摘要
目的:探讨雷公藤甲素通过微小核糖核酸-138-5p(miR-138-5p)/染色质重塑因子1(RSF-1)调控宫颈癌顺铂(DDP)耐药和血管生成的相关机制。方法:选用人宫颈癌细胞(HeLa)、人胚肾细胞(293T)、人脐静脉内皮细胞(HUVEC)进行体外实验,建立HeLa/DDP耐药细胞株,按简单随机法将HeLa/DDP分为顺铂+HeLa/DDP组、雷公藤甲素+顺铂+HeLa/DDP组、雷公藤甲素+顺铂+miR-138-5p抑制剂(miR-138-5p inhibitor)+HeLa/DDP组、雷公藤甲素+顺铂+miR-138-5p inhibitor+小干扰核酸染色质重塑因子1(si-RSF-1)+HeLa/DDP组,将293T分为空白对照模拟物(NC mimic)组、miR-138-5p mimic组,将HUVEC分为顺铂+HUVEC组、雷公藤甲素+顺铂+HUVEC组、雷公藤甲素+顺铂+miR-138-5p inhibitor+HUVEC组、雷公藤甲素+顺铂+miR-138-5p inhibitor+si-RSF-1+HUVEC组。检测各组细胞增殖率、迁移能力、miR-138-5p和RSF-1水平并验证RSF-1与miR-138-5p的靶向关系和对其细胞小管形成的影响。结果:与顺铂+HeLa/DDP组比较,雷公藤甲素+顺铂+HeLa/DDP组细胞24、48、72 h增殖率、迁移能力降低(均P<0.05);与顺铂+HeLa/DDP组比较,雷公藤甲素+顺铂+HeLa/DDP组中miR-138-5p水平明显升高,而RSF-1明显降低(均P<0.05);与雷公藤甲素+顺铂+HeLa/DDP组比较,雷公藤甲素+顺铂+miR-138-5p inhibitor+HeLa/DDP组miR-138-5p表达水平明显降低,而RSF-1明显升高(均P<0.05);与雷公藤甲素+顺铂+miR-138-5p inhibitor+HeLa/DDP组比较,雷公藤甲素+顺铂+miR-138-5p inhibitor+si-RSF-1+HeLa/DDP组miR-138-5p水平无明显差异(P>0.05),而RSF-1明显降低(P<0.05);与NC mimic组比较,miR-138-5p mimic组共转染RSF-1-野生型质粒(RSF-1-WT)萤光素酶活性降低;与顺铂+HUVEC组比较,雷公藤甲素+顺铂+HUVEC组管状结构生成数量明显降低;与雷公藤甲素+顺铂+HUVEC组比较,雷公藤甲素+顺铂+miR-138-5p inhibitor+HUVEC组管状结构生成数量明显降低;与雷公藤甲素+顺铂+miR-138-5p inhibitor+HUVEC组比较,雷公藤甲素+顺铂+miR-138-5p inhibitor+si
Objective:To explore the mechanism of triptolide in regulating cisplatin(DDP)resistance and angiogenesis in cervical cancer through microRNA-138-5p(miR-138-5p)and chromatin remodeling factor 1(RSF-1).Methods:In vitro experiments were conducted using human cervical cancer cells(HeLa),human embryonic kidney cells(293T),and human umbilical vein endothelial cells(HUVEC).HeLa/DDP-resistant cell lines were established.HeLa/DDP cells were randomly divided into cisplatin+HeLa/DDP group,triptolide+cisplatin+HeLa/DDP group,triptolide+cisplatin+miR-138-5p inhibitor+HeLa/DDP group,and triptolide+cisplatin+miR-138-5p inhibitor+si-RSF-1+HeLa/DDP group.The 293T cells were divided into NC mimic group and miR-138-5p mimic group.HUVECs were categorized into cisplatin+HUVEC group,triptolide+cisplatin+HUVEC group,triptolide+cisplatin+miR-138-5p inhibitor+HUVEC group,and triptolide+cisplatin+miR-138-5p inhibitor+si-RSF-1+HUVEC group.The proliferation rate,migration ability,levels of miR-138-5p and RSF-1 were measured,and the targeting relationship between RSF-1 and miR-138-5p,as well as its effect on tube formation,were verified.Results:Compared to the cisplatin+HeLa/DDP group,the proliferation rate and migration ability of the triptolide+cisplatin+HeLa/DDP group decreased at 24,48,and 72 h(all P<0.05).In the triptolide+cisplatin+HeLa/DDP group,miR-138-5p levels significantly increased while RSF-1 levels decreased(both P<0.05)compared to the cisplatin+HeLa/DDP group.In the triptolide+cisplatin+miR-138-5p inhibitor+HeLa/DDP group,miR-138-5p expression levels significantly decreased while RSF-1 levels increased(both P<0.05)compared to the triptolide+cisplatin+HeLa/DDP group.Compared with the triptolide+cisplatin+miR-138-5p inhibitor+HeLa/DDP group,the triptolide+cisplatin+miR-138-5p inhibitor+si-RSF-1+HeLa/DDP group showed no significant difference in miR-138-5p levels(P>0.05),but RSF-1 levels significantly decreased(P<0.05).Co-transfection of RSF-1-WT with miR-138-5p mimic resulted in reduced luciferase activity compared to the NC mim
作者
马爱迪
张玥
蒋奇欣
曲琰
张秋华
MA Aidi;ZHANG Yue;JIANG Qixin;QU Yan;ZHANG Qiuhua(Department of Pharmacy,Xinglin College of Liaoning University of Traditional Chinese Medicine,Shenyang 110000,China;Department of Traditional Chinese Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110000,China)
出处
《世界中医药》
CAS
北大核心
2024年第17期2572-2577,2583,共7页
World Chinese Medicine
基金
国家自然科学基金项目(81803726)
辽宁省中央引导地方科技发展专项项目(2023010303-JH6/1001)。
