摘要
目的:采用黄芪多糖(APS)与放射治疗(IR)联用,研究APS对人鼻咽癌CNE^-1细胞的放疗敏感性及上皮间质转化(EMT)的作用机制。方法:采用细胞计数(CCK-8)法检测不同质量浓度APS(0,6.25,12.5,25,50,100,200 g·L^-1)对CNE^-1细胞的细胞毒性;克隆形成实验计算12.5 g·L^-1APS与不同放射剂量(0,2,4,6 Gy)联用后对CNE^-1细胞的存活分数(SF),利用线性二次方程数学模型(LQ)根据SF值绘制放射敏感曲线;细胞划痕和transwell小室实验分别检测各组细胞的迁移和侵袭能力;流式细胞仪检测各组细胞的凋亡情况;蛋白免疫印迹法(Western blot)检测各组细胞的EMT标记物、凋亡标记物以及蛋白激酶B/细胞外调节蛋白激酶(Akt/ERK)通路蛋白的表达水平。结果:克隆形成实验和放射敏感曲线结果表明,非毒性剂量12.5 g·L^-1APS与4 Gy放射剂量联合给药可以明显增加CNE^-1细胞的放疗敏感性;与空白组及IR组比较,APS与IR联用可以抑制CNE^-1细胞的迁移和侵袭能力(P<0.05),明显增加CNE^-1细胞的凋亡率(P<0.05)。与空白组及IR组比较,APS与IR联用可以显著下调间质型钙黏蛋白(N-cadherin),p-Akt和p-ERK蛋白表达水平,明显上调上皮型钙黏蛋白(E-cadherin),B淋巴细胞瘤-2相关X蛋白(Bax)和半胱天冬氨酸蛋白酶-3(Caspase-3)蛋白表达水平(P<0.05)。结论:APS与IR联用可以抑制CNE^-1细胞的迁移和侵袭,增加放疗引起的细胞凋亡,其可能通过抑制EMT和Akt/ERK通路有关。
Objective: Astragalus polysaccharide(APS)was used in combination with ionizing radiation(IR)to investigate the mechanism of APS on the radiosensitivity of human nasopharyngeal narcinoma CNE^-1 cells and the epithelial-mesenchymal transition(EMT). Method:Cell counting kit-8(CCK-8)was used to detect the cytotoxicity of different concentrations of APS(0,6.25,12.5,25,50,100,200 g·L^-1)on CNE^-1 cells. Colony formation assay was used to calculate the survival fraction(survival fraction,SF)of CNE^-1 cells treated with 12.5 g·L^-1 APS combined with different radiation doses(0,2,4,6 Gy). The linear quadratic equation mathematical model(LQ)was used to draw the radiosensitivity curve according to SF value. Cell scratch and transwell chamber test were used to detect the migration and invasion ability of cells in each group. The apoptosis of cells in each group was detected by flow cytometry,Western blot was used to detect the expressions of EMT markers,apoptosis markers and protein kinase B/extracellular regulated protein kinases(Akt/ERK)pathway proteins in each group. Result:The results of colony formation assay and radiosensitivity curve showed that the combination of non-toxic dose of 12.5 g·L^-1 APS and radiation dose of 4 Gy could significantly increase the radiosensitivity of CNE^-1 cells. Compared with blank group and IR group,APS combined with IR could significantly inhibit the migration and invasion of CNE^-1 cells(P<0.05),and increase the rate of apoptosis(P<0.05). In addition,compared with the blank group and the IR group,APS combined with IR could significantly down-regulate the expressions of N-cadherin,p-Akt and p-ERK,and significantly up-regulate the expressions of E-cadherin,Bax and Caspase-3(P<0.05). Conclusion: APS combined with IR can inhibit the migration and invasion of CNE^-1 cells,and increase the apoptosis induced by radiotherapy,which may be related to the inhibition of EMT and Akt/ERK pathway.
作者
张树聪
蔡治祥
王学涛
李志英
宋慧胜
ZHANG Shu-cong;CAI Zhi-xiang;WANG Xue-tao;LI Zhi-ying;SONG Hui-sheng(The Second Clinical School of Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Guangzhou Panyu Central Hospital,Guangzhou 511400,China;The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Qingyuan People's Hospital,The Sixth Affiliated Hospital of Guangzhou Medical University,Qingyuan 511518,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第20期59-66,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
广东省自筹经费类科研计划项目(2017ZC0165)。
关键词
黄芪多糖
CNE-1细胞
放疗增敏
上皮间质转化
迁移
细胞凋亡
astragalus polysaccharides
CNE-1 cells
radiosensitization
epithelial-mesenchymal transition
migration
apoptosis
