Effects of silencing the expression of UBE2T gene on proliferation, apoptosis, migration and invasion abilities of lung cancer A549 cells_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

 CHEN Hongyun , LI Xiaoming , HUANG Guosheng , YANG Jinhua , QIAO Fei

Second ward of general thoracic surgery, The First Affiliated Hospital of Nanyang Medical College, Nanyang, Henan 473000, P. R. China;

Corresponding author：

CHEN Hongyun, Email: chenhongyun_88@163.com

Keywords：

Non-small cell lung cancer; Ubiquitin-conjugating enzyme E2T; Proliferation; Invasion; Migration

DOI：

10.7507/1671-6205.202011058

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To explore the effects of silencing the expression of ubiquitin-conjugating enzyme E2T (UBE2T) gene on proliferation, apoptosis, migration and invasion abilities of A549 cells.Methods A549 cells were cultured in vitro. Three sets of shRNA-UBE2T plasmid vectors (UBE2T-shRNA1 group, UBE2T-shRNA2 group, UBE2T-shRNA3 group) and shRNA-NC (shRNA-NC group) were constructed, respectively. A549 cells were transfected with lipofection transfection. The cells transfected with empty vector were enrolled as control (control group). The transfection efficiency was detected by RT-PCR. The effects of silencing the expression of UBE2T gene on biological behaviors (proliferation, apoptosis, migration, and invasion) of lung cancer A549 cells were detected by clone formation assay, flow cytometry, Transwell assay and scratch test. The expression of proliferation and apoptosis related proteins, and expression of PI3K/AKT signaling pathway proteins were detected by Western blot. Results After transfection, expression level of UBE2T mRNA in UBE2T-shRNA1 group, UBE2T-shRNA2 group and UBE2T-shRNA3 group was significantly down-regulated (all P<0.05), whose down-regulation was the most significant in UBE2T-shRNA3 group (P<0.05). Compared with control group and shRNA-NC group, clone formation rate, number of invasion A549 cells, scratch healing rate, Ki67 expression, PCNA expression, p-PI3K/PI3K ratio and p-AKT/AKT ratio were significantly decreased in UBE2T-shRNA3 group (P<0.05), while A549 apoptosis rate, Bax/Bcl-2 ratio and cleaved caspase-3/caspase-3 ratio were significantly increased (P<0.05). There were no significant differences in the above indexes between control group and shRNA-NC group (P>0.05). Conclusions The shRNA interfering with UBE2T is reliable to construct the model of A549 cells with stable low-expression UBE2T. Down-regulation of UBE2T expression can promote apoptosis of A549 cells, inhibit their proliferation, invasion and migration abilities. The mechanism may be related to inhibiting the activation of PI3K/AKT signaling pathway.

Citation： CHEN Hongyun, LI Xiaoming, HUANG Guosheng, YANG Jinhua, QIAO Fei. Effects of silencing the expression of UBE2T gene on proliferation, apoptosis, migration and invasion abilities of lung cancer A549 cells. Chinese Journal of Respiratory and Critical Care Medicine, 2021, 20(8): 577-583. doi: 10.7507/1671-6205.202011058 Copy

1.	Al MB, Brennan PC, Mellothoms C. A review of lung cancer screening and the role of computer-aided detection. Clin Radiol, 2017, 72(6): 433-442.
2.	薛英杰, 贾靖, 吴杨, 等. 吉西他滨与培美曲塞分别联合顺铂治疗非小细胞肺癌的临床效果. 解放军医药杂志, 2018, 30(12): 11-14.
3.	Morreale FE, Bortoluzzi A, Chaugule VK, et al. Allosteric targeting of the Fanconi anemia ubiquitin-conjugating enzyme Ube2T by fragment screening. J Med Chem, 2017, 60(9): 4093-4098.
4.	刘瑞廷, 侯亚莉, 白继荣, 等. UBE2T 基因通过调控 ROS 抑制结直肠癌细胞凋亡. 现代肿瘤医学, 2019, 27(12): 2075-2078.
5.	李伟, 郭梓鑫, 李永红, 等. 利用 GEO 数据集分析 UBE2T 在肺癌中的表达及临床意义. 肿瘤代谢与营养电子杂志, 2019, 6(2): 242-245.
6.	Yu H, Xiang P, Pan Q, et al. Ubiquitin-conjugating enzyme E2T is an independent prognostic factor and promotes gastric cancer progression. Tumor Biol, 2016, 37(9): 11723-11732.
7.	Yu H, Kim YM, Cho M. Cytoplasm-localized sirt1 downregulation attenuates apoptosis and cell cycle arrest in cisplatin-resistant lung cancer A549 cells. J Cancer, 2020, 11(15): 4495-4509.
8.	Rickman K, Lach F, Abhyankar A, et al. Deficiency of UBE2T, the E2 ubiquitin ligase necessary for FANCD2 and FANCI ubiquitination, causes FA-T subtype of Fanconi anemia. Cell Rep, 2015, 12(1): 35-41.
9.	Ueki T, Park JH, Nishidate T, et al. Ubiquitination and downregulation of BRCA1 by ubiquitin-conjugating enzyme E2T overexpression in human breast cancer cells. Cancer Res, 2009, 69(22): 8752-8760.
10.	Puisieux A, Pommier RM, Morel AP, et al. Cellular pliancy and the multistep process of tumorigenesis. Cancer Cell, 2018, 33(2): 164-172.
11.	Hao J, Xu A, Xie X, et al. Elevated expression of UBE2T in lung cancer tumors and cell lines. Tumour Biol, 2008, 29(3): 195-203.
12.	Fritz M, Lima EABF, Nikolić V, et al. Local and nonlocal phase-field models of tumor growth and invasion due to ECM degradation. Mathemat Models Methods Applied Sci, 2019, 29(13): 2433-2468.
13.	Nargis NN, Aldredge RC, Guy RD. The influence of soluble fragments of extracellular matrix (ECM) on tumor growth and morphology. Mathemat Biosci, 2018, 296: 1-16.
14.	Wen MX, Kwon YK, Wang YX, et al. Elevated expression of UBE2T exhibits oncogenic properties in human prostate cancer. Oncotarget, 2015, 6(28): 25226-25239.
15.	赵勇, 朱建国, 单显民, 等. 土木香内酯增强胶质瘤干细胞对化疗敏感性的分子机制研究. 解放军医药杂志, 2019, 31(11): 15-19.
16.	Wang H, Yu Z, Huo S, et al. Overexpression of ELF3 facilitates cell growth and metastasis through PI3K/Akt and ERK signaling pathways in non-small cell lung cancer. Int J Biochem Cell Biol, 2018, 94: 98-106.
17.	Pal A, Young MA, Donato NJ. Emerging potential of therapeutic targeting of ubiquitin-specific proteases in the treatment of cancer. Cancer Res, 2014, 74(18): 4955-4966.

