[1]雷优扬,周华,代朦,等.miR-382通过PGC-1α对三阴性乳腺癌细胞株4T1生物学特性的影响[J].第三军医大学学报,2019,41(15):1405-1422.
 LEI Youyang,ZHOU Hua,DAI Meng,et al.MiR-382 inhibits proliferation and migration of triple negative breast cancer cell line 4T1 by mediating PGC-1α[J].J Third Mil Med Univ,2019,41(15):1405-1422.
点击复制

miR-382通过PGC-1α对三阴性乳腺癌细胞株4T1生物学特性的影响(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第15期
页码:
1405-1422
栏目:
基础医学
出版日期:
2019-08-15

文章信息/Info

Title:
MiR-382 inhibits proliferation and migration of triple negative breast cancer cell line 4T1 by mediating PGC-1α
作者:
雷优扬周华代朦靳鑫明佳
重庆医科大学附属第二医院乳腺甲状腺外科
Author(s):
LEI Youyang ZHOU Hua DAI Meng JIN Xin MING Jia

Department of Mammary Gland and Thyroid, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China

关键词:
三阴性乳腺癌microRNA-382PGC-1&alpha
Keywords:
triple negative breast cancer microRNA-382 PGC-1&alpha
分类号:
R394.3; R730.23; R737.9
文献标志码:
A
摘要:
 
目的 研究乳腺癌4T1细胞内miR-382下调PGC-1α对细胞生物学特性的影响。方法 ①以4T1细胞作为研究对象,miR-382mimics或抑制性探针anti-miR-382转染4T1细胞后,以qRT-PCR检测4T1中miR-382水平变化,qRT-PCR和Western blot检测各组PGC-1α的表达水平变化;②荧光素酶报告基因检测miR-382与PGC-1α 3′UTR的结合位点;③将miR-382mimics或对照miR分别与pCDNA3.1或pCDNA-PGC1α共转染于4T1细胞,Transwell迁移实验检测细胞迁移能力的变化;CCK8 实验检测细胞增殖活力的变化;皮下移植瘤实验和尾静脉肺移植瘤实验分别检测小鼠体内肿瘤生长及肺转移的变化。结果 转染miR-382 mimics 之后乳腺癌细胞PGC-1α表达明显降低(P<0.05),转染抑制性探针anti-miR-382后PGC-1α表达明显升高(P<0.05);miR-382与PGC-1α存在靶向调控核心序列,其作用位点位于PGC-1α 3′UTR 565-582(核心序列ACAACTT)。相比对照+pCDNA3.1组,miR-382+pCDNA3.1组的细胞迁移和增殖活力明显降低,4T1皮下移植瘤体积和尾静脉移植瘤肺转移灶的面积显著减小(P<0.05);对照+pCDNA-PGC-1α较对照+pCDNA3.1组,细胞迁移和增殖活力明显提高,4T1皮下移植瘤体积和尾静脉移植瘤肺转移灶的面积显著增加(P<0.05);miR-382的抑瘤效应因PGC-1α过表达而被阻断。结论 miR-382可能通过PGC-1α抑制4T1细胞株的增殖和肺转移能力,其作用位点可能位于PGC-1α 3′UTR 565-582。
Abstract:

