Department of Medical Laboratory, West China Second University Hospital, Sichuan University, Chengdu, Sichuan Province, China; Key Laboratory of Obstetric & Gynecologic and Pediatric Diseases and Brith Defects of Ministry of Education, Chengdu, Sichuan Province, China.
Institute of Health Policy & Hospital Management, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chinese Academy of Sciences Sichuan Translational Medicine Research Hospital, Chengdu, Sichuan Province, China; West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu, Sichuan Province, China.
J Reprod Immunol. 2020 Jun;139:103120. doi: 10.1016/j.jri.2020.103120. Epub 2020 Mar 15.
Cervical cancer cell function is influence by ER. Therefore, in this study, ER stress senser-ATF6, was selected for detailed research in cervical cancer.
ATF6 mRNA was assessed through RT-qPCR assays. Cell transfection was to regulate ATF6 and thereafter the differential ATF6 cancer cells were divided into two groups for further functional assays. Cell viabilities were analyzed by CCK-8 and migration by Scratch. RT-qPCR examined cell death biomarkers Caspas-3 and Bcl-2. 4-PBA was utilized to inhibit ER stress. After that, ATF6, viability, migration and apoptotic proteins were scrutinized after ER inhibition. Proteins signifying EMT, autophagy and MAPK signaling pathway were checked by western bolt. Last, we inactivated the MAPK signaling to investigate into the changes in cell functions.
ATF6 presented higher expression in cervical cancer cells. Inhibited ATF6 could reduce cell viabilities and migration but promote apoptosis through suppressing Bcl-2 and increasing caspase-3. ER stress antagonist witnessed a drop in ATF6 expression, cell viability, migration and Bcl-2 but a rise in caspase-3 activation, suggesting apoptosis increase. Cell autophagy was hindered in CC cells. Knockdown of ATF6 promoted autophagy and restrained EMT and MAPK signaling pathway. Suppressed ERK1/2 obstructed cell viabilities, migration, EMT and autophagy but promoted apoptosis.
ATF6 might promote cell growth, migration, autophagy through ER stress and MAPK signaling in cervical cancer in vitro, indicating a potential regulatory gene in cervical cancer. However, in-depth researches are requested to enrich the knowledge of ATF6 in cervical cancer in vivo and in clinical in the future.
ER 对宫颈癌细胞功能有影响。因此，在本研究中，选择 ER 应激感受器 ATF6 进行详细研究。
通过 RT-qPCR 检测 ATF6 mRNA。细胞转染用于调节 ATF6，此后将差异 ATF6 癌细胞分为两组进行进一步的功能测定。通过 CCK-8 分析细胞活力，通过划痕实验分析细胞迁移。RT-qPCR 检测细胞死亡标志物 Caspas-3 和 Bcl-2。用 4-PBA 抑制 ER 应激。之后，在 ER 抑制后检测 ATF6、活力、迁移和凋亡蛋白。通过 Western bolt 检测 EMT、自噬和 MAPK 信号通路相关蛋白。最后，我们失活 MAPK 信号通路，以研究细胞功能的变化。
ATF6 在宫颈癌细胞中表达较高。抑制 ATF6 可降低细胞活力和迁移，但通过抑制 Bcl-2 和增加 caspase-3 促进凋亡。ER 应激拮抗剂观察到 ATF6 表达下降，细胞活力、迁移和 Bcl-2 下降，caspase-3 激活增加，提示凋亡增加。CC 细胞中的细胞自噬受到抑制。ATF6 的敲低促进自噬并抑制 EMT 和 MAPK 信号通路。抑制 ERK1/2 可阻止细胞活力、迁移、EMT 和自噬，但促进凋亡。
ATF6 可能通过 ER 应激和 MAPK 信号通路促进宫颈癌体外细胞生长、迁移、自噬，提示其可能是宫颈癌的潜在调控基因。然而，未来需要深入研究以丰富 ATF6 在宫颈癌体内和临床中的知识。
