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。
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.
作者
安欣
崔志利
魏娉
杨学丽
江现丽
何鹏
AN Xin(The First Hospital of Handan,Hebei Handan 056002,China)
出处
《河北医学》
2025年第1期52-56,共5页
Hebei Medicine
基金
河北省2022年度医学科学研究课题,(编号:20220502)。
