[1]文韬宇,祝琳,刘亚男,等.下调泛素样含PHD和环指域1基因的表达对喉癌Hep-2细胞增殖侵袭能力的影响[J].第三军医大学学报,2017,39(13):1339-1344.
 WEN Taoyu,ZHU Lin,LIU Yanan,et al.Effect of down-regulation of UHRF1 on proliferation and invasive capacity in human laryngeal carcinoma Hep-2 cells[J].J Third Mil Med Univ,2017,39(13):1339-1344.
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下调泛素样含PHD和环指域1基因的表达对喉癌Hep-2细胞增殖侵袭能力的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
39卷
期数:
2017年第13期
页码:
1339-1344
栏目:
基础医学
出版日期:
2017-07-15

文章信息/Info

Title:
Effect of down-regulation of UHRF1 on proliferation and invasive capacity in human laryngeal carcinoma Hep-2 cells
作者:
文韬宇祝琳刘亚男万婕雷越陈鸿雁
重庆医科大学附属第一医院耳鼻喉科;西南医科大学附属医院耳鼻咽喉头颈外科
Author(s):
WEN Taoyu ZHU Lin LIU Yanan WAN Jie LEI Yue CHEN Hongyan

Department of Otolaryngology, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016; Department of Otolaryngology-Head and Neck Surgery, Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan Province, 646000, China

关键词:
喉癌泛素样含PHD和环指域1增殖凋亡侵袭迁移
Keywords:
laryngeal carcinoma ubiquitin-like protein containing PHD and ring finger domains 1 proliferation apoptosis invasion: migration
分类号:
R73-37; R730.23; R739.65
文献标志码:
A
摘要:

目的         探讨下调泛素样含PHD和环指域1(ubiquitinlike protein containing PHD and RING finger domains 1,UHRF1)基因在喉癌Hep-2细胞中的表达后对其增殖,凋亡,侵袭迁移能力的影响。方法        实验分为空白组,阴性对照组和干扰组。 利用UHRF1-siRNA干扰UHRF1在喉癌Hep-2细胞中的表达,采用qRT-PCR、Western blot检测UHRF1的表达变化。采用MTT法、流式细胞术、Transwell实验检测下调UHRF1表达后Hep-2细胞的增殖、凋亡、侵袭、迁移能力的变化。Western blot检测下调UHRF1表达后Hep-2细胞内Bax、Bcl-2、MMP-2、MMP-9蛋白表达的变化情况。结果         与空白组和阴性对照组相比干扰组下调UHRF1的表达后可明显抑制喉癌Hep2细胞的增殖能力(P<0.001)。空白组,阴性对照组,干扰组凋亡率分别为(5.47±2.77)%,(3.95±0.66)%,(18.95±1.10)%,干扰组凋亡率明显提高(P<0.001)。空白组,阴性对照,干扰组每视野侵袭细胞数分别为(213.7±13.1),(221.3±10.3),(97.7±7.5),干扰组侵袭细胞数明显降低(P<0.001)。空白组,阴性对照,干扰组每视野迁移细胞数分别为(230.7±5.5),(222.7±11.2),(111.7±7.6),干扰组迁移细胞数明显降低(P<0.001)。干扰组细胞Bax蛋白表达上调(P<0.001),Bcl-2蛋白表达下调(P<0.001),MMP-2蛋白、MMP-9蛋白表达下调(P<0.001)。结论          在喉癌Hep-2细胞中下调UHRF1的表达后可明显抑制细胞的增殖,促进细胞凋亡,抑制细胞的侵袭迁移能力。

Abstract:

Objective         To determine the effect of down-regulation of ubiquitin-like protein containing PHD and ring finger domains 1 (UHRF1) on the proliferation, apoptosis, invasion and migration in human laryngeal carcinoma Hep-2 cell line. Methods        The Hep-2 cells were randomly divided into 3 groups: blank control group, negative control group and tranfected group. UHRF1-siRNA was used to down-regulate the expression of UHRF1 in the Hep-2 cells, and quantitative real-time PCR (qRT-PCR) and Western blotting were used to detect the mRNA and protein expression of UHRF1 after down-regulation. The proliferative, invasive and migratory capabilities of Hep-2 cells transfected with UHRF1-siRNA were detected by MTT assay, flow cytometry and Transwell chamber assay. Western blotting was used to measure the protein levels of Bcl-2, Bax, MMP-2 and MMP-9. Results        Compared with the blank control group and negative control group, down-regulation of UHRF1 significantly inhibited the proliferative capabilities of Hep-2 cells (P<0.001). The apoptotic rate was (5.47±2.77)%, (3.95±0.66)% and (18.95±1.10)%, respectively in the blank control group, negative control group and transfected group, and the rate was the highest in the transfected group. The number of invasive cells was (213.7±13.1), (221.3±10.3) and (97.7±7.5) respectively, and that of migratory cells was (230.7±5.5), (222.7±11.2) and (111.7±7.6) respectively in the above 3 groups. The numbers of invasive and migratory cells were both obviously lower in the transfected group (P<0.001). Western blotting results showed that Bax was up-regulated (P<0.001), and Bcl-2, MMP-2 and MMP-9 were down-regulated (P<0.001). Conclusion        Down-regulation of UHRF1 inhibits the proliferation, induces the apoptosis, and reduces the invasive and migratory capability in Hep-2 cells.

