITC, the apoptosis rate of A431 cells increased from 17.22% to 31.73% (p<0.001) and 44.77% (p<0.001), and the S-phase cells increased significantly from 7.43% to 14.44% (p<0.001) and 17.43% (p<0.001). The in vivo experiments showed that, after 20 days of MITC intervention in the mice with xenograft tumor, the tumor volume of the mice decreased from 1549.02 mm3 to 857.77 mm3 (p<0.05); The tumor weight decreased from 1.30 g to 0.91 g (p<0.05). The above results show that MITC can inhibit the growth of A431 cells in vitro and in vivo.
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[基金项目]
热作技术试验示范与服务支持—云南特色热带作物精深加工产品研发及示范(18190001);国家木薯产业技术体系(CARS-11-YNSJ);云南省中青年学术和技术带头人后备人才(2018HB040)