| 引用本文： |
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常换换,李 姣,贾 松,等.TMEFF2基因对人胚肾细胞生物学行为的影响[J].同济大学学报（医学版）,2019,40(6):801-808. [点击复制]
- CHANG Huan-huan,LI Jiao,,JIA Song,et al.Effect of TMEFF2 gene on biological behavior of human embryonic kidney cells[J].同济大学学报（医学版）,2019,40(6):801-808. [点击复制]
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| 摘要: |
| 目的 探究TMEFF2基因对人胚肾细胞生物学行为的影响及可能的机制。方法 构建基因低表达载体shRNA-TMEFF2,将实验组shRNA-TMEFF2、对照组NC-TMEFF2的DNA分别转染进入HEK293细胞；采用qPCR和Western印迹法检测TMEFF2的表达；用CCK-8检测细胞增殖能力；流式细胞术检测细胞周期和凋亡率；用RNA-seq分析TMEFF2基因低表达对HEK293细胞基因表达谱的影响；划痕法和Transwell法检测细胞迁移、侵袭能力的变化。结果 实验组中mRNA及蛋白水平明显降低；细胞增殖能力显著降低；细胞周期阻滞在G1期且凋亡率增高；细胞迁移、侵袭的能力均受到抑制，差异均具有统计学意义(P均<0.05)；RNA-seq显示实验组共有4769个基因表达差异有统计学意义，其中2355个基因上调，占49.4%，2414个基因下调，占50.6%。结论 TMEFF2基因低表达能抑制HEK293细胞的增殖，抑制HEK293细胞迁移、侵袭的能力，可能促进细胞的凋亡，其作用机制可能与RGCC及ADGRL1-FLRT信号通路有关。 |
| 关键词: TMEFF2基因 HEK293 增殖 细胞周期 凋亡 迁移 |
| DOI：10.16118/j.1008-0392.2019.06.007 |
| 投稿时间：2019-02-22 |
| 基金项目:国家重点基础研究发展计划（973计划）（2012CBA01308） |
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| Effect of TMEFF2 gene on biological behavior of human embryonic kidney cells |
| CHANG Huan-huan,LI Jiao,,JIA Song,XU Jie,,Lü Li-Xia,XU Lei |
| (Dept. of Biochemistry and Molecular Biology Research, Regenerative Medicine, Tongji University School of Medicine, Shanghai 200092, China) |
| Abstract: |
| Objective To investigate the effect of TMEFF2 gene on the biological behavior of human embryonic kidney cells and its possible mechanism. Methods Human embryonic kidney HEK293 cells were transfected with shRNA-TMEFF2（experimental group) and NC-TMEFF2（control group) vectors, respectively. The expression of TMEFF2 mRNA and protein was detected by qPCR and Western blotting, respectively; the proliferation ability was detected by CCK-8; the cell cycle and apoptosis rate were detected by flow cytometry; the cell migration and invasion ability were detected by scratch test and Transwell assay. The effect of low expressed TMEFF2 gene on expression profile of HEK293 cells was analyzed by RNA-seq. Results In the experimental group, the TMEFF2 mRNA and protein expression levels were significantly decreased; the cell proliferation ability was significantly reduced; the cell cycle was blocked in the G1 phase; the apoptotic rate was higher; the cell migration and invasion ability were inhibited, compared with the control group（P<0.05). RNA-seq showed that there were 4769 genes differentially expressed in the experimental group, of which 2355 genes were up-regulated（49.4%) and 2414 genes were down-regulated（50.6%). Conclusion Low expression of TMEFF2 gene can inhibit the proliferation, migration and invasion of HEK293 cells, and promote cell apoptosis. The mechanism may be related to RGCC and ADGRL1-FLRT signaling pathway. |
| Key words: TMEFF2 gene HEK293 proliferation cell cycle apoptosis migration |
