CDCA4通过调节自噬抑制非小细胞肺癌中的上皮-间质转化（EMT）和转移。
CDCA4 suppresses epithelial-mesenchymal transtion (EMT) and metastasis in Non-small cell lung cancer through modulating autophagy.
作者信息
Xu Chenxin, Cao Haixia, Sui Ying, Zhang Hui, Shi Chen, Wu Jianzhong, Ma Rong, Feng Jifeng
机构信息
The Affiliated Cancer Hospital of Nanjing Medical University, Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research, No.42, Baiziting Street, Nanjing, 210009, Jiangsu, China.
Research Center for Clinical Oncology, Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research & The Affiliated Cancer Hospital of Nanjing Medical University, Nanjing, 210009, Jiangsu, China.
出版信息
Cancer Cell Int. 2021 Jan 12;21(1):48. doi: 10.1186/s12935-021-01754-w.
BACKGROUND
Cell division cycle associated 4 (CDCA4) has been reported to be engaged into the progression of several cancers. The function of CDCA4 in Non-small cell lung cancer (NSCLC) was unknown. We aimed to explore the critical role of CDCA4 in NSCLC.
METHODS
CDCA4 stably knocking down and overexpression cell lines were established and Western blotting assay was applied to measure relevant protein expression of Epithelial-Mesenchymal Transtion (EMT) and cell autophagy. Staining of acidic vacuoles, transmission electron microscopy and immunofluorescence staining were employed to detect autophagy. The ability of cells to migrate and invade were detected by Transwell migration and invasion assays. The interaction of CDCA4 with CARM1 was identified by immunoprecipitation and Western blotting analysis.
RESULTS
In the present study, it was found that inhibition of CDCA4 induced EMT, migration and invasion of NSCLC cells while inhibiting autophagy of NSCLC cells. Meanwhile, overexpression of CDCA4 in NSCLC cells showed the opposite function. More importantly, the inhibition of autophagy could promote the EMT, migration and invasion of NSCLC cells, which should be impaired via the activation of autophagy. In addition, CDCA4-inhibited EMT, migration and invasion could be partially aggravated by autophagy activator, rapamycin, and reversed by autophagy inhibitor, 3-MA. Correspondingly, the application of rapamycin or 3-MA to CDCA4 knockdown cells showed the opposite effects. Further investigation suggested that CDCA4 could interact with coactivator associated arginine methyltransferase 1 (CARM1). Autophagy was induced while cell migration and invasion were inhibited in CARM1 knockdown cells. CDCA4 could suppress the protein expression CARM1 and knocking down of CARM1 could alter cell autophagy, migratory and invasive abilities regulated by CDCA4.
CONCLUSION
All data indicated that CDCA4 inhibited the EMT, migration and invasion of NSCLC via interacting with CARM1 to modulate autophagy.
背景
据报道，细胞分裂周期相关蛋白4（CDCA4）参与多种癌症的进展。CDCA4在非小细胞肺癌（NSCLC）中的功能尚不清楚。我们旨在探讨CDCA4在NSCLC中的关键作用。
方法
建立CDCA4稳定敲低和过表达细胞系，采用蛋白质免疫印迹法检测上皮-间质转化（EMT）和细胞自噬的相关蛋白表达。采用酸性空泡染色、透射电子显微镜和免疫荧光染色检测自噬。通过Transwell迁移和侵袭试验检测细胞的迁移和侵袭能力。通过免疫沉淀和蛋白质免疫印迹分析鉴定CDCA4与共激活因子相关精氨酸甲基转移酶1（CARM1）的相互作用。
结果
在本研究中，发现抑制CDCA4可诱导NSCLC细胞发生EMT、迁移和侵袭，同时抑制NSCLC细胞的自噬。同时，NSCLC细胞中CDCA4的过表达显示出相反的功能。更重要的是，抑制自噬可促进NSCLC细胞的EMT、迁移和侵袭，而自噬的激活可削弱这些作用。此外，自噬激活剂雷帕霉素可部分加重CDCA4抑制的EMT、迁移和侵袭，自噬抑制剂3-甲基腺嘌呤（3-MA）可逆转这些作用。相应地，将雷帕霉素或3-MA应用于CDCA4敲低细胞显示出相反的效果。进一步研究表明，CDCA4可与共激活因子相关精氨酸甲基转移酶1（CARM1）相互作用。CARM1敲低细胞中诱导了自噬，同时抑制了细胞迁移和侵袭。CDCA4可抑制CARM1的蛋白表达，敲低CARM1可改变CDCA4调节的细胞自噬、迁移和侵袭能力。
结论
所有数据表明，CDCA4通过与CARM1相互作用调节自噬，从而抑制NSCLC的EMT、迁移和侵袭。
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