转铁蛋白受体对宫颈癌细胞自噬的影响

Effect of Transferrin Receptor on Autophagy of Cervical Cancer Cells

目的:探讨敲低转铁蛋白受体(TFRC)对宫颈癌Hela细胞增殖、迁移和自噬的影响及可能的作用机制。方法:慢病毒构建敲低TFRC宫颈癌细胞系,分为空白对照组、TFRC敲低阴性对照组、TFRC敲低组。通过蛋白质印迹验证各组目的基因TFRC、自噬相关蛋白Beclin-1、P62、LC3和细胞外信号调节激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(ERK/AKT/mTOR)信号通路相关蛋白的表达水平。利用CCK8法检测各组细胞增殖活性;流式细胞术检测各组细胞周期分布;细胞划痕实验检测各组细胞迁移能力。结果:与空白对照组相比,TFRC敲低组自噬相关蛋白Beclin-1、LC3蛋白表达水平降低(P P P P P P < 0.01)。结论:敲低转铁蛋白受体(TFRC)可能通过抑制ERK/AKT/mTOR信号通路抑制宫颈癌细胞自噬,进而抑制其增殖、迁移。

Objective: To investigate the effects of knockdown of transferrin receptor on proliferation, migration and autophagy of Hela cells of cervical cancer, and to study of possible molecular mechanisms. Methods: Lentiviral construction of knockdown TFRC cervical cancer cell lines, and were divided into the blank control group, TFRC knock-down negative control group, TFRC knock-down group. The expression of target gene TFRC, autophagy-related proteins Beclin-1, P62, LC3, and extracellular signal-regulated kinase/protein kinase B/mammalian target of rapamycin (ERK/AKT/mTOR) signaling pathway-related proteins was detected by Western blotting. The activities of cells in various group were detected by CCK-8 assay. Cell cycle distributions in various group were detected by flow cytometry. Cell migration abilities in various group were detected by cell scratch assay. Observation cellular autophagic vesicles in various group by electron microscopy. Results: Compared with the blank control group, the protein expression levels of autophagy-related proteins Beclin-1 and LC3 in cervical cancer cells in TFRC knock-down group decreased (P P P P P P < 0.01). Conclusions: Knockdown of transferrin receptor (TFRC) may inhibit autophagy of cervical cancer cells by suppressing the ERK/AKT/mTOR signaling pathway, which in turn inhibits their proliferation and migration.