miR-223-3p通过调控ECT2诱导非小细胞肺癌细胞凋亡的实验研究

doi:10.3969/j.issn.1004-616x.2021.03.005

癌变·畸变·突变 ›› 2021, Vol. 33 ›› Issue (3): 187-192.doi: 10.3969/j.issn.1004-616x.2021.03.005

miR-223-3p通过调控ECT2诱导非小细胞肺癌细胞凋亡的实验研究

王熙凯¹, 孟庆贺², 高燕鲁³

1. 首都医科大学基础医学院, 北京 100069;
2. 北京大学公共卫生学院, 北京 100191;
3. 山东中医药大学第二附属医院, 山东济南 250001

收稿日期:2021-01-21 修回日期:2021-04-27 出版日期:2021-05-30 发布日期:2021-06-09
通讯作者: 高燕鲁，E-mail：shdaiw@sina.com E-mail:shdaiw@sina.com
作者简介:王熙凯，E-mail：905301452@qq.com。

miR-223-3p negatively regulated ECT2 to induce apoptosis in non-small cell lung cancer cells

WANG Xikai¹, MENG Qinghe², GAO Yanlu³

1. School of Basic Medical Science, Capital Medical University, Beijing 100069;
2. School of Public Health, Peking University, Beijing 100191;
3. The Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan 250001, Shandong, China

Received:2021-01-21 Revised:2021-04-27 Online:2021-05-30 Published:2021-06-09

摘要/Abstract

摘要： 目的：探讨miR-223-3p对非小细胞肺癌细胞增殖与凋亡的影响及其可能的作用机制。方法：选取24例非小细胞肺癌患者的癌及癌旁组织，实时定量PCR检测miR-223-3p的相对表达水平，免疫组化检测分析上皮细胞转化序列2（ECT2）蛋白的表达水平，并对两者进行相关性分析。将人非小细胞肺癌A549细胞分为正常对照组、转染对照组、miR-223-3p过表达组，其中正常对照组细胞未作任何处理，转染对照组细胞转染空质粒载体，miR-223-3p过表达组转染miR-223-3p mimics。MTT实验和平板集落形成实验检测各组细胞的增殖率，流式细胞术检测各组细胞的凋亡率，Western blot实验检测ECT2蛋白的表达水平；采用siRNA转染法沉默A549细胞中的ECT2后，检测细胞增殖及凋亡的变化。结果：miR-223-3p在癌组织中表达显著低于癌旁组织，而癌组织中ECT2蛋白表达高于癌旁组织，二者存在负相关关系（r=-0.666，P < 0.05）；与正常对照组和转染对照组细胞相比，miR-223-3p过表达组细胞的增殖率降低（P < 0.05），ECT2蛋白表达显著降低，而细胞凋亡率显著升高（P < 0.05）；沉默ECT2对细胞增殖和凋亡的影响与过表达miR-223-3p基本一致。结论：miR-223-3p可能通过负向调控ECT2的表达，进而抑制非小细胞肺癌细胞增殖并诱导其凋亡，其作用机制有待进一步研究。

关键词: miR-223-3p, 上皮细胞转化序列2, 非小细胞肺癌, 细胞凋亡

Abstract: OBJECTIVE： To investigate effects of miR- 223- 3p on proliferation and apoptosis of nonsmall cell lung cancer cells. METHODS： Relative expression levels of miR-223-3p were detected by realtime quantitative PCR in 24 patients with non-small cell lung cancer (NSCLC) and expression levels of epithelial transformation sequence 2 (ECT2) protein were analyzed by immunohistochemistry. Correlations between the expression levels of miR-223-3p and ECT2 were analyzed. Human NSCLC cells A549 were divided into normal control group， transfected control group and miR-223-3p over-expression group. The normal control group cells were not treated and cultured normally，and the transfected control group cells were transfected with empty plasmid vector using transfection reagents. The miR-223-3p over-expression group was transfected with miR-223-3p mimics. Proliferation rates of cells from each group were detected by the MTT and the plate colony formation assays. Apoptosis rates of cells in each group were detected by flow cytometry. Expression levels of the ECT2 protein were detected by Western blot assay. After ECT2 was knocked down by siRNA， A549 cells were divided into normal control， transfected control and ECT2 silenced groups. Then， changes of cell proliferation and apoptosis in each group were detected. RESULTS： Expressions of miR-223- 3p in cancer tissues were significantly lower than that in paracancerous tissues while expressions of ECT2 protein were higher， showing a negative correlation between them (r=- 0.666， P < 0.05). Compared with the normal control and the transfected control groups， proliferation rates of miR- 223- 3p in the over- expression group was decreased (P < 0.05)，expressions of ECT2 protein were significantly decreased，and apoptosis rates were significantly increased (P < 0.05). Effects from siRNA knockdown of ECT2 on cell proliferation and apoptosis were basically the same as that of miR- 223-3p over-expression group. CONCLUSION： Our data show that miR- 223-3p inhibited proliferation and induced apoptosis of NSCLC cells by negatively regulating the expression of ECT2.

