泛素特异性蛋白酶39对肺癌细胞的生物学影响

卢迪; 翟今朝; 葛祥伟; 周鑫; 武鹏; 马志强; 汪进良

doi:10.3969/j.issn.2095-5227.2023.01.010

泛素特异性蛋白酶39对肺癌细胞的生物学影响

Biological effects of ubiquitin-specific protease USP39 on lung cancer cells

摘要

摘要:
背景肺癌是世界上最常见的恶性肿瘤之一，也是死亡率最高的肿瘤。与大多数肿瘤相比，肺癌的治疗现状仍不容乐观，严重威胁着人类的健康。蛋白质泛素化是翻译后修饰之一，以不同的方式调节许多的细胞过程。泛素特异性蛋白酶39(ubiquitin specific protease 39，USP39)与多种肿瘤的发生发展密切相关，但其在肺癌中的作用机制仍有待探索。
目的构建USP39稳定过表达的人肺癌A549细胞系，观察USP39过表达对肺癌A549细胞的生物学影响。
方法采用慢病毒感染法构建USP39稳定过表达的A549 USP39细胞系，并用Western blot进行验证。利用克隆形成实验检测USP39过表达对A549细胞增殖的影响；利用划痕实验检测USP39过表达对A549细胞迁移能力的影响。
结果 Western blot结果显示，A549 USP39稳定过表达的A549细胞系构建成功。与对照组相比，克隆形成实验表明USP39稳定过表达可明显促进细胞增殖，划痕实验显示USP39过表达可显著增加A549细胞的迁移能力，且差异有统计学意义(P＜0.05)。
结论本实验成功构建了USP39稳定过表达的A549细胞系，并初步验证USP39过表达与肺癌细胞的增殖和迁移相关。

Abstract:
Background Lung cancer is one of the most common malignant tumors in the world, with the highest mortality. Compared with most tumors, the treatment of lung cancer is still not optimistic, which is a serious threat to human health. Protein ubiquitination is one of the post-translational modifications that regulate many cellular processes in different ways. Ubiquitin specific protease 39 (USP39) is closely related to the occurrence and development of a variety of tumors, but the mechanism in lung cancer remains to be explored.
Objective To construct a human lung cancer A549 cell line with stable and high expression of USP39, and observe the biological effects of high expression of USP39 on lung cancer A549 cells.
Methods Using lentivirus mediated USP39 overexpression vector, stable overexpression A549 USP39 cell line infected with lentivirus was screened by puromycin, and the results were verified by Western Blot analysis. Clone formation assay was used to detect the effect of USP39 gene overexpression on A549 cell proliferation. The effect of USP39 overexpression on the migration ability of A549 cells was detected by wound healing experiment.
Results Western blot showed that A549 USP39 overexpression stably transfected cell line was successfully constructed. Compared with the control group, the clone formation experiment showed that the stable and high expression of USP39 could significantly promote cell proliferation. Wound healing experiment showed that stable and high expression of USP39 could significantly increase the migration ability of A549 cells, and the difference was statistically significant (P < 0.05).
Conclusion The stable transfected A549 cell line with flag tag overexpression of USP39 is successfully constructed, and it is preliminarily verified that the overexpression of USP39 can significantly promote the proliferation and migration of tumor cells.

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