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1.400016 重庆,重庆医科大学附属第一医院耳鼻喉科
2.646000 四川 泸州,西南医科大学附属医院耳鼻咽喉头颈外科
[ 陈鸿雁,E-mail: chenhongyan1963@hotmail.com CHEN Hongyan,E-mail:chenhongyan1963@hotmail.com
收稿:2016-12-18,
修回:2017-02-12,
纸质出版:2017-07-15
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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.
Taoyu WEN, Lin ZHU, Yanan LIU, et al. Effect of down-regulation of UHRF1 on proliferation and invasive capacity in human laryngeal carcinoma Hep-2 cells[J]. Journal of Third Military Medical University, 2017, 39(13): 1339-1344.
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. DOI: 10.16016/j.1000-5404.201612122.
Taoyu WEN, Lin ZHU, Yanan LIU, et al. Effect of down-regulation of UHRF1 on proliferation and invasive capacity in human laryngeal carcinoma Hep-2 cells[J]. Journal of Third Military Medical University, 2017, 39(13): 1339-1344. DOI: 10.16016/j.1000-5404.201612122.
目的
2
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
2
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
2
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
2
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。
Objective
2
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
2
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
2
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 w
as 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
2
Down-regulation of UHRF1 inhibits the proliferation
induces the apoptosis
and reduces the invasive and migratory capability in Hep-2 cells.
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