虫草素联合阿霉素调控肾癌ACHN细胞系凋亡迁移的机制探讨  

Mechanism of cordycepin combined with adriamycin on apoptosis of renal carcinoma ACHN cell line

作　　者：张明 陈志云 霍玉杰 王沙凝 李再巧 王宣军[2] 盛军[2,4] 桂明英 马啸 ZHANG Ming;CHEN Zhiyun;HUO Yujie;WANG Shaning;LI Zaiqiao;WANG Xuanjun;SHENG Jun;GUI Mingying;MA Xiao(College of Food Science and Technology,Yunnan Agricultural University,Yunnan Kunming 650201,China;Key Laboratory of Pu'er Tea Science,Ministry of Education,Yunnan Agricultural University,Yunnan Kunming 650201,China;College of Longrun Pu'er Tea,Yunnan Agricultural University,Yunnan Kunming 650201,China;Yunnan Plateau Characteristic Agricultural Industry Research Institute,Yunnan Kunming 650201,China)

机构地区：[1]云南农业大学食品科学技术学院,云南昆明650201 [2]云南农业大学普洱茶学教育部重点实验室,云南昆明650201 [3]云南农业大学龙润普洱茶学院,云南昆明650201 [4]云南省高原特色农业产业研究院,云南昆明650201

出　　处：《现代肿瘤医学》2021年第1期9-15,共7页Journal of Modern Oncology

基　　金：云南省重大科技专项-绿色食品国际合作研究中心项目(编号:2019ZG009-06);云南农业大学自然科学青年基金(编号:A2006099)。

摘　　要：目的:探究肾癌ACHN细胞系经阿霉素组、阿霉素与虫草素联用组以及虫草素组刺激后,对细胞凋亡及迁移的影响及其相关机制探讨。方法:体外培养肾癌ACHN细胞系,采用MTT法检验不同浓度阿霉素组、阿霉素与虫草素联用组、虫草素组处理肾癌ACHN细胞系后细胞存活率;通过细胞划痕实验观察不同处理组肾癌ACHN细胞系的细胞迁移情况,形态学观察细胞外部形态,用HE染色和DAPI染色察看细胞内部变化和细胞核数量;并采用Western blot方法检测细胞内Bcl-2、Bax、MMP-2、MMP-9蛋白的表达情况。结果:与阿霉素高剂量组相比,联用组能够显著抑制肾癌ACHN细胞迁移,促进凋亡;并且发现减少细胞形态出现异常,HE和DAPI染色观察发现癌细胞数量明显减少,细胞核体积变大比率明显增加,细胞质中荧光强度减弱;Bax、Bcl-2、MMP-2蛋白表达差异显著。结论:虫草素与阿霉素联用组能够与阿霉素高剂量组达到同样抑制肾癌ACHN细胞迁移,并且显著促进凋亡蛋白基因表达的作用。本结果为天然植物有效成分与化学合成类药物联用治疗癌症提供了参考,为降低抗癌药物长时间耐药性提供了新途径。Objective:To explore the effects of apoptosis and migration of renal cell carcinoma ACHN cell line after stimulation with adriamycin,adriamycin and cordycepin,and cordycepin,and related mechanisms.Methods:In vitro culture of renal cell carcinoma ACHN cell line,MTT assay detected the cell viability of different concentrations of adriamycin group,adriamycin combined with cordycepin group,cordycepin group for treatment of renal cell carcinoma ACHN cell line.Cell scratch test observd the migration of different treatment groups for renal cell carcinoma ACHN cell line.External cell morphology was observed by morphological observation.The internal changes and the number of nuclei were observed by HE staining and DAPI staining.The intracellular Bcl-2,Bax,MMP-2,MMP-9 protein expression were detected by Western blot.Results:Compared with the high dose group of adriamycin,the combination group can significantly inhibit the migration of renal cell carcinoma ACHN cell line and promote apoptosis,and found that the cell morphology was abnormally reduced,and the number of cancer cells was significantly decreased by HE and DAPI staining,and the nuclear volume was observed.The ratio of enlargement was significantly increased.The fluorescence intensity in the cytoplasm was decreased.The expression of Bax,Bcl-2 and MMP-2 protein were significant difference.Conclusion:The combination of cordycepin and adriamycin can achieve the same inhibition of renal cancer ACHN cell line migration as the high-dose group,and significantly promote the expression of apoptotic protein gene,which results in the combination of natural plant active ingredients and chemical synthetic drugs to treat cancer.A certain theoretical reference provides a new way to reduce the long-term drug resistance of anticancer drugs.

关 键 词：虫草素 阿霉素 肾癌ACHN细胞系 凋亡 迁移 机制 

分 类 号：R737.12[医药卫生—肿瘤]