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目的:研究茱萸碱对涎腺腺样囊性癌SACC-M细胞生长的调控影响,为临床治疗涎腺腺样囊性癌提供理论依据。方法:选择口腔颌面外科手术治疗的20例涎腺腺样囊性癌患者的癌组织细胞ACC-M,置于含有RPMI1640、小牛血清、青霉素及链霉素的培养基中,恒温37℃下传代培养。再使用MTT比色实验将对数生长期的细胞悬浮液种于孔板内培养24h后,按浓度由小到大5种不同浓度比例加入吴茱萸碱溶液,另外设1组空白对照,1组调零组,进行培养后分别加入MTT、二甲基亚砜溶液用酶标法测定各组吸光度值并计算细胞生长的抑制率,显微镜下观察细胞生长状况,用流式细胞仪检测细胞凋亡状况。结果:MTT比色试验结果显示,吴茱萸碱对SACC-M细胞的生长具有明显的抑制效果,它的抑制率随时间的延长而增长,在药物作用后的24、48、72h的抑制率分别为35%,65%,85%,经过对细胞的抑制作用与药物浓度做二变量相关性分析可知二者呈正相关性(r=1.194,r<0.01)。显微镜下观察可见对照组细胞生长迅速,实验组生长缓慢;流式细胞仪下可见实验组大量细胞死亡,对照组凋亡率低。结论:吴茱萸碱对涎腺腺样囊性癌细胞具有生长抑制作用,它可以控制SACC-M细胞的增殖与凋亡,且具有随浓度升高药效增强的趋势,可以将此运用于SACC的治疗。
Objective: To study the regulation of the evodiamine on the growth of salivary adenoid cystic carcinoma SACC-M cells and to provide a theoretical basis for clinical treatment of salivary adenoid cystic carcinoma. METHODS: ACC-M cells from 20 patients with salivary adenoid cystic carcinoma who underwent oral and maxillofacial surgery were selected and cultured in a medium containing RPMI1640, fetal bovine serum, penicillin and streptomycin and incubated at 37 ℃ Subculture MTT colorimetric assay was then used to culture the cell suspension in logarithmic growth phase in the well plate for 24 h. Five concentrations of evodiamine solution were added in ascending order of concentration, and one blank control group and one set of control group After the culture, MTT was added respectively. The absorbance value of each group was measured by enzyme-labeled method and the inhibition rate of cell growth was calculated. The cell growth was observed under the microscope. The apoptosis status was detected by flow cytometry . Results: The results of MTT assay showed that evodiamine significantly inhibited the growth of SACC-M cells and its inhibitory rate increased with time. The inhibitory rates of evodiamine at 24, 48 and 72 h after treatment were 35%, 65% and 85% respectively. There was a positive correlation between the inhibitory effect on the cells and the drug concentration by two-variable correlation (r = 1.194, r <0.01). Under the microscope, the cells in the control group grew rapidly and the experimental group grew slowly. Under the flow cytometry, a large number of cells died in the experimental group and the apoptosis rate in the control group was low. CONCLUSION: Evodiamine can inhibit the growth of human salivary adenoid cystic carcinoma cells. It can control the proliferation and apoptosis of SACC-M cells and has the potential to increase with increasing concentration. This can be applied to SACC treatment.