沉默UHRF1对乳腺癌细胞增殖和转移的影响

doi:10.11904/j.issn.1002-3070.2017.06.006

摘要/Abstract

摘要： 目的探讨UHRF1对乳腺癌MDA-MB-231细胞增殖以及侵袭的影响及其相关机制。方法采用四甲基偶氮唑盐微量酶反应比色法(MTT)检测沉默UHRF1基因后对乳腺癌MDA-MB-231细胞活力的影响;应用克隆形成实验检测沉默UHRF1后对乳腺癌MDA-MB-231细胞存活的影响;吖啶橙-溴乙锭(AO/EB)检测沉默UHRF1后对乳腺癌MDA-MB-231细胞凋亡的影响;Caspase-3活性试剂盒检测沉默UHRF1后乳腺癌细胞Casapse-3活性的变化;Western blot法检测细胞中凋亡相关蛋白Bcl-2、Bax、Bad、p-Bad、XIAP、p53、p21^Cip1/Waf1和p16^INK4a的表达;应用Transwell实验研究沉默UHRF1对MDA-MB-231细胞侵袭能力的影响;Wound Healing实验研究沉默UHRF1后对其迁移能力的影响。结果沉默UHRF1使乳腺癌MDA-MB-231细胞活力降低;克隆形成实验结果显示沉默UHRF1后MDA-MB-231细胞存活能力降低,AO/EB染色显示沉默UHRF1促进MDA-MB-231细胞凋亡。同时Caspase-3活性实验结果显示沉默UHRF1后乳腺癌MDA-MB-231细胞Caspase-3的活性增加;Western blot结果显示,沉默UHRF1后,能够使凋亡蛋白Bad、XIAP和Bax的表达上调,同时抗凋亡蛋白p-Bad,Bcl-2的表达下调,也使p53,p21Cip1/Waf1,p16^INK4a蛋白表达升高;Transwell以及Wound Healing实验证明沉默UHRF1能够抑制乳腺癌MDA-MB-231细胞侵袭和迁移。结论沉默UHRF1能够抑制乳腺癌MDA-MB-231细胞活力和存活,并抑制乳腺癌MDA-MB-231的侵袭和迁移。沉默UHRF1通过调控p53,p21Cip1/Waf1,p16^INK4a信号发挥作用。

关键词: UHRF1, 乳腺癌, p53, 侵袭, 凋亡

Abstract: Objective The of this study was to investigate the effect of UHRF1 on the proliferation and metastasis of breast cancer MDA-MB-231 cells and its mechanism.Methods The effect of silencing UHRF1 gene on the viability of MDA-MB-231 cells was detected by MTT assay.Colony formation assay was performed to analyze the effect of silencing UHRF1 on cell survival of MDA-MB-231 cells.The effect of silencing UHRF1 on the apoptosis of MDA-MB-231 cells was detected by acridine orange-ethidium bromide (AO / EB).Caspase-3 activity kit was used to detect the expression of caspase-3 in MDA-MB-231 cells.The expressions of Bcl-2,Bax,Bad,p-Bad,XIAP,p53,p21^Cip1/Waf and p16^INK4a were detectedby Western blot.The abilities of invasion and migration of MDA-MB-231 cells silenced by UHRF1were examined by Transwell and Wound healing assays,respectively.Results Silencing UHRF1 significantlydecreased the viability of MDA-MB-231 cells.Silencing UHRF1 decreased colony formation in MDA-MB-231 cells.Depletion of UHRF1 resulted in apoptosis inducedin MDA-MB-231 cells,showing nuclear morphological changes by AO/EB staining and increasing caspase-3 activity.After knockdown of UHRF1,the expression of Bad,XIAP,Bax,p53,p21^Cip1/Waf1 and p16^INK4a was up-regulatedand down-regulated the expression of p-Bad and Bcl-2 in MDA-MB-231 cells.Transwell and wound healing assays demonstrated that silencing UHRF1 could decrease metastasisin MDA-MB-231 cells.Conclusion Silencing UHRF1 can inhibit the viability and survival of MDA-MB-231 cells,and inhibit the invasion and migration of MDA-MB-231 cells regulated by p53/p21^Cip1/Waf1/p16^INK4asignalings.

