【摘要】 目的:探究环状RNA ciRS-7与子宫内膜癌(EC)细胞内质网(ER)应激之间的关系。方法:通过慢病毒转染细胞的方法构建稳定过表达ciRS-7的EC细胞系Ishikawa,集落形成试验检测过表达ciRS-7的Ishikawa细胞增殖活性变化，划痕试验检测Ishikawa细胞迁移能力变化，Transwell试验检测Ishikawa细胞侵袭能力变化，qRT-PCR检测Ishikawa细胞内miR-7表达水平变化。对Ishikawa细胞进行血清饥饿24h处理，通过Western blot检测Ishikawa细胞ER应激相关蛋白和凋亡相关蛋白表达水平，qRT-PCR检测Ishikawa细胞内miR-7表达水平。通过细胞免疫荧光试验比较饥饿处理的ciRS-7过表达细胞和和未经饥饿处理的空白组细胞内Ki-67和抗凋亡蛋白表达水平。构建稳定过表达ciRS-7和miR-7的Ishikawa细胞株，在饥饿处理细胞诱导ER应激后，Western blot检测ER应激相关蛋白和促凋亡蛋白表达水平。构建稳定敲低ciRS-7的Ishikawa细胞株，在饥饿处理细胞诱导ER应激后，通过qRT-PCR检测细胞miR-7水平，Western blot检测ER应激相关蛋白和促凋亡蛋白表达水平。结果:过表达ciRS-7的Ishikawa细胞增殖活性显著提升，迁移和侵袭能力显著增强；ciRS-7的过表达导致miR-7水平显著降低。血清饥饿24h成功诱导Ishikawa细胞ER应激水平升高。过表达ciRS-7的Ishikawa细胞内miR-7水平显著低于阴性对照组细胞，对ER应激抗性更强，促凋亡蛋白表达水平显著低于阴性对照组细胞。CiRS-7和miR-7双重过表达细胞对饥饿诱导的ER应激抗性降低，凋亡蛋白表达水平显著升高。敲低ciRS-7导致细胞miR-7水平升高，细胞对饥饿诱导的ER应激抗性减弱，细胞凋亡增加。结论:ciRS-7的表达能够促进EC细胞增殖、迁移和侵袭，并且通过抑制miR-7的表达增强EC细胞对饥饿诱导的ER应激抗性。更多还原

【Abstract】 Objective: To explore the relationship between circular RNA CiRS-7 and endoplasmic reticulum(ER) stress in endometrial carcinoma(EC) cells.Methods: An EC cell line, Ishikawa, was stably transfected with ciRS-7 using lentiviral vectors. Ishikawa cells’ proliferation activity, scratch wound healing, migration ability, and invasion capability were assessed. QRT-PCR determined miR-7 expression levels in these cells. Ishikawa cells were serum-starved for 24 hours, and the expression levels of ER stress-related and apoptosis-related proteins were measured by Western blot, with miR-7 expression quantified by qRT-PCR. Intracellular levels of Ki-67 and anti-apoptotic protein expression were compared between serum-starved ciRS-7-overexpressing cells and blank control groups. Ishikawa cell lines stably overexpressing both ciRS-7 and miR-7 were constructed, and Western blot was used to measure the levels of ER stress-related proteins and pro-apoptotic protein expression after ER stress induction. An Ishikawa cell line with stable ciRS-7 knockdown was established, and after ER stress induction in serum-starved cells, miR-7 levels were detected by qRT-PCR, while the expression levels of ER stress-related and pro-apoptotic proteins were assessed by Western blot.Results: Ishikawa cells overexpressing ciRS-7 exhibited significantly increased proliferative activity, migration, and invasion. Overexpression of ciRS-7 led to a significant decrease in miR-7 levels. Serum starvation for 24 hours successfully induced increased ER stress levels in Ishikawa cells. Intracellular miR-7 levels in ciRS-7-overexpressing Ishikawa cells were significantly lower than those in negative control cells, rendering them more resistant to ER stress and exhibiting significantly lower levels of pro-apoptotic proteins. Dual overexpression of ciRS-7 and miR-7 resulted in decreased resistance to starvation-induced ER stress and significantly higher expression levels of apoptotic proteins. Knockdown of ciRS-7 increased cellular miR-7 levels, diminished cell resistance to starvation-induced ER stress, and promoted apoptosis.Conclusion: CiRS-7 expression can promote EC cell proliferation, migration, and invasion, and enhance EC cell resistance to starvation-induced ER stress by inhibiting miR-7 expression.更多还原