参考文献/References:

[1]孔维佳,周梁,许庚, 等. 耳鼻咽喉头颈外科学[M].北京:人民卫生出版社,2010:460-464.
KONG W J, ZHOU L, XU G, et al.Otorhinolaryngology-Head and Neck Surgery [M]. Beijing:People’s Medical Publishing House, 2010:460-464.
[2]BRONNER C, KRIFA M, MOUSLI M. Increasing role of UHRF1 in the reading and inheritance of the epigenetic code as well as in tumorogenesis[J]. Biochem Pharmacol, 2013, 86(12): 1643-1649. DOI:10.1016/j.bcp.2013.10.002.
[3]PI J T, LIN Y, QUAN Q, et al. Overexpression of UHRF1 is significantly associated with poor prognosis in laryngeal squamous cell carcinoma[J]. Med Oncol, 2013, 30(4): 613. DOI:10.1007/s12032-013-0613-9.
[4]JENKINS Y, MARKOVTSOV V, LANG W, et al. Critical role of the ubiquitin ligase activity of UHRF1, a nuclear RING finger protein, in tumor cell growth[J]. Mol Biol Cell, 2005, 16(12): 5621-5629. DOI:10.1091/mbc.E05-03-0194.
[5]MOUSLI M, HOPFNER R, ABBADY A Q, et al. ICBP90 belongs to a new family of proteins with an expression that is deregulated in cancer cells[J]. Br J Cancer, 2003, 89(1): 120-127. DOI:10.1038/sj.bjc.6601068.
[6]JIN W, CHEN L, CHEN Y, et al. UHRF1 is associated with epigenetic silencing of BRCA1 in sporadic breast cancer[J]. Breast Cancer Res Treat, 2010, 123(2): 359-373. DOI:10. 1007/s10549-009-0652-2.
[7]UNOKI M, DAIGO Y, KOINUMA J, et al. UHRF1 is a novel diagnostic marker of lung cancer[J]. Br J Cancer, 2010, 103(2): 217-222. DOI:10.1038/sj.bjc.6605717.
[8]UNOKI M, KELLY J D, NEAL D E, et al. UHRF1 is a novel molecular marker for diagnosis and the prognosis of bladder cancer[J]. Br J Cancer, 2009, 101(1): 98-105. DOI:10.1038/ sj.bjc.6605123.
[9]CUI L, CHEN J, ZHANG Q, et al. Up-regulation of UHRF1 by oncogenic Ras promoted the growth, migration, and metastasis of pancreatic cancer cells[J]. Mol Cell Biochem, 2015, 400(1-2): 223-232. DOI:10.1007/s11010-014-2279-9.
[10]SABATINO L, FUCCI A, PANCIONE M, et al. UHRF1 coordinates peroxisome proliferator activated receptor gamma (PPARG) epigenetic silencing and mediates colorectal cancer progression[J]. Oncogene, 2012, 31(49): 5061-5072. DOI:10. 1038/onc.2012.3.
[11]SABATINO L, FUCCI A, PANCIONE M, et al. Cell cycle and/or proliferation markers: what is the best method to discriminate cervical high-grade lesions[J]. Hum Pathol, 2005, 36(10): 1101-1107. DOI:10.1016/j.humpath.2005.07.016.
[12]HOPFNER R, MOUSLI M, JELTSCH J M, 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.
[13]MULDER K W, WANG X, ESCRIU C, et al. Diverse epigenetic strategies interact to control epidermal differentiation[J]. Nat Cell Biol, 2012, 14(7): 753-763. DOI:10.1038/ncb2520.
[14]WANG F, YANG Y Z, SHI C Z, et al. UHRF1 promotes cell growth and metastasis through repression of p16(inka) in colorectal cancer[J]. Ann Surg Oncol, 2012, 19(8): 2753-2762. DOI:10.1245/s10434-011-2194-1.
[15]QIN Y, WANG J, GONG W, et al. UHRF1 depletion suppresses growth of gallbladder cancer cells through induction of apoptosis and cell cycle arrest[J]. Oncol Rep, 2014, 31(6): 2635-2643. DOI:10.3892/or.2014.3145.

更新日期/Last Update: 2017-07-12