lncRNA H19靶向miR-4735-3p激活NF-κB信号通路调控宫颈癌顺铂耐药  

lncRNA H19 Activates NF-κB Signaling Pathway by Targeting miR-4735-3p to Regulate Cisplatin Resistance in Cervical Cancer

作　　者：王烈宏[1] 祁青玲[1] 王乾印 高文君 王爱敏 曾慧 Wang Liehong;Qi Qingling;Wang Qianyin(Department of Gynecology,Qinghai Red Cross Hospital,Xining 810099,China)

机构地区：[1]青海红十字医院妇科,西宁810099

出　　处：《华中科技大学学报（医学版）》2023年第4期496-502,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：青海省科技计划项目(No.2021-ZJ-764)。

摘　　要：目的揭示lncRNA H19调控宫颈癌细胞顺铂(DDP)耐药的作用及分子机制。方法体外建立顺铂耐药的宫颈癌细胞株HeLa/DDP。qRT-PCR检测H19和miR-4735-3p的表达。CCK-8实验检测不同浓度DDP作用下细胞的增殖抑制率,计算IC50值。克隆形成实验评估细胞增殖能力。流式细胞术检测细胞凋亡率。Western blot实验检测凋亡相关蛋白Bcl-2、Bax和c-Caspase3(c-Casp3)及NF-κB信号通路蛋白(p-IκBα/IκBα和p-p65/p65)的表达。双荧光素酶报告基因实验和RNA交互沉降实验验证H19和miR-4735-3p之间的靶向关系。结果H19在HeLa/DDP细胞中高表达。敲低H19增强HeLa/DDP对顺铂的敏感性,促进细胞凋亡。H19能够与miR-4735-3p靶向结合,敲低H19上调HeLa/DDP细胞中miR-4735-3p的表达。过表达miR-4735-3p增强HeLa/DDP对顺铂的敏感性,促进细胞凋亡。抑制miR-4735-3p能够逆转H19敲低对HeLa/DDP顺铂敏感性的增强作用。敲低H19通过靶向miR-4735-3p抑制p-IκBα和p-p65蛋白表达。结论H19通过靶向miR-4735-3p,抑制NF-κB通路活性,增强HeLa/DDP细胞的顺铂敏感性。Objective To reveal the role and molecular mechanism of lncRNA H19in regulating cisplatin resistance in cervical cancer cells.Methods Cisplatin-resistant cervical cancer cell line HeLa/DDP cells were established in vitro.The inhibition rate of cell proliferation under different concentrations of DDP was detected by CCK-8,and IC50value was calculated.Cell proliferative capacity was assessed by colony formation assay.Cell apoptosis rate was detected by flow cytometry.Western blot assay was used to detect the expressions of apoptosis-related proteins B-cell lymphoma-2(Bcl-2),Bcl-2related X protein(Bax),c-Caspase3(c-Casp3),and NF-κB signaling pathway proteins(p-IκBα/IκBαand p-p65/p65).Results lncRNA H19was highly expressed in HeLa/DDP cells.Knockdown of H19enhanced the sensitivity of HeLa/DDP to cisplatin and promoted apoptosis.H19could target miR-4735-3p,knockdown of H19 upregulated the expression of miR-4735-3p in HeLa/DDP cells.Overexpression of miR-4735-3p enhanced the sensitivity of HeLa/DDP to cisplatin and promoted apoptosis.Inhibition of miR-4735-3p reversed the promoting effects of H19knockdown on HeLa/DDP cisplatin sensitivity.Knockdown of H19inhibited p-IκBαand p-p65protein expression by targeting miR-4735-3p.Conclusion H19enhanced cisplatin sensitivity in HeLa/DDP cells by targeting miR-4735-3p and inhibiting NF-κB pathway activity.

关 键 词：宫颈癌 顺铂 lncRNA H19 miR-4735-3p NF-ΚB信号通路 

分 类 号：R737.33[医药卫生—肿瘤]