南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (9): 1329-1333.doi: 10.12122/j.issn.1673-4254.2021.09.06

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ELF4通过激活Akt通路促进胰岛素瘤细胞增殖并抑制细胞凋亡

卫国红，王 利，万学思，谭泳谣   

  1. 中山大学附属第一医院内分泌科，保健科，广东 广州 510080；中山大学中山医学院，广东 广州 510080
  • 出版日期:2021-09-20 发布日期:2021-09-30

ELF4 promotes proliferation and inhibits apoptosis of human insulinoma cells by activating Akt signaling

WEI Guohong, WANG Li, WAN Xuesi, TAN Yongyao   

  1. Department of Endocrinology, Department of Healthcare, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China; Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China
  • Online:2021-09-20 Published:2021-09-30

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摘要： 目的 探索ELF4促进胰岛素瘤细胞增殖与抗凋亡的作用及其机理。方法 利用逆转录病毒载体系统，构建稳定高表达ELF4的胰岛素瘤细胞BON-ELF4，以及载体对照组细胞BON-Vector。采用MTT法检测胰岛素瘤细胞BON-ELF4和BON-Vector各组细胞的生长情况。采用0.1 μmol/L化疗药物表阿霉素处理BON-ELF4和BON-Vector细胞24 h后，检测细胞凋亡相关指标，采用免疫印迹法检测各组肿瘤细胞中凋亡通路关键因子caspase-9，caspase-8和PARP切割带的表达水平；采用Annexin V-FITC/PI双染色结合流式细胞术分析各组细胞凋亡情况。采用免疫印迹法检测胰岛素瘤细胞BON-ELF4和BON-Vector细胞中Akt磷酸化水平。结果 免疫印迹法检测结果显示，BON-ELF4细胞中ELF4蛋白表达量高于对照组细胞BON-Vector（P=0.013），稳定高表达外源性ELF4蛋白的细胞系BON-ELF4构建成功。稳定高表达ELF4的细胞系BON-ELF4生长明显较快（P<0.05）。相比较于载体对照组 BON-Vector，BON-ELF4 细胞在 0.1 μmol/L 表阿霉素处理 24 h 后 caspase-8，caspase-9的蛋白前体降低，而caspase-8，caspase-9切割成熟体升高（P<0.05）；同时，相应的PARP切割带随着ELF4的高表达而降低（P<0.05）；流式细胞术检测结果显示，22.90%的载体对照组BON-Vector细胞呈现出Annexin V阳性、PI阴性的早期凋亡结果，6.03%的BON-ELF4细胞组呈现出Annexin V阳性、PI阴性的早期凋亡结果。外源高表达ELF4的肿瘤细胞中Akt的磷酸化显著升高（P<0.05），而Akt总蛋白的水平基本未受影响（P>0.05）。结论 ELF4能通过激活Akt通路而促进胰岛素瘤胞增殖和抗凋亡。

关键词: ELF4；胰岛素瘤细胞；Akt信号通路

Abstract: Objective To investigate the effect of overexpression of the oncogenic transcription factor ELF4 on proliferation and apoptosis in human insulinoma cells and explore the underlying mechanism. Methods A human insulinoma BON cell line with stable overexpression of ELF4 (BON-ELF4 cells) was constructed using a recombinant retrovirus vector and the expression of ELF4 protein was verified using Western blotting. MTT assay was used to assess the proliferation of BON-ELF4 cells and BON-Vector cells, and the cell apoptosis induced by treatment with epirubicin (0.1 μmol/L for 24 h) was analyzed by detecting the expressions of cleaved caspase-8, caspase-9, and PARP using Western blotting. Flow cytometry with Annexin V-FITC/PI staining was performed to analyze the numbers of apoptotic BON-Vector or BON-ELF4 cells. The expressions of phosphorylated Akt and total Akt in the cells were detected using Western blotting. Results BON-ELF4 cell line with stable overexpression of ELF4 was successfully established. ELF4 overexpression significantly promoted the proliferation (P<0.05) and obviously suppressed epirubicin-induced apoptosis in BON cells, resulting also in significantly reduced expressions of cleaved caspase-8, caspase-9 and PARP (P<0.05). The results of flow cytometry showed a significantly lower apoptotic rate in BON-ELF4 cells than in BON-Vector cells following epirubicin treatment (6.03% vs 22.90%). The phosphorylation levels of Akt (Thr308 and Ser473) were significantly increased (P<0.05) while the level of total Akt remained unchanged (P>0.05) in ELF4-overexpressing cells. Conclusion ELF4 overexpression enhances the proliferation and suppresses apoptosis of insulinomas cells by activating Akt signaling.

Key words: ELF4; insulinoma cells; Akt signaling pathway