微小RNA-145 影响乳腺癌细胞系MCF-7增殖和凋亡的机制

齐玉林

doi:10.16098/j.issn.0529-1356.2020.01.010

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解剖学报 ›› 2020, Vol. 51 ›› Issue (1) : 58-61. DOI: 10.16098/j.issn.0529-1356.2020.01.010

肿瘤生物学

微小RNA-145 影响乳腺癌细胞系MCF-7增殖和凋亡的机制

齐玉林¹ 韩慧芳^2* 张海新³

作者信息 +

Mechanism of microRAN-145 on proliferation and apoptosis of breast cancer cell line MCF-7#br#

QI Yu-lin¹ HAN Hui-fang^2*ZHANG Hai-xin³

Author information +

文章历史 +

摘要

目的探讨微小RNA-145（miR-145）参与乳腺癌MCF-7细胞增殖与凋亡的机制。方法体外培养永生化乳腺癌细胞系MCF-7细胞，并转染miR-145。分别应用Real-time PCR与Western blotting检测mRNA和蛋白水平。细胞计数试剂盒8（CCK-8）检测各组细胞增殖水平，流式细胞术检测不同处理组MCF-7细胞的凋亡水平。结果 Real-time PCR结果表明，高表达miR-145对MCF-7细胞Caspase-3、增殖细胞核抗原（PCNA）和Bcl-2的mRNA水平并无影响；而Western blotting实验结果表明，与对照组相比，转染miR-145 96 h可显著提高Caspase-3的表达，并抑制PCNA与Bcl-2的表达。CCK-8实验结果表明，过表达miR-145 72 h、96 h 后MCF-7细胞增殖率降低，差异具有统计学意义(P<0.05)。流式细胞术结果表明，过表达组MCF-7细胞的凋亡率显著高于对照组，差异具有统计学意义(P<0.05)。结论 miR-145可通过下调PCNA与Bcl-2蛋白、上调Caspase-3蛋白的表达，从而抑制MCF-7细胞增殖和促进MCF-7细胞凋亡，有望成为乳腺癌治疗的新靶点。

Abstract

Objective To explore the mechanism of microRNA-145 (miR-145) involved in proliferation and apoptosis of breast cancer MCF-7 cells. Methods The immortalized breast cancer cell line MCF-7 cells were cultured in vitro and transfected with miR-145. Real-time PCR and Western blotting were used to detect mRNA and protein levels, respectively. The proliferation level of each group was detected by cell counting kit-8 (CCK-8) method , and the apoptosis level of MCF-7 cells in different treatment groups was detected by flow cytometer. Results Real-time PCR result showed that miR-145 did not affect Caspase-3, proliferating cell nuclear antigen (PCNA) and B cell lymphoma factor 2 (Bcl-2) mRNA levels in MCF-7 cells. Western blotting analysis showed that compared with the control group, transfection of miR-145 for 96 hours significantly increased the expression of Caspase-3 and inhibited the expression of PCNA and Bcl-2. The result of CCK-8 assay showed that the proliferation rate of MCF-7 cells was decreased after overexpression of miR-145 for 72 hours and 96 hours (P<0.05). The result of flow cytometer showed that the apoptosis rate of MCF-7 cells in overexpression group was significantly higher than that in the control group (P<0.05). Conclusion MiR-145 can inhibit the proliferation and promote the apoptosis of MCF-7 cells by down-regulating PCNA and Bcl-2 and up-regulating the expression of Caspase-3, which may be a new target for breast cancer treatment.

导出引用

齐玉林韩慧芳张海新. 微小RNA-145 影响乳腺癌细胞系MCF-7增殖和凋亡的机制[J]. 解剖学报. 2020, 51(1): 58-61 https://doi.org/10.16098/j.issn.0529-1356.2020.01.010

QI Yu-lin HAN Hui-fang ZHANG Hai-xin. Mechanism of microRAN-145 on proliferation and apoptosis of breast cancer cell line MCF-7#br#[J]. Acta Anatomica Sinica. 2020, 51(1): 58-61 https://doi.org/10.16098/j.issn.0529-1356.2020.01.010

中图分类号： R736.8

参考文献

［1］ Chen WQ, Zheng RSh. Incidence,mortality and survival analysis of breast cancer in China［J］. Chinese Journal of Clinical Oncology, 2015,42(13):668-674.(in Chinese)

陈万青,郑荣寿.中国女性乳腺癌发病死亡和生存状况［J］.中国肿瘤临床,2015,42(13):668-674.

［2］ Bray F, Ferlay J, Soerjomataram Ⅰ, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries ［J］. CA Cancer J Clin, 2018,68(6):394-424.

