miR-548o-3p负反馈失调致宫颈癌紫杉醇抗性的机制  

Mechanism of miR-548o-3p negative feedback imbalance leading to paclitaxel resistance in cervical cancer

作　　者：陈娜[1] 崔建涛 高娜[1] 张玉丽 陈秀英[1] 李晓丹 卞欣 张士表[1] CHEN Na;CUI Jiantao;GAO Na;ZHANG Yuli;CHEN Xiuying;LI Xiaodan;BIAN Xin;ZHANG Shibiao(Department of Gynecology,Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine,Cangzhou 061013,China)

机构地区：[1]河北省沧州中西医结合医院妇一科,河北沧州061013

出　　处：《生物技术》2024年第5期582-588,547,共8页Biotechnology

基　　金：2023年度河北省医学科学研究课题(20232140)。

摘　　要：[目的]探讨宫颈癌紫杉醇抗性形成的潜在机制。[方法]紫杉醇浓度从0.001μmol/L逐渐递增至0.1μmol/L处理Hela细胞超过12个月进行Hela紫杉醇抗性细胞(Hela/TR)的筛选。检测Hela与Hela/TR细胞的细胞活力、凋亡水平、细胞周期和肿瘤干细胞水平。紫杉醇处理后,检测Hela与Hela/TR细胞内紫杉醇浓度。[结果]紫杉醇处理可以显著抑制Hela细胞的增殖并且诱导细胞凋亡。与Hela细胞相比,Hela/TR细胞中CD44^(+)细胞的比例显著增加,并且Hela/TR细胞中肿瘤干细胞标志物SOX2和ALDH1的表达水平高于Hela细胞。与Hela细胞相比,Hela/TR细胞的细胞内紫杉醇浓度明显降低(0.0759±0.0130 vs 0.0031±0.0004,t=12.52,P<0.05)。与Hela细胞相比,Hela/TR细胞中ABCC5的表达水平显著上升(0.15±0.04 vs 0.72±0.04,t=22.53,P<0.05)。敲低ABCC5后,Hela/TR细胞的细胞内紫杉醇浓度明显升高。与Hela细胞相比,Hela/TR细胞中FOXM1的表达水平上升。敲低FOXM1后,Hela/TR细胞中ABCC5的表达水平下降(0.13±0.07 vs 0.64±0.03,t=14.97,P<0.05),细胞内紫杉醇浓度明显上升。过表达miR-548o-3p后,Hela/TR细胞中FOXM1和ABCC5的水平表达下降,细胞内紫杉醇浓度明显上升(0.003±0.000 vs 0.072±0.010,t=15.43,P<0.05)。miR-548o-3p靶向FOXM1 mRNA的3′端非翻译区。过表达miR-548o-3p后,Hela/TR细胞对紫杉醇的抵抗显著下降(0.51±0.05 vs 0.10±0.01,t=17.98,P<0.05)。[结论]宫颈癌紫杉醇抗性细胞中miR-548o-3p负反馈失调导致FOXM1/ABCC5轴过度激活,提升了紫杉醇从宫颈癌细胞内的排除效率,增强了宫颈癌细胞的活力。[Objective]To explore the potential mechanism of paclitaxel resistance in cervical cancer.[Method]The paclitaxel concentration was increased from 0.001μmol/L to 0.1μmol/L to treat Hela cells for over 12 months for the selection of Hela-paclitaxel-resistant cells(Hela/TR).The cell viability,apoptosis level,cell cycle and tumor stem cell level of Hela and Hela/TR cells were detected.After paclitaxel treatment,the intracellular paclitaxel concentration in Hela and Hela/TR cells was detected.[Result]Paclitaxel treatment significantly inhibited the proliferation and induced apoptosis of Hela cells.The proportion of CD44^(+)cells was significantly increased in Hela/TR cells compared with Hela cells,and the expression levels of cancer stem cell markers SOX2 and ALDH1 were much higher in Hela/TR cells than in Hela cells.The intracellular paclitaxel concentration of Hela/TR cells was significantly lower than that of Hela cells(0.0759±0.0130 vs 0.0031±0.0004,t=12.52,P<0.05).Compared with Hela cells,the expression level of ABCC5 in Hela/TR cells was significantly increased(0.15±0.04 vs 0.72±0.04,t=22.53,P<0.05).After knockdown of ABCC5,the intracellular concentration of paclitaxel in Hela/TR cells was significantly increased.The expression level of FOXM1 was increased in Hela/TR cells compared with Hela cells.After knockdown of FOXM1,the expression level of ABCC5 in Hela/TR cells decreased(0.13±0.07 vs 0.64±0.03,t=14.97,P<0.05),and the intracellular concentration of paclitaxel increased significantly.After over-expression of miR-548o-3p,the expression levels of FOXM1 and ABCC5 in Hela/TR cells were decreased,and the intracellular concentration of paclitaxel was significantly increased(0.003±0.000 vs 0.072±0.010,t=15.43,P<0.05).miR-548o-3p targeted the 3′untranslated region of FOXM1 mRNA.After overexpression of miR-548o-3p,the resistance of Hela/TR cells to paclitaxel was significantly decreased(0.51±0.05 vs 0.10±0.01,t=17.98,P<0.05).[Conclusion]The negative feedback dysregulation of miR-548o-3p in cervical can

关 键 词：miR-548o-3p FOXM1 ABCC5 宫颈癌 紫杉醇 耐药 HELA 凋亡 

分 类 号：R737.3[医药卫生—肿瘤]