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[蛋白精氨酸甲基转移酶1通过促进核糖核苷酸还原酶M2亚基表达抑制鼻咽癌细胞凋亡]

[PRMT1 inhibits apoptosis of nasopharyngeal carcinoma cells by promoting RRM2 expression].

作者信息

Zhou L, Wu J, Zhang M, Zhao B, Ma S

机构信息

Department of Otorhinolaryngology-Head and Neck Surgery, First Affiliated Hospital of Bengbu Medical College, Bengbu 233000, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2022 Dec 20;42(12):1783-1790. doi: 10.12122/j.issn.1673-4254.2022.12.05.

Abstract

OBJECTIVE

To verify whether PRMT1 inhibits apoptosis of nasopharyngeal carcinoma (NPC) cells by promoting RRM2 expression.

METHODS

Immunohistochemistry and Western blotting were performed to detect the relative expression of PRMT1 and RRM2 in NPC and adjacent tissues and in different NPC cell lines and a normal nasal mucosal epithelial cell line (HNEpC). Experiments of PRMT1 or RRM2 overexpression or siRNA-mediated PRMT1 or RRM2 knockdown were carried out in CNE-2 cells to investigate the relationship between PRMT1 and RRM2 expressions using Western blotting. Apoptosis of the transfected cells was detected using Annexin V-FITC/PI apoptosis detection kit, and the production of intracellular reactive oxygen species (ROS) was determined using a ROS detection kit.

RESULTS

Compared with adjacent tissues and HNEpC cells, NPC tissues and cell lines expressed significantly higher levels of PRMT1 and RRM2 ( < 0.05). In CNE-2 cells with PRMT1 or RRM2 overexpression or knockdown, Western blotting demonstrated that PRMT1 could positively regulate the expression of RRM2 ( < 0.05). Overexpression of PRMT1 or RRM2 significantly reduced intracellular ROS production and apoptosis rate of CNE-2 cells ( < 0.05), and PRMT1 or RRM2 knockdown strongly increased ROS production and cell apoptosis ( < 0.05). Overexpression of either PRMT1 or RRM2 significantly decreased the expressions of cleaved caspase-3 and cleaved caspase-8 proteins ( < 0.05), and PRMT1 or RRM2 knockdown obviously promoted their expressions ( < 0.05). PRMT1 knockdown combined with RRM2 overexpression, as compared with PRMT1 knockdown only, significantly decreased ROS production and cell apoptosis ( < 0.05) as well as the protein expressions of cleaved caspase-3 and cleaved caspase-8 in CNE-2 cells ( < 0.05).

CONCLUSION

The high expression of PRMT1 in NPC inhibits apoptosis of NPC cells by promoting the expression of RRM2.

摘要

目的

验证蛋白精氨酸甲基转移酶1(PRMT1)是否通过促进核糖核苷酸还原酶M2(RRM2)的表达来抑制鼻咽癌(NPC)细胞的凋亡。

方法

采用免疫组织化学和蛋白质印迹法检测PRMT1和RRM2在NPC组织、癌旁组织、不同NPC细胞系及正常鼻黏膜上皮细胞系(HNEpC)中的相对表达。在CNE-2细胞中进行PRMT1或RRM2过表达实验,或采用小干扰RNA(siRNA)介导的PRMT1或RRM2敲低实验,运用蛋白质印迹法研究PRMT1与RRM2表达之间的关系。使用膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V-FITC/PI)凋亡检测试剂盒检测转染细胞的凋亡情况,采用活性氧(ROS)检测试剂盒测定细胞内ROS的产生。

结果

与癌旁组织和HNEpC细胞相比,NPC组织和细胞系中PRMT1和RRM2的表达水平显著更高(<0.05)。在PRMT1或RRM2过表达或敲低的CNE-2细胞中,蛋白质印迹法显示PRMT1可正向调控RRM2的表达(<0.05)。PRMT1或RRM2过表达显著降低了CNE-2细胞内ROS的产生及凋亡率(<0.05),而PRMT1或RRM2敲低则显著增加了ROS的产生及细胞凋亡(<0.05)。PRMT1或RRM2过表达均显著降低了裂解型半胱天冬酶-3和裂解型半胱天冬酶-8蛋白的表达(<0.05),PRMT1或RRM2敲低则明显促进了它们的表达(<0.05)。与单纯PRMT1敲低相比,PRMT1敲低联合RRM2过表达显著降低了CNE-2细胞内ROS的产生、细胞凋亡以及裂解型半胱天冬酶-3和裂解型半胱天冬酶-8的蛋白表达(<0.05)。

结论

NPC中PRMT1的高表达通过促进RRM2的表达来抑制NPC细胞的凋亡。

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