Objective To explore the effect of miR-382 down-regulating PGC-1α on the biological behaviors of triple negative breast cancer 4T1 cells. Methods① After miR-382 mimics or inhibitory probe anti-miR-382 was transfected into 4T1 cells, the expression level of miR-382 was detected by qRT-PCR. The changes of PGC-1α expression at mRNA and protein levels were detected by qRT-PCR and Western blotting, respectively. ② Luciferase reporter gene assay was adopted to identify the binding site of miR-382 mimics to PGC-1α 3′UTR. ③miR-382 mimics or control miR were co-transfected into 4T1 cells with pCDNA3.1 or pCDNA-PGC1α, respectively. Transwell migration assay, CCK8 assay and subcutaneous xenografts and tail vein lung xenografts were respectively used to detect the changes in cell invasiveness, cell proliferation, and tumor growth and lung metastasis in mice. ResultsThe expression of PGC-1α was significantly decreased in 4T1 cells after transfection with miR-382 mimics (P<0.05), and it was obviously increased after transfection with anti-miR-382 (P<0.05). There was a targeted regulatory core sequence (ACAACTT) of miR-382 and PGC-1α, and the site was at PGC-1α 3′UTR 565-582. Compared with the control +pCDNA3.1 group, the cell migration and proliferation activity of the miR-382+pCDNA3.1 group was significantly decreased, and the size of the 4T1 subcutaneous tumor and the area of metastatic tumor in the lung were notably decreased (P<0.05). Compared with the control +pCDNA3.1 group, the cell migration and proliferation activity were significantly increased in the miR-382+pCDNA-PGC-1α group, and the size of 4T1 subcutaneous tumor and the area of metastatic tumor in the lung was significantly increased (P<0.05); The antitumor effect of miR-382 was blocked by overexpression of PGC-1α. ConclusionMiR-382 inhibits the proliferation and lung metastasis of 4T1 cells by down-regulating PGC-1α, and the interaction site may be PGC-1α 3′UTR 565-582.

参考文献/References:

