RNF8的亚细胞定位及其在胃癌中的表达与凋亡调控

刘健; 雷箴; 袁雯; 赵枚; 黄常志

doi:10.3971/j.issn.1000-8578.2017.17.0091

RNF8的亚细胞定位及其在胃癌中的表达与凋亡调控

Subcellular Localization of RNF8 and Its Expression and Role in Apoptosis of Gastric Cancer

摘要

摘要:
目的研究环指蛋白8（ring finger protein 8, RNF8）的亚细胞定位及其在胃癌中的表达及功能。
方法构建RNF8正常及突变的真核表达载体，转染HEK293细胞，激光共聚焦显微镜下观察其亚细胞定位情况；利用Western blot及RT-PCR法分别检测RNF8在胃癌组织及细胞中的表达情况；利用流式细胞仪检测RNF8高表达对胃癌细胞凋亡的影响。
结果野生型RNF8主要定位于胞核，其指环（RING）结构域和叉头相关（forkhead-associated domain, FHA）结构域突变均对RNF8的胞核定位有一定的影响；RNF8在胃癌细胞中的表达明显下调；外源导入RNF8可以导致胃癌MGC803细胞发生凋亡。
结论 RNF8可能通过调控细胞凋亡参与肿瘤发生发展过程。

Abstract:
Objective To investigate the subcellular localization of ring finger protein 8 (RNF8) and explore its expression and function in gastric cancer cells.
Methods The fusion eukaryotic vectors expressing wide-type or mutant EGFP-RNF8 were constructed and transfected into HEK293 cells. The subcellular localization of RNF8 was observed under laser scanning confocal microscope; Western blot and RT-PCR were used to detect RNF8 expression in gastric cancer tissues and cell lines; the role of RNF8 on gastric cancer cell apoptosis was detected using flow cytometry.
Results Wild-type RNF8 was mainly located in the nucleus, while both RING and forkhead-associated(FHA) domain mutations could partly affect nuclear localization; RNF8 showed a lower expression pattern in gastric cancer cells and its forced expression in gastric cancer cell line MGC803 led to cell apoptosis.
Conclusion RNF8 may regulate the development of gastric tumor through modulating cancer cell apoptosis.

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