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

发布时间：2018-06-13 05:28

本文选题：喉癌 + 泛素样含PHD和环指域　；参考：《第三军医大学学报》2017年13期

【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-si RNA干扰UHRF1在喉癌Hep-2细胞中的表达,采用q RT-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(P0.001)。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(P0.001)。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(P0.001)。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(P0.001)。干扰组细胞Bax蛋白表达上调(P0.001),Bcl-2蛋白表达下调(P0.001),MMP-2蛋白、MMP-9蛋白表达下调(P0.001)。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 investigate the effects of down-regulation of UHRF1 gene expression on proliferation, apoptosis, invasion and migration of Hep-2 cells by down-regulating the expression of ubiquiin-like PhD and ring finger domain 1(ubiquitin-like protein containing PhD and ring finger domains 1 (UHRF1) gene in Hep-2 cells. Methods the experiment was divided into blank group, negative control group and interference group. UHRF1-si RNA was used to interfere the expression of UHRF1 in Hep-2 cells, and the expression of UHRF1 in Hep-2 cells was detected by Q RT-PCRG Western blot. The changes of proliferation, apoptosis, invasion and migration ability of Hep-2 cells after down-regulation of UHRF1 expression were detected by flow cytometry and flow cytometry. Western blot was used to detect the changes of Bax-Bcl-2MMP-2MMP-9 protein expression in Hep-2 cells after down-regulation of UHRF1 expression. Results compared with blank group and negative control group, down-regulation of UHRF1 expression in interference group could significantly inhibit the proliferation ability of Hep-2 cells. The apoptotic rate of blank group, negative control group and interference group were 5.47 卤2.77 and 3.95 卤0.66 respectively. The apoptosis rate of interference group was significantly higher than that of control group (18.95 卤1.10). In blank group, negative control group, the number of invasive cells per visual field in the interference group was 213.7 卤13.1% and 221.3 卤10.3% respectively. The number of invasive cells in the interference group was significantly lower than that in the control group (P 0.001). In blank group, negative control group, the number of migration cells per visual field in interference group was 230.7 卤5.5 and 222.7 卤11.2 卤111.7 卤7.6, respectively. The number of migration cells in interference group was significantly lower than that in control group (P 0.001). The expression of Bax protein upregulated and down-regulated the expression of P0.001MMP 2 protein and MMP-9 protein in interference group. Conclusion down-regulation of UHRF1 expression in Hep-2 cells can significantly inhibit cell proliferation, promote cell apoptosis and inhibit cell invasion and migration.
【作者单位】：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;
【基金】：国家自然科学基金面上项目(81272980) 国家临床重点专科建设项目([2012]649)~~
【分类号】：R739.65

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