[1]BOSCH A, EROLES P, ZARAGOZA R, et al. Triple-negative breast cancer: molecular features, pathogenesis, treatment and current lines of research.[J]. Cancer Treatment Rev, 2010, 36(3):206-215.
[2]LEBLEU V S, O’CONNELL J T, GONZALEZ HERRERA K N,et al. PGC-1α mediates mitochondrial biogenesis and oxidative phosphorylation in cancer cells to promote metastasis[J]. Nat Cell Biol,2014,16( 10) : 992-1003. DOI: 10.1038/ncb3039.
[3]PUIGSERVER P,WU Z,PARK C W,et al. A cold-inducible coactivator of nuclear receptors linked to adaptive thermogenesis[J]. Cell,1998,92(6): 829-839. DOI: 10.1016/S0092-8674(00)81410-5.
[4]CALIN G A,CROCE C M. MicroRNA signatures in human cancers[J]. Nat Rev Cancer,2006,6(11): 857-866. DOI:10.1038/nrc1997.
[5]FU L, LI Z, ZHU J, et al. Serum expression levels of microRNA-382-3p, -598-3p, -1246 and -184 in breast cancer patients.[J]. Oncol Letters, 2016, 12(1):269. DOI: 10.3892/ol.2016.4582. 
[6]代朦,靳鑫,雷优扬,等.过表达miR-382的肿瘤相关巨噬细胞对三阴性乳腺癌生物学特性的影响[J].第三军医大学学报,2018,40(15):1375-1382. DOI:10.16016/j.1000-5404. 201801107.
DAI M,JIN X,LEI Y Y, et al. Effect of miR-382 overexpressing tumor-associated macrophages on biological properties of triple-negative breast cancer 4T1 cells [J].J Third Mil Med Univ,2018,40(15):1375-1382. DOI:10.16016/j.1000-5404. 201801107.
[7]靳鑫,倪田根,王宁,等.PGC-1α通过线粒体介导巨噬细胞极化状态的机制研究[J].第三军医大学学报,2019,41(1):56-62. DOI:10.16016/j.1000-5404. 201808139.
JIN X, NI T G,WANG N,et al. Mechanism of mitochondria-mediated PGC-1α in macrophage polarization[J].J Third Mil Med Univ,2019,41(01):56-62.DOI:10.16016/j.1000-5404. 201808139.
[8]FAN L, STRASSER-WEIPPL K, LI J J, et al. Breast cancer in China[J]. Lancet Oncology, 2014, 15(7):e279-e289. DOI:10.1016/S1470-2045(13) 70567-9.
[9]COSTA R,SHAH A N,SANTA-MARIA C A,et al.  Targeting Epidermal Growth Factor Receptor in triple negative breast cancer: New discoveries and practical insights for drug development[J]. Cancer Treatment Rev, 2017, 53:111-119. DOI:10.1016/j.ctrv.2016.12.010.
[10]DENKERT C,LIEDTKE C,TUTT A,et al. Molecular alterations in triple-negative breast cancer-the road to new treatment strategies[J]. Lancet,2017,389(10087): 2430-2442. DOI:10.1016/S0140-6736(16)32454-0.
[11]LEBLEU V S,O’CONNELL J T,GONZALEZ HERRERA K N,et al. PGC-1α mediates mitochondrial biogenesis and oxidative phosphorylation in cancer cells to promote metastasis[J]. Nat Cell Biol,2014,16(10) : 992-1003. DOI:10.1038/ncb3039.
[12]PUIGSERVER P, WU Z, PARK C W, et al. A cold-inducible coactivator of nuclear receptors linked to adaptive thermogenesis[J]. Cell, 1998, 92(6):829-839. DOI:10.1016/S0092-8674(00)81410-5.
[13]ST-PIERRE J, DRORI S, ULDRY M,et al. Suppression of reactive oxygen species and neurodegeneration by the PGC-1α transcriptional coactivators[J]. Cell,2006,127(2):397-408. DOI:10.1016/j.cell.2006.09.024.
[14]YU Z, BASERGA R, CHEN L, et al. MicroRNA, cell cycle, and human breast cancer[J]. Am J Pathol, 2010,176(3):1058-1064.DOI: 10.2353/ajpath.2010.090664.
[15]STENVANG J, PETRI A, LINDOW M, et al. Inhibition of microRNA function by antimiR oligonucleotides[J]. Silence, 2012, 3(1):1. DOI: 10.1186/1758-907X-3-1.
[16]ZHANG H, LI Y, LAI M. The microRNA network and tumor metastasis. Oncogene, 2010,29:937-948. DOI: 10.1038/onc.2009.406. 
[17]JOVANOVIC M, HENGARTNER M O. miRNAs and apoptosis: RNAs to die for. Oncogene, 2006,25:6176-6187. DOI: 10.1038/sj.onc.1209912.
[18]LI Y, HONG F, YU Z. Decreased expression of microRNA-206 in breast cancer and its association with disease characteristics and patient survival[J]. J Int Med Res, 2013, 41(3):596-602. DOI: 10.1177/0300060513485856.
[19]KONG Y W, FERLAND-MCCOLLOUGH D, JACKSON T J, et al. MicroRNAs in cancer management[J]. Lancet Oncol. 2012, 13(6):e249-e258. DOI: 10.1016/S1470-2045(12) 70073-6.
[20]CHEN T, REN H, THAKUR A, et al. MiR-382 inhibits tumor progression by targeting SETD8 in non-small cell lung cancer[J]. Biomedicine&Pharmacotherapy, 2017, 86:248-253. DOI:10.1016/j.biopha.2016.12.007. 
[21]ZHOU B, SONG J, HAN T, et al. MiR-382 inhibits cell growth and invasion by targeting NR2F2 in colorectal cancer[J]. Mol Carcinogen, 2016,55(12) : 2260- 2267. DOI:10.1002/mc.22466.
 

相似文献/References:

[1]代朦,靳鑫,雷优扬,等.过表达miR-382的肿瘤相关巨噬细胞对三阴性乳腺癌生物学特性的影响[J].第三军医大学学报,2018,40(15):1375.
 DAI Meng,JIN Xin,LEI Youyang,et al.Effect of miR-382 overexpressing tumor-associated macrophages on biological properties of triple-negative breast cancer 4T1 cells [J].J Third Mil Med Univ,2018,40(15):1375.

更新日期/Last Update: 2019-08-13