摘要
目的探索组蛋白去甲基化酶JMJD3对乳腺癌干细胞（BCSCs）的影响。方法采用改良后的无血清悬浮培养肿瘤干细胞的方法从乳腺癌MCF-7细胞系中分离扩增BCSCs;通过流式侧群细胞分选和免疫缺陷裸鼠腋下接种的方法检测BCSCs的生物学特性和致瘤性。采用Western Blot研究MCF-7及BCSCs中JMJD3表达的差异;采用CCK-8法检测JMJD3促进剂U0126及抑制剂GSK-J1对MCF-7增殖的影响;通过流式细胞侧群分选,检测GSK-J1、U0126对MCF-7中BCSCs比例的影响;采用Western Blot法研究GSK-J1、U0126对BCSCs中JMJD3蛋白表达的影响。结果改良后的无血清悬浮培养方法可成功地从MCF-7细胞系中分离并富集BCSCs;与MCF-7比较,BCSCs呈现出高致瘤性、JMJD3蛋白低表达性;U0126可抑制MCF-7的增殖,并降低MCF-7中BCSCs的比例,促进BCSCs中JMJD3的表达,而GSK-J1则呈现出相反的作用。结论JMJD3可抑制乳腺癌细胞的增殖,这种抑制作用可能与减少MCF-7中干细胞的比例有关。
OBJECTIVE To explore the effects of a histone demethylase JMJD3 on the breast cancer stem cells（ BCSCs）.METHODS Serum-free suspension culture was used to optimize the traditional method to separate and expand BCSCs. Flow SP analysis to isolate SP cells was used to determine the percentage of cancer stem cells in MCF-7 spheres,and compare the tumorigenic potential of BCSCs and MCF-7 in immune-deficient mice. Western Blot was used to examine the different expression of JMJD3 in MCF-7 and BCSCs. CCK-8 assay was used to measure the effects of the JMJD3 promoter U0126 and inhibitor GSK-J1 on the proliferation of MCF-7. Flow SP analysis was used to analyse the stem cell proportion of MCF-7 after the treatment of U0126 and GSK-J1 compared with the control group. Western Blot was used to compare the effects of GSK-J1 and U0126 on the expression of JMJD3 in BCSCs. RESULTS BCSCs were successfully separated and expanded from MCF-7 by the optimized method. Compared with MCF-7,BCSCs showed high tumorigenicity and low JMJD3 protein levels. U0126 inhibited the proliferation of MCF-7 and reduced the proportion of BCSCs in MCF-7,and promoted JMJD3 expression in BCSCs. However,GSK-J1 completely reflected the reaction. CONCLUSION JMJD3 can inhibit the proliferation of MCF-7,and the inhibition may be related to the reduced proportion of BCSCs.
作者
胡楠
邓力
包旭
廉冰
江育莹
张建华
汪宏
HU Nan;DENG Li;BAO Xu;LIAN Bing;JIANG Yuying;ZHANG Jianhua;WANG Hong(West China School of Pharmacy,Sichuan University,Chengdu,Sichuan,610041 P.R.China;State Key Laboratory of Biotherapy,Huaxi Hospital,Sichuan University,Chengdu,Sichuan,610041 P.R.China;Sichuan Center for Food and Drug Evaluation,Inspection ＆ Monitoring,Chengdu,Sichuan,610017 P.R.China)
出处
《华西药学杂志》
CAS
CSCD
2018年第5期485-489,共5页
West China Journal of Pharmaceutical Sciences
