诊断学理论与实践

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2018 , Vol. 17 >Issue 01: 70 - 75

DOI: https://doi.org/10.16150/j.1671-2870.2018.01.013

论著

肿瘤高甲基化基因1重要下游基因HMMR对乳腺癌细胞株MDA-MB-231迁移和侵袭能力影响的体外实验

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  • 上海交通大学医学院附属第九人民医院外科,上海 200001

收稿日期: 2017-12-25

  网络出版日期: 2018-02-25

基金资助

国家自然科学基金（81572760）; 上海市科委自然科学基金（14ZR1424400）

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Effect of hypermethylated in cancer 1-downstream-regulated gene HMMR on cell migration and invasion in breast cancer: an in vitro study on breast cancer cell line MDA-MB-231

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  • Department of Surgery, The Ninth People's Hospital of Shanghai Jiao Tong University School of Medicine, Shanghai 200001, China

Received date: 2017-12-25

  Online published: 2018-02-25

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摘要

目的：探讨肿瘤高甲基化基因1（hypermethylated in cancer 1,HIC-1）与透明质酸介导的细胞游走受体（hyaluronan-mediated motility receptor,HMMR）基因之间的调控关系,观察HMMR产物对MDA-MB-231乳腺癌细胞株的迁移和侵袭能力的影响。方法：MDA-MB-231细胞分别转染双链小激活RNA（small activating RNA,saRNA） HIC-1和双链小干扰RNA（small interfering RNA,siRNA） HMMR,通过反转录聚合酶链反应（reverse transcription polymerase chain reaction,RT-PCR）和蛋白印迹法检测观察转染后HIC-1、HMMR的mRNA和蛋白的表达情况,用CCK-8细胞增殖实验检测HIC-1和HMMR表达改变对乳腺癌细胞增殖能力的影响,再用Transwell小室观察其改变对乳腺癌细胞迁移和侵袭能力的影响。结果：转染saRNA HIC-1时MDA-MB-231细胞中HIC-1上调,同时HMMR表达下调,细胞的迁移和侵袭能力受抑制,细胞增殖能力未发生改变;转染siRNA HMMR时,MDA-MB-231细胞中HMMR表达下调,细胞的迁移和侵袭能力受抑制,细胞增殖能力未受影响。结论：HIC-1可调控HMMR的表达,HIC-1表达上调可下调HMMR,乳腺癌细胞的迁移和侵袭能力受抑制;乳腺癌细胞中因HIC-1失活,导致HMMR呈高表达,促进乳腺癌细胞的迁移和侵袭。

关键词： 乳腺癌; 高甲基化基因1; 透明质酸介导的细胞游走受体; MDA-MB-231细胞株

本文引用格式

裴文江, 赵峰, 吴衍, 钟明, 高航, 李幼生, 顾岩, 郭善禹, 戴谦诚, 张伟 . 肿瘤高甲基化基因1重要下游基因HMMR对乳腺癌细胞株MDA-MB-231迁移和侵袭能力影响的体外实验[J]. 诊断学理论与实践, 2018 , 17(01) : 70 -75 . DOI: 10.16150/j.1671-2870.2018.01.013

Abstract

Objective: To investigate the regulatory relationship between hypermethylated in cancer 1 (HIC-1) and hyaluronan-mediated motility receptor (HMMR) and to study the effect of HMMR on migration and invasion of MDA-MB-231 breast cancer cell strains. Methods: MDA-MB-231 cells were transfected with double-strand saRNA HIC-1 and double-strand siRNA HMMR, respectively. The expression of HIC-1 and HMMR in MDA-MB-231 cells were detected by RT-PCR and Western blotting. CCK-8 assay was used to detect cell proliferation after siRNA or saRNA transfection. Transwell assay was used to determine the effect of siRNA or saRNA transfection on migration and invasion of MDA-MB-231 breast cancer cells. Results: MDA-MB-231 breast cancer cells were successfully transfected. The up-regulation of HIC-1 induced HMMR down-regulation, and significantly inhibited cell migration and invasion of MDA-MB-231 breast cancer cells. HMMR was down-regulated by siRNA HMMR transfection leading to inhibition of cell migration and invasion. However, cell proliferation have not been affected in both saRNA HIC-1 and saRNA HMMR transfections. Conclusions: HMMR is regulated by HIC-1 and it plays an important role in cell migration and invasion. Inhibition of HIC-1 upregulates HMMR, thus promotes cell migration and invasion of breast cancer cells.

Key words： Breast cancer; Hypermethylated in cancer 1; Hyaluronan-mediated motility receptor; MDA-MB-231 cell strains