Key words: miR-223-3p, ECT2, non-small cell lung cancer, apoptosis

中图分类号:

R734.2

王熙凯, 孟庆贺, 高燕鲁. miR-223-3p通过调控ECT2诱导非小细胞肺癌细胞凋亡的实验研究[J]. 癌变·畸变·突变, 2021, 33(3): 187-192.

WANG Xikai, MENG Qinghe, GAO Yanlu. miR-223-3p negatively regulated ECT2 to induce apoptosis in non-small cell lung cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(3): 187-192.

参考文献

[1] SUNG H, FERLAY J, SIEGEL R L, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countiries[J]. CA Cancer J Clin, 2021, 71(3): 209-249.
[2] RECK M, RABE K F. Precision diagnosis and treatment for advanced non-small-cell lung cancer[J]. N Engl J Med, 2017, 377(9): 849-861.
[3] GAMERITH G, KOCHER F, RUDZKI J, et al. ASCO 2018 NSCLC highlights-combination therapy is key[J]. Memo, 2018, 11(4): 266-271.
[4] LI Y L, WANG D T, LI X, et al. MiR-199a-5p suppresses nonsmall cell lung cancer via targeting MAP3K11[J]. J Cancer, 2019, 10(11): 2472-2479.
[5] 戚欣, 王会子, 陈旭东, 等. miR-223-3p通过靶向RAC1调控肝细胞癌SMMC-7721细胞的增殖和凋亡[J]. 中国肿瘤生物治疗杂志, 2020, 27(6): 664-670.
[6] ZHAO Y L, FANG X, FANG H, et al. ATPR-induced G0/G1 phase arrest in gastric cancer cells by regulating the binding of 14-3-3ε and filamin A[J]. Cancer Med, 2018, 7(7): 3373-3384.
[7] 沈存思, 杨福州, 项莹颖, 等. 中药活性化合物高良姜素与吉非替尼抗非小细胞肺癌的协同增效作用及机制研究[J]. 南京中医药大学学报, 2021, 37(1): 72-76.
[8] WU K L, TSAI Y M, LIEN C T, et al. The roles of microRNA in lung cancer[J]. Int J Mol Sci, 2019, 20(7): E1611.
[9] PINHO J D, SILVA G E B, TEIXEIRA JÚNIOR A A L, et al. MIR-107, MIR-223-3P and MIR-21-5P reveals potential biomarkers in penile cancer[J]. Asian Pac J Cancer Prev, 2020, 21(2): 391-397.
[10] LI S, FENG Y, HUANG Y, et al. miR-223-3p regulates cell viability, migration, invasion, and apoptosis of non-small cell lung cancer cells by targeting RHOB[J]. Open Life Sci, 2020, 15(1): 389-399.
[11] TATSUMOTO T, XIE X, BLUMENTHAL R, et al. Human ECT2 is an exchange factor for Rho GTPases, phosphorylated in G2/M phases, and involved in cytokinesis[J]. J Cell Biol, 1999, 147(5): 921-928.
[12] KOSIBATY Z, MURATA Y, MINAMI Y, et al. ECT2 promotes lung adenocarcinoma progression through extracellular matrix dynamics and focal adhesion signaling[J]. Cancer Sci, 2021, 112(2): 703-714.
[13] LI Q P, JIANG B, QI Y, et al. Long non-coding RNA SLCO4A1-AS1 drives the progression of non-small-cell lung cancer by modulating miR-223-3p/IKKα/NF-κB signaling[J]. Cancer Biol Ther, 2020, 21(9): 806-814.
[14] JUSTILIEN V, LEWIS K C, MENESES K M, et al. Protein kinase C_ι promotes UBF₁-ECT2 binding on ribosomal DNA to drive rRNA synthesis and transformed growth of non-smallcell lung cancer cells[J]. J Biol Chem, 2020, 295(24): 8214-8226.
[15] 李伦, 兴成娟, 丛玲, 等. miR-223-3p靶向上皮细胞转化序列2基因调控胃癌细胞周期和凋亡的相关性研究[J]. 世界华人消化杂志, 2018, 26(2): 71-79.
[16] SUN Q Y, DING L W, JOHNSON K, et al. SOX7 regulates MAPK/ERK-BIM mediated apoptosis in cancer cells[J]. Oncogene, 2019, 38(34): 6196-6210.
[17] 黄尚校, 李春君, 黄剑锋, 等. miR-1-3p通过靶向调控CDK9对非小细胞肺癌细胞增殖和凋亡的影响[J]. 中国免疫学杂志, 2020, 36(20): 2468-2472, 2478.

miR-223-3p通过调控ECT2诱导非小细胞肺癌细胞凋亡的实验研究

miR-223-3p negatively regulated ECT2 to induce apoptosis in non-small cell lung cancer cells

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