［3］ Li RP, Zeng Zh, Zhao M, et al. Expression and clinical significance of Yes-related protein 1 in breast cancer［J］. Anatomy Research,2016,38(6):468-470, 474, 512. (in Chinese)

李瑞平,曾铮,赵敏, 等.Yes相关蛋白1在乳腺癌中的表达及临床意义［J］.解剖学研究,2016,38(6):468-470, 474, 512.

［4］ Huang YY, Bao Y. Experience breast cancer: from “disease“to “disability” of the female body［J］. Society, 2013,33(2):185-207. (in Chinese)

黄盈盈,鲍雨.经历乳腺癌:从“疾病”到“残缺”的女性身体［J］.社会,2013,33(2):185-207.

［5］ Zhang Q, Wu ZhSh, Zhang GH, et al. Expression of miR-145 in breast carcinoma and its significance［J］. Chinese Journal of Clinical and Experimental Pathology, 2009,25(1):9-12. (in Chinese)

张晴,吴正升,张瑰红,等.miR-145在乳腺癌中表达及意义［J］.临床与实验病理学杂志,2009,25(1):9-12.

［6］ Luo QC, Zhang H, Chen RSh, et al. Mechanism and function of microRNA mir-145 regulating Nanog in breast cancer cells［J］. Journal of Changsha University,2016,30(5):10-13, 16. (in Chinese)

骆启聪,张环,陈蓉珊,等.微小RNA mir-145在乳腺癌细胞中调控Nanog的机制与功能研究［J］.长沙大学学报,2016,30(5):10-13, 16.

［7］ Zhang BJ, Jin YX, Ma ZhL. The role of microRNA in tumors［J］. Chinese Journal of Nature,2014,36(5):364-372. (in Chinese)

张冰洁,金由辛,马中良.microRNA在肿瘤中的作用［J］.自然杂志,2014,36(5):364-372.

［8］ Ma ShY, Bai Y, Han N, et al. Recent research progress of biogenesis and functions of miRNA *［J］. Hereditas,2012,34(4):5-10. (in Chinese)

马圣运,白玉,韩凝,等.miRNA *生物合成及其功能研究的新发现［J］.遗传,2012,34(4):5-10.

［9］ Wang HCh, Zhang HJ, Hu ChJ. Roles of miRNA in breast cancer metastasis［J］. Chemistry of Life,2012,32(3):205-210. (in Chinese)

王焕春,张海静,胡成进.MiRNA在乳腺癌转移中的作用［J］.生命的化学,2012,32(3):205-210.

［10］Chen B, Wei WD. Advances in the role of miRNAs in breast cancer drug resistance and related mechanisms［J］. Guangdong Medical Journal,2014,35(17):2791-2794. (in Chinese)

陈博,韦尉东.miRNA在乳腺癌耐药中的作用及其相关机制的研究进展［J］.广东医学,2014,35(17):2791-2794.

［11］Zhang J. Functional study of miRNAs involved in breast cancer cell proliferation and metastasis［D］. Fudan University,2008. (in Chinese)

张进. 乳腺癌细胞增殖与转移相关miRNA的功能研究［D］.复旦大学,2008.

［12］Zhao XA, Hu JH, Zhao XH. Effect of Hsa-miR-145 on invasion and migration potential in triple-negative breast cancer cell［J］. Chinese Journal of Clinicians(Electronic Edition),2012,6(15):4276-4279. (in Chinese)

赵晓艾,胡金华,赵新汉.Hsa-miR-145对人三阴性乳腺癌细胞侵袭和迁移的影响［J］.中华临床医师杂志(电子版),2012,6(15):4276-4279.

［13］Ding J. Targeted regulation of BCL2L2 in breast cancer by miR-145［D］. East China Normal University,2016. (in Chinese)

丁健. miR-145在乳腺癌中对BCL2L2的靶向调控［D］.华东师范大学,2016.

［14］Feng W, Wang C, Liang C, et al. The dysregulated expression of KCNQ1OT1 and its interaction with downstream factors miR-145/CCNE2 in breast cancer cells ［J］. Cell Physiol Biochem, 2018,49(2):432-446.

［15］Quan Y, Huang X, Quan X. Expression of miRNA-206 and miRNA-145 in breast cancer and correlation with prognosis ［J］. Oncol Lett, 2018,16(5):6638-6642.

［16］Yamada Y, Koshizuka K, Hanazawa T, et al. Passenger strand of miR-145-3p acts as a tumor-suppressor by targeting MYO1B in head and neck squamous cell carcinoma ［J］. Int J Oncol, 2018,52(1):166-178.