RNA干扰UHRF1基因表达对食管癌细胞生物学行为的影响

Effects of UHRF1 gene silencing by RNA interference on biological behaviors of esophageal carcinoma cells

目的:通过短发夹RNA(short hairpin RNA,shRNA)干扰UHRF1(ubiquitin-like with PHD and ring finger domains 1)基因表达,探讨沉默UHRF1基因对食管癌TE-1细胞生物学行为的影响。方法:采用免疫组织化学法检测食管癌组织及正常食管组织中UHRF1蛋白的表达水平,以证实UHRF1在食管癌组织中过表达;以慢病毒感染的方法将UHRF1基因的shRNA转入食管癌TE-1细胞株,采用RT-PCR和蛋白质印迹法检测转染UHRF1-shRNA重组表达载体后TE-1细胞中UHRF1mRNA和蛋白的表达水平;并分别以MTT法、FCM法和体外侵袭实验检测UHRF1基因沉默对TE-1细胞增殖、周期、凋亡和侵袭能力的影响。结果:UHRF1在正常食管组织中不表达或低表达,在食管癌组织中过表达(P<0.01);UHRF1-shRNA转染后,TE-1细胞中UHRF1mRNA和蛋白表达水平显著降低;UHRF1基因沉默可使TE-1细胞增殖及体外侵袭能力明显下降,G0/G1期细胞增多,凋亡增加(P<0.05)。结论:沉默UHRF1基因可降低食管癌细胞的增殖和侵袭能力,促进细胞凋亡,提示UHRF1在食管癌发生和发展过程中发挥重要作用。

Objective： TO investigate the effects of UHRF1 （ubiquitin-like with PHD and ring finger domains 1） gene silencing by shRNA （short hairpin RNA） interference on the biological behaviors of esophageal carcinoma TE-1 cells. Methods： The protein expression levels of UHRF1 in esophageal carcinoma tissues and normal esophageal tissues were detected by immunohistochemical staining assay to confirm overexpression of UHRF1 in esophageal carcinoma tissues. After UHRFI-shRNA was transfected into TE-1 cells by method of lentiviral infection, the mRNA and protein levels of UHRF1 in TE-1 cells were detected by RT-PCR （reverse transcription-PCR） and Western blotting, respectively. The proliferative and invasive capabilities of TE-1 cells transfected with UHRFI-shRNA were detected by the MTT assay and Transwell invasion assay, respectively. The cell cycle distribution and apoptosis were determined by FCM （flow cytometry）. Results： UHRF1 was overexpressed in esophageal carcinoma tissues, as compared with the normal esophageal tissues （low or absent expression, P 〈 0.01）. After transfection with UHRFI-shRNA, the mRNA and protein levels of UHRF1 in TE-1 cells were significantly reduced, the proliferative and invasive capabilities of TE-1 cells were inhibited, and the cell cycle was arrested in G0/G1 phase （P 〈 0.05） with an increase in apoptosis （P 〈 0.05）. Conclusion： Small RNA interference targeting UHRF1 gene can inhibit the proliferation, reduce the invasive capability, and induce the apoptosis of TE-1 cells, indicating that UHRF1 plays an important role in oncogenesis and progression of human esophageal carcinoma.