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miRNA-18a 和 miRNA-4802 通过自噬抑制淋巴瘤细胞耐药性的

机制研究 Δ

1，2 #
周仕霞 1，2＊ ，李晓明 ，唐君玲 （1.西南医科大学附属医院血液内科，四川 泸州 646000；2.西南医科大学附属
1，2
医院干细胞实验室，四川 泸州 646000）

中图分类号 R966 文献标志码 A 文章编号 1001-0408（2022）11-1330-08
DOI 10.6039/j.issn.1001-0408.2022.11.09

摘 要 目的 研究miRNA-18a和miRNA-4802通过自噬对淋巴瘤细胞耐药性的调控机制。方法 以人Burkitt’s淋巴瘤细胞Daudi
和人套细胞淋巴瘤细胞JeKo-1为研究对象，阿霉素（ADR）、长春新碱（VCR）为实验药物。以ADR、VCR处理后，采用CCK-8法考
察上述2种细胞的相对活性，采用Western blot法检测凋亡标志蛋白活化型胱天蛋白酶9（cleaved caspase-9）和cleaved caspase-6的
表达，考察2种细胞对ADR、VCR的耐药性。采用自噬相关蛋白LC3-Ⅱ、p62表达水平检测、自噬流实验和透射电镜观察考察2种
细胞自噬活性的差异；采用荧光定量聚合酶链式反应考察2种细胞中miRNA-18a、miRNA-4802及unc-51样激酶（ULK1）、自噬相
关蛋白7（ATG7）mRNA的表达差异，并检测ULK1、ATG7蛋白表达差异。再以JeKo-1细胞为研究对象，考察经2种模拟生物体内
源的miRNAs（miRNA-18a mimics、miRNA-4802 mimics）处理后，细胞自噬活性和耐药性的变化情况。结果 以ADR和VCR处理
后，JeKo-1细胞比Daudi细胞具有更强的耐药性和自噬活性。JeKo-1细胞中miRNA-18a、miRNA-4802表达水平均显著低于Daudi
细胞，ULK1、ATG7 mRNA 和蛋白表达水平均显著高于 Daudi 细胞（P＜0.001）。以 miRNA-18a mimics 和 miRNA-4802 mimics 处
理JeKo-1细胞后，细胞的自噬活性和耐药性均显著降低。结论 miRNA-18a和miRNA-4802可分别通过降低自噬启动基因ULK1
和ATG7的表达，抑制淋巴瘤细胞的自噬活性，从而降低淋巴瘤细胞对ADR和VCR的耐药性。
关键词 淋巴瘤细胞；耐药性；miRNA-18a；miRNA-4802；自噬；机制

Mechanism study on miRNA-18a and miRNA-4802 suppressing drug resistance of lymphoma cells via
inhibiting autophagy
1，2
1，2
1，2
ZHOU Shixia ，LI Xiaoming ，TANG Junling （1. Dept. of Hematology，the Affiliated Hospital of Southwest
Medical University，Sichuan Luzhou 646000，China；2. Stem Cell Laboratory，the Affiliated Hospital of
Southwest Medical University，Sichuan Luzhou 646000，China）
ABSTRACT OBJECTIVE To study the regulation mechanism of miRNA-18a and miRNA-4802 on drug resistance of lymphoma
cells via autophagy. METHODS Using human burkitt’s lymphoma cell Daudi and human mantle cell lymphoma cell JeKo-1 as the
research objects，adriamycin（ADR）and vincristine（VCR）as experimental drugs. After treatment of ADR and VCR，relative cell
viability was detected with CCK-8 kit；the expression of apoptosis marker protein activated cleaved caspase-9 and cleaved
caspase-6 were detected by Western blot assay. The drug resistances of the two cells to ADR and VCR were investigated. The
difference of autophagy activity between the two kinds of cells by expression detection of autophagy related proteins LC3-Ⅱ and
p62，autophagy flow experiment and transmission electron microscope observation. Fluorescence quantitative polymerase chain
reaction was used to investigate the expression differences of miRNA-18a and miRNA-4802，ULK1 and ATG7 mRNA in the two
cells，and to detect the expression differences of ULK1 and ATG7 proteins. Taking JeKo-1 cells as the research object，the changes
of autophagy activity and drug resistance were investigated after treatment with endogenous miRNAs （miRNA-18a mimics，
miRNA-4802 mimics）of two simulated organisms. RESULTS After ADR and VCR treatment，compared with Daudi cells，JeKo-1
cells had stronger drug resistance and autophagy activity. The expression of miRNA-18a and miRNA-4802 in JeKo-1 cells were
significantly lower than Daudi cells，mRNA and protein expression of ULK1 and ATG7 were significantly higher than those of
Daudi cells （P＜0.001）. After treatment of miRNA-18a mimics and miRNA-4802 mimics，the autophagy activities and drug
resistances of JeKo-1 cells were decreased significantly. CONCLUSIONS miRNA-18a and miRNA-4802 can decrease drug
resistances of lymphoma cells to ADR and VCR by reducing
Δ 基 金 项 目 ：四 川 省 重 点 研 发（重 大 科 技 专 项）项 目（No.
the expression of autophagy-initiating genes ULK1 and ATG7，
2019YFS0301）；泸州市科技计划项目（No.2020-JYJ-50）
＊研究实习员，硕士。研究方向：血液肿瘤。E-mail：15111933606 and inhibiting the autophagy activity of lymphoma cells.
@163.com KEYWORDS lymphoma cells； drug resistance； miRNA-
# 通信作者：副教授，博士。研究方向：血液肿瘤。E-mail：Poly- 18a；miRNA-4802；autophagy；mechanism
cloneRES_TJL@163.com

·1330 · China Pharmacy 2022 Vol. 33 No. 11 中国药房 2022年第33卷第11期