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

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UHRFl-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.

Effect of down-regulation of UHRF1 on proliferation and invasive capacity in human laryngeal carcinoma Hep-2 cells

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.