Key words: UHRF1, Breast cancer, p53, Metastasis, Apoptosis

中图分类号:

R737.9

陈琢, 孔艺然. 沉默UHRF1对乳腺癌细胞增殖和转移的影响[J]. 实用肿瘤学杂志, 2017, 31(6): 512-518.

CHEN Zhuo, KONG Yiran. Effect of silencing UHRF1 on proliferation and metastasis of breast cancer cells[J]. PRACTICAL ONCOLOGY JOURNAL, 2017, 31(6): 512-518.

参考文献

1 Rao N,Qiu J,Wu J,et al.Significance of tumor-infiltrating lymphocytes and the expression of topoisomerase IIalpha in the prediction of the clinical outcome of patients with triple-negative breast cancer after taxane-anthracycline-based neoadjuvant chemotherapy[J],Chemotherapy,2017,62(2):246-255.
4 Joseph FJ,van Oepen A,Friebe M,et al.Breast sentinel lymph node biopsy with imaging towards minimally invasive surgery[J],Biomed Tech(Berl),2017[Epub ahead of print].
5 Treitl D,Radkani P,Rizer M,et al.Adenoid cystic carcinoma of the breast,20 years of experience in a single center with review of literature[J],Breast Cancer,2017[Epub ahead of print].
6 Hopfner R,Mousli M,Jeltsch JM,et al.ICBP90,a novel human CCAAT binding protein,involved in the regulation of topoisomerase IIalpha expression[J],Cancer Res,2000,60(1):121-128.
7 Abu-Alainin W,Gana T,Liloglou T,et al.UHRF1 regulation of the keap1-nrf2 pathway in pancreatic cancer contributes to oncogenesis[J],J Pathol,2016,238(3):423-433.
8 Saidi S,Popov Z,Janevska V,et al.Overexpression of UHRF1 gene correlates with the major clinicopathological parameters in urinary bladder cancer[J].Int Braz J Urol,2017,43(2):224-229.
9 Li XL,Xu JH,Nie JH,et al.Exogenous expression of UHRF1 promotes proliferation and metastasis of breast cancer cells[J].Oncol Rep,2012,28(1):375-383.
10 Bonapace IM,Latella L,Papait R,et al.Np95 is regulated by E1A during mitotic reactivation of terminally differentiated cells and is essential for S phase entry[J].J Cell Biol,2002,157(6):909-914.
11 Subramani R,Gangwani L,Nandy SB,et al.Emerging roles of microRNAs in pancreatic cancer diagnosis,therapy and prognosis(Review)[J].Int J Oncol,2015,47(4):1203-1210.
12 Kang SM,Lee HJ.MicroRNAs in human lung cancer[J].Exp Biol Med(Maywood),2014,239(11):1505-1513.
13 Ahmed K,Tabuchi Y,Kondo T.Hyperthermia:an effective strategy to induce apoptosis in cancer cells[J].Apoptosis,2015,20(11):1411-1419.
14 Chen WM,Huang MD,Sun DP,et al.Long intergenic non-coding RNA 00152 promotes tumor cell cycle progression by binding to EZH2 and repressing p15 and p21 in gastric cancer[J].Oncotarget,2016,7(9):9773-9987.
15 Li J,Zhang N,Zhang R,et al.CDC5L Promotes hTERT expression and colorectal tumor growth[J].Cell PhysiolBiochem,2017,41(6):2475-2488.
16 Seo JS,Choi YH,Moon JW,et al.Hinokitiol induces DNA demethylation via DNMT1 and UHRF1 inhibition in colon cancer cells[J].BMC Cell Biol,2017,18(1):14.
17 Li J,Zhang N,Zhang R,et al.CDC5L Promotes hTERT expression and colorectal tumor growth[J].Cell Physiol Biochem,2017,41(6):2475-2488.
18 Huang L,Li A,Liao G,et al.Curcumol triggers apoptosis of p53 mutant triple-negative human breast cancer MDA-MB 231 cells via activation of p73 and PUMA[J].Oncol Lett,2017,14(1):1080-1088.
19 Cao Y,Lin M,Bu Y,et al.p53-inducible long non-coding RNA PICART1 mediates cancer cell proliferation and migration[J].Int J Oncol,2017,50(5):1671-1682.