1. Al MB, Brennan PC, Mellothoms C. A review of lung cancer screening and the role of computer-aided detection. Clin Radiol, 2017, 72(6): 433-442.
2. 薛英杰, 贾靖, 吴杨, 等. 吉西他滨与培美曲塞分别联合顺铂治疗非小细胞肺癌的临床效果. 解放军医药杂志, 2018, 30(12): 11-14.
3. Morreale FE, Bortoluzzi A, Chaugule VK, et al. Allosteric targeting of the Fanconi anemia ubiquitin-conjugating enzyme Ube2T by fragment screening. J Med Chem, 2017, 60(9): 4093-4098.
4. 刘瑞廷, 侯亚莉, 白继荣, 等. UBE2T 基因通过调控 ROS 抑制结直肠癌细胞凋亡. 现代肿瘤医学, 2019, 27(12): 2075-2078.
5. 李伟, 郭梓鑫, 李永红, 等. 利用 GEO 数据集分析 UBE2T 在肺癌中的表达及临床意义. 肿瘤代谢与营养电子杂志, 2019, 6(2): 242-245.
6. Yu H, Xiang P, Pan Q, et al. Ubiquitin-conjugating enzyme E2T is an independent prognostic factor and promotes gastric cancer progression. Tumor Biol, 2016, 37(9): 11723-11732.
7. Yu H, Kim YM, Cho M. Cytoplasm-localized sirt1 downregulation attenuates apoptosis and cell cycle arrest in cisplatin-resistant lung cancer A549 cells. J Cancer, 2020, 11(15): 4495-4509.
8. Rickman K, Lach F, Abhyankar A, et al. Deficiency of UBE2T, the E2 ubiquitin ligase necessary for FANCD2 and FANCI ubiquitination, causes FA-T subtype of Fanconi anemia. Cell Rep, 2015, 12(1): 35-41.
9. Ueki T, Park JH, Nishidate T, et al. Ubiquitination and downregulation of BRCA1 by ubiquitin-conjugating enzyme E2T overexpression in human breast cancer cells. Cancer Res, 2009, 69(22): 8752-8760.
10. Puisieux A, Pommier RM, Morel AP, et al. Cellular pliancy and the multistep process of tumorigenesis. Cancer Cell, 2018, 33(2): 164-172.
11. Hao J, Xu A, Xie X, et al. Elevated expression of UBE2T in lung cancer tumors and cell lines. Tumour Biol, 2008, 29(3): 195-203.
12. Fritz M, Lima EABF, Nikolić V, et al. Local and nonlocal phase-field models of tumor growth and invasion due to ECM degradation. Mathemat Models Methods Applied Sci, 2019, 29(13): 2433-2468.
13. Nargis NN, Aldredge RC, Guy RD. The influence of soluble fragments of extracellular matrix (ECM) on tumor growth and morphology. Mathemat Biosci, 2018, 296: 1-16.
14. Wen MX, Kwon YK, Wang YX, et al. Elevated expression of UBE2T exhibits oncogenic properties in human prostate cancer. Oncotarget, 2015, 6(28): 25226-25239.
15. 赵勇, 朱建国, 单显民, 等. 土木香内酯增强胶质瘤干细胞对化疗敏感性的分子机制研究. 解放军医药杂志, 2019, 31(11): 15-19.
16. Wang H, Yu Z, Huo S, et al. Overexpression of ELF3 facilitates cell growth and metastasis through PI3K/Akt and ERK signaling pathways in non-small cell lung cancer. Int J Biochem Cell Biol, 2018, 94: 98-106.
17. Pal A, Young MA, Donato NJ. Emerging potential of therapeutic targeting of ubiquitin-specific proteases in the treatment of cancer. Cancer Res, 2014, 74(18): 4955-4966.

Previous Article
Protection of resveratrol on seawater-drowning-induced lung injury in rats
Next Article
尼达尼布治疗放射性肺损伤一例并文献复习

Chinese Journal of Respiratory and Critical Care Medicine

Effects of silencing the expression of UBE2T gene on proliferation, apoptosis, migration and invasion abilities of lung cancer A549 cells

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content