参考文献

[1] Harbeck N, Gnant M.Breast cancer[J]. Lancet,2017,389(10074):1134-1150.
[2] Ferlay J, Soerjomataram I, Dikshit R, et al.Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012[J]. Int J Cancer,2015,136(5):E359-E386.
[3] Fan L, Strasser-Weippl K, Li JJ, et al.Breast cancer in China[J]. Lancet Oncol,2014,15(7):e279-e289.
[4] Chen W, Zheng R, Baade PD, et al.Cancer statistics in China, 2015[J]. CA Cancer J Clin,2016,66(2):115-132.
[5] Parrella P, Scintu M, Prencipe M, et al. HIC1 promoter methylation and 17p13.3 allelic loss in invasive ductal carcinoma of the breast[J]. Cancer Lett,2005,222(1):75-81.
[6] van Rechem C, Boulay G, Pinte S, et al. Differential regu-lation of HIC1 target genes by CtBP and NuRD, via an acetylation/SUMOylation switch, in quiescent versus proliferating cells[J]. Mol Cell Biol,2010,30(16):4045-4059.
[7] Li P, Liu X, Dong ZM, et al.Epigenetic silencing of HIC1 promotes epithelial-mesenchymal transition and drives progression in esophageal squamous cell carcinoma[J]. Oncotarget,2015,6(35):38151-38165.
[8] Stevens LE, Cheung WKC, Adua SJ, et al.Extracellular Matrix Receptor Expression in Subtypes of Lung Adenocarcinoma Potentiates Outgrowth of Micrometastases[J]. Cancer Res,2017,77(8):1905-1917.
[9] Thangavel C, Boopathi E, Liu Y, et al.RB Loss Promotes Prostate Cancer Metastasis[J]. Cancer Res,2017,77(4):982-995.
[10] Hardwick C, Hoare K, Owens R, et al.Molecular cloning of a novel hyaluronan receptor that mediates tumor cell motility[J]. J Cell Biol,1992,117(6):1343-1350.
[11] Adamia S, Pilarski PM, Belch AR, et al.Aberrant spli-cing, hyaluronan synthases and intracellular hyaluronan as drivers of oncogenesis and potential drug targets[J]. Curr Cancer Drug Targets,2013,13(4):347-361.
[12] Zhao F, Pan S, Gu Y, et al.Small activating RNA restores the activity of the tumor suppressor HIC-1 on breast cancer[J]. PLoS One,2014,9(1):e86486.
[13] Kumar S.P53 induction accompanying G2/M arrest upon knockdown of tumor suppressor HIC1 in U87MG glioma cellsc[J]. Mol Cell Biochem,2014,395(1-2):281-290.
[14] Chen WY, Wang DH, Yen RC, et al.Tumor suppressor HIC1 directly regulates SIRT1 to modulate p53-dependent DNA-damage responses[J]. Cell,2005,123(3):437-448.
[15] Briggs KJ, Eberhart CG, Watkins DN.Just say no to ATOH: how HIC1 methylation might predispose medulloblastoma to lineage addiction[J]. Cancer Res,2008,68(21):8654-8656.
[16] Mohammad HP, Zhang W, Prevas HS, et al.Loss of a single Hic1 allele accelerates polyp formation in Apc(Δ716) mice[J]. Oncogene,2011,30(23):2659-2669.
[17] Vilgelm A, Wei JX, Piazuelo MB, et al.DeltaNp73alpha regulates MDR1 expression by inhibiting p53 function[J]. Oncogene,2008,27(15):2170-2176.
[18] van Rechem C, Rood B R, Touka M, et al. Scavenger chemokine(CXC motif) receptor 7(CXCR7) is a direct target gene of HIC1(hypermethylated in cancer 1)[J]. J Biol Chem,2009,284(31):20927-20935.
[19] Zeng S, Yang Y, Cheng X, et al.HIC1 epigenetically represses CIITA transcription in B lymphocytes[J]. Biochim Biophys Acta,2016,1859(12):1481-1489.
[20] Cheng G, He J, Zhang L, et al.HIC1 modulates uveal melanoma progression by activating lncRNA-numb[J]. Tumour Biol,2016,37(9):12779-12789.
[21] Assmann V, Jenkinson D, Marshall JF, et al.The intracellular hyaluronan receptor RHAMM/IHABP interacts with microtubules and actin filaments[J]. J Cell Sci,1999, 112( Pt 22):3943-3954.
[22] Zlobec I, Terracciano L, Tornillo L, et al.Role of RHAMM within the hierarchy of well-established prognostic factors in colorectal cancer[J]. Gut,2008,57(10):1413-1419.
[23] Tolg C, McCarthy JB, Yazdani A, et al. Hyaluronan and RHAMM in wound repair and the "cancerization" of stromal tissues[J]. Biomed Res Int,2014,2014:103923.
[24] Hall CL, Yang B, Yang X, et al.Overexpression of the hyaluronan receptor RHAMM is transforming and is also required for H-ras transformation[J]. Cell,1995,82(1):19-26.
[25] Chu TLH, Connell M, Zhou L, et al.Cell Cycle-Dependent Tumor Engraftment and Migration Are Enabled by Aurora-A[J]. Mol Cancer Res,2018,16(1):16-31.
[26] Entwistle KM, Silyn-Roberts H, Ochieng Abuodha S.The relative fracture strengths of the inner and outer surfaces of the eggshell of the domestic fowl[J]. Proc Biol Sci,1995,262(1364):169-174.
[27] Hao M, Li Y, Wang J, et al.HIC1 loss promotes prostate cancer metastasis by triggering epithelial-mesenchymal transition[J]. J Pathol,2017,242(4):409-420.
[28] Hu B, Zhang K, Li S, et al.HIC1 attenuates invasion and metastasis by inhibiting the IL-6/STAT3 signalling pathway in human pancreatic cancer[J]. Cancer Lett,2016,376(2):387-398.
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