miR-324-5p通过调控KLF3/JAK2/STAT3通路对胰腺癌细胞耐药性的影响  

Effect of miR-324-5p on drug resistance of pancreatic cancer cells through regulating KLF3/JAK2/STAT3pathway

作　　者：陈小丽 田霞[1] 谭洁[1] 陈巍[1] 占婷 黄晓东[1] CHEN Xiaoli;TIAN Xia;TAN Jie;CHEN Wei;ZHAN Ting;HUANG Xiaodong(Department of Gastro-enterology,Wuhan Third Hospital/Tongren Hospital of Wuhan University,Hubei Wuhan 430060,China)

机构地区：[1]武汉大学附属同仁医院消化内科,湖北武汉430060

出　　处：《中国医院药学杂志》2024年第12期1371-1376,共6页Chinese Journal of Hospital Pharmacy

基　　金：武汉市卫健委青年项目(编号:WX20Q17);湖北省卫健委面上项目(编号:WJ2023M129)。

摘　　要：目的:探讨miR-324-5p对胰腺癌细胞耐药性的影响及其可能的分子机制,为寻找胰腺癌新的治疗靶点提供理论及实验依据。方法:收集化疗前后胰腺癌患者的血浆标本,用不同浓度的吉西他滨处理胰腺癌细胞,qRT-PCR检测吉西他滨处理后胰腺癌细胞及胰腺癌患者血浆miR-324-5p的表达。转染miR-324-5p模拟物或抑制剂,用一定浓度的吉西他滨处理胰腺癌细胞,流式细胞术检测细胞凋亡,Western blot检测凋亡相关蛋白的表达,观察miR-324-5p对细胞凋亡的影响。CCK-8法检测胰腺癌细胞吉西他滨的IC50,Western blot检测耐药相关蛋白的表达,观察miR-324-5p对细胞耐药的影响。转染KLF3过表达质粒,Western blot检测JAK2/STAT3信号通路的表达,进一步探讨miR-324-5p参与细胞耐药的分子机制。结果:miR-324-5p在吉西他滨处理的胰腺癌细胞及接受化疗的胰腺癌患者血浆中高表达。miR-324-5p可抑制吉西他滨处理后胰腺癌细胞凋亡及凋亡相关蛋白的表达。miR-324-5p可增强胰腺癌细胞对吉西他滨的耐药性及耐药相关蛋白的表达。过表达KLF3不仅可以部分逆转miR-324-5p对细胞凋亡及凋亡相关蛋白表达的影响,也可部分逆转miR-324-5p对细胞耐药及耐药相关蛋白表达的影响,这可能是通过调控JAK2/STAT3信号通路实现的。结论:miR-324-5p与胰腺癌细胞耐药相关,并可通过调控KLF3激活JAK2/STAT3信号通路影响胰腺癌细胞耐药。OBJECTIVE To explore the effect of miR-324-5p on drug resistance of pancreatic cancer(PC)cells,elucidate its potential molecular mechanism and provide theoretical and experimental rationales for seeking new therapeutic targets for PC.METHODS Plasma samples were collected from PC patients before and after chemotherapy.And PC cells were treated with different concentrations of gemcitabine(Gem).Quantitative real-time polymerase chain reaction(qRT-PCR)was employed for detecting the expression of miR-324-5p in PC cells and plasma of PC patients.Cells were transfected with miR-324-5p mimics or inhibitors and treated with a certain concentration of Gem.Flow cytometry was utilized for detecting cell apoptosis and Western blot for assessing the expressions of apoptosis-related proteins and examining the effect of miR-324-5p on cell apoptosis.IC50 of Gem in cells was detected by CCK-8.And Western blot was utilized for detecting the expressions of drug resistance-related proteins and observing the effect of miR-324-5p on drug resistance.To further explore the molecular mechanism of miR-324-5p involved in drug resistance,Western blot was utilized for detecting the expression of JAK2/STAT3 signaling pathway after transfecting KLF3 overexpression plasmid.RESULTS miR-324-5p was raised in PC cells treated with Gem and plasma of PC patients on chemotherapy.And miR-324-5p suppressed cell apoptosis and apoptosis-related protein expression in PC cells treated with Gem.miR-324-5p boosted cell drug resistance to Gem and enhanced the expressions of drug resistance-related proteins.KLF3 overexpression not only partially reversed the effects of miR-324-5p on cell apoptosis and apoptosis-related protein expression,but also partially reversed the effects of miR-324-5p on drug resistance and drug resistance-related protein expression in cells.Both effects were achieved through regulating the signaling pathway of JAK2/STAT3.CONCLUSION miR-324-5p is correlated with drug resistance of PC cells through activating JAK2/STAT3 signal via KLF3.

关 键 词：胰腺癌 miR-324-5p 耐药性 JAK2/STAT3通路 吉西他滨 

分 类 号：R966[医药卫生—药理学]