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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(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。
Objective To investigate the effects of down-regulation of ubiquitin-like protein containing PHD and RING finger domains 1 (UHRF1) gene on proliferation, apoptosis, invasion and migration of laryngeal carcinoma Hep-2 cells Impact. 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 laryngeal carcinoma Hep-2 cells. The expression of UHRF1 was detected by q RT-PCR and Western blot. MTT assay, flow cytometry and Transwell assay were used to detect the changes of proliferation, apoptosis, invasion and migration of Hep-2 cells after UHRF1 expression was down-regulated. Western blot was used to detect the expression of Bax, Bcl-2, MMP-2 and MMP-9 in Hep-2 cells after UHRF1 down-regulation. Results Compared with blank group and negative control group, the downregulation of UHRF1 expression in Hep-2 cells inhibited the proliferation of Hep-2 cells (P <0.001). The apoptotic rate in the blank group, negative control group and interference group were (5.47 ± 2.77)%, (3.95 ± 0.66)% and (18.95 ± 1.10)%, respectively. The apoptosis rate in the interference group was significantly increased (P <0.001). The number of invasive cells in each group was (213.7 ± 13.1), (221.3 ± 10.3) and (97.7 ± 7.5) in the blank group and negative control group, respectively. The number of invasive cells in the interference group was significantly decreased (P <0.001). The number of migrating cells per field of view in the blank group and negative control group was (230.7 ± 5.5), (222.7 ± 11.2) and (111.7 ± 7.6), respectively. The number of migrating cells in the interference group was significantly decreased (P <0.001). Bcl-2 protein expression was down-regulated (P <0.001) and MMP-2 protein and MMP-9 protein expression were down-regulated in the intervention group (P <0.001). Conclusion Down-regulation of UHRF1 expression in laryngeal carcinoma Hep-2 cells can significantly inhibit cell proliferation, promote apoptosis and inhibit the invasion and migration of cells.