[1]	王晓雅,肖斌,廖扬,唐荣芝,孙朝晖,李林海. CXCR家族蛋白在不同乳腺癌亚型中的表达及临床预后分析[J]. 实用肿瘤学杂志, 2019, 33(3): 206-210.
[2]	秦文,赵楠楠,韩永焕,徐亚君. ZNF436在胃癌中的表达及调控WNT/β-catenin对胃癌细胞迁移及侵袭的影响[J]. 实用肿瘤学杂志, 2019, 33(3): 222-227.
[3]	刘志强,褚艳杰,刘静,王彦博. m白术内酯Ⅰ通过TLR4/MyD88通路调控肺癌A549细胞增殖侵袭的研究[J]. 实用肿瘤学杂志, 2019, 33(3): 228-232.
[4]	李烁,张楠楠,李亮亮,龙志平,尹慧慧,赵亚双,王帆. 外周血白细胞PDE4C基因甲基化及环境因素交互作用与乳腺癌发病的关系[J]. 实用肿瘤学杂志, 2019, 33(3): 233-238.
[5]	李嘉琪,栾瑾微,张,玉,李香兰. 763例三阴性乳腺癌临床病理特征及复发、转移影响因素分析[J]. 实用肿瘤学杂志, 2019, 33(3): 244-249.
[6]	刘佳音,燕飞虎,张艳桥. 凋亡基因Caspase3和Caspase7单核苷酸多态性与胃癌遗传易感性研究[J]. 实用肿瘤学杂志, 2019, 33(3): 250-255.
[7]	陈艳波, 孔德嘉, 张金锋, 马志刚. 乳腺癌患者CYP2D6基因多态性与他莫昔芬代谢相关性研究[J]. 实用肿瘤学杂志, 2019, 33(2): 110-114.
[8]	孙江宏, 郝明珠, 姜丹, 国飞, 李晓梅. 乳腺炎、良性增生及乳腺癌FFDM鉴别诊断思路[J]. 实用肿瘤学杂志, 2019, 33(2): 139-141.
[9]	马志杰, 王帆, 赵亚双. 乳腺癌化疗患者生存质量及影响因素分析[J]. 实用肿瘤学杂志, 2019, 33(2): 149-154.
[10]	马质莹, 尚乃舰, 蔡妙田, 沈阳, 姜智允. 孕激素及雌激素受体阳性乳腺癌HER-2受体与钼靶影像学特征及临床病理学特征的相关性分析[J]. 实用肿瘤学杂志, 2019, 33(1): 47-51.
[11]	施俊, 倪小兵, 毛敏, 彭兴春. Nultin-3联合顺铂对口腔鳞状细胞癌细胞的抑制作用及机制研究[J]. 实用肿瘤学杂志, 2018, 32(6): 488-492.
[12]	卞秀森, 李光, 关欣宇, 张伊, 狄畅, 马璨. UHRF1通过调控细胞自噬抑制肺腺癌细胞增殖的分子机制研究[J]. 实用肿瘤学杂志, 2018, 32(6): 498-502.
[13]	宋玲, 付琼. 紫云英苷通过抑制HIF-1α诱导的糖酵解途径发挥抗卵巢肿细胞作用的研究[J]. 实用肿瘤学杂志, 2018, 32(6): 503-509.
[14]	李万祯, 殷俊, 李萍, 林晓辉, 张国前, 张书旭. iR-17在非小细胞肺癌放射抗拒细胞中的表达及作用研究[J]. 实用肿瘤学杂志, 2018, 32(5): 391-397.
[15]	何煦, 叶尚勉, 董丹丹, 李科. HLA-G在乳腺癌中的表达及意义[J]. 实用肿瘤学杂志, 2018, 32(5): 410-414.