miR-17-5p靶向PTEN促进非小细胞肺癌吉非替尼耐药
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摘要
目的探讨miR-17-5p在NSCLC吉非替尼耐药中的作用。方法 QRT-PCR法检测miR-17-5p在细胞模型中的表达。MTS法检测细胞过表达或敲低miR-17-5p后的耐药性变化。预测miR-17-5p下游靶基因并通过双荧光素酶报告基因实验、功能阻断实验等证实miR-17-5p通过靶向PTEN基因介导对吉非替尼的耐药。QRT-PCR法检测耐药前后NSCLC患者血清中的miR-17-5p表达。结果 miR-17-5p在耐药细胞中表达增加(P <0.05)。敏感与耐药细胞中分别过表达和敲低miR-17-5p后,细胞对吉非替尼耐药性分别增加和降低(P <0.05)。发现PTEN为miR-17-5p下游靶基因,敲低PTEN后可逆转敲低miR-17-5p导致的耐药性降低。耐药后NSCLC患者血清中miR-17-5p表达升高。结论 miR-17-5p通过下调PTEN表达促进了NSCLC对吉非替尼的耐药。
Objective To investigate the role of miR-17-5 p in non-small cell lung cancer(NSCLC)gefitinib resistance. Methods The expression of miR-17-5 p in cell models was detected by QRT-PCR. MTS assay was used to detect changes in drug resistance after over-expressing or knocking down miR-17-5 p in cells. The miR-17-5 p downstream target gene was predicted and miR-17-5 p mediated resistance to gefitinib by targeting target genes was confirmed by double luciferase reporter gene experiment,functional blocking experiment,etc. The expression of miR-17-5 p in serum of NSCLC patients before and after drug resistance was detected by QRT-PCR. Results The expression of miR-17-5 p in drug-resistant cells increased(P < 0.05). Gefitinib resistance increased and decreased after over-expression and knockdown of miR-17-5 p in sensitive and resistant cells,respectively(P < 0.05). PTEN was found to be a downstream target gene of miR-17-5 p,and knockdown of PTEN reversed the decrease in drug resistance caused by knockdown of miR-17-5 p. The expression of miR-17-5 p in serum of NSCLC patients increased after drug resistance. Conclusion miR-17-5 p promotes gefitinib resistance in NSCLC by down-regulating PTEN gene expression.
Objective To investigate the role of miR-17-5 p in non-small cell lung cancer(NSCLC)gefitinib resistance. Methods The expression of miR-17-5 p in cell models was detected by QRT-PCR. MTS assay was used to detect changes in drug resistance after over-expressing or knocking down miR-17-5 p in cells. The miR-17-5 p downstream target gene was predicted and miR-17-5 p mediated resistance to gefitinib by targeting target genes was confirmed by double luciferase reporter gene experiment,functional blocking experiment,etc. The expression of miR-17-5 p in serum of NSCLC patients before and after drug resistance was detected by QRT-PCR. Results The expression of miR-17-5 p in drug-resistant cells increased(P < 0.05). Gefitinib resistance increased and decreased after over-expression and knockdown of miR-17-5 p in sensitive and resistant cells,respectively(P < 0.05). PTEN was found to be a downstream target gene of miR-17-5 p,and knockdown of PTEN reversed the decrease in drug resistance caused by knockdown of miR-17-5 p. The expression of miR-17-5 p in serum of NSCLC patients increased after drug resistance. Conclusion miR-17-5 p promotes gefitinib resistance in NSCLC by down-regulating PTEN gene expression.
引文
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[2]O′KANE G M,BAMES T A,LEIGH N B.Resistance to epidermal growth factor receptor tyrosine kinase inhibitors,T790M,and clinical trials[J].Curr Oncol,2018,25(1):28-37.
[3]MCGUIRE A,BROWN J A,KERIN M J.Metastatic breast cancer:the potential of miRNA for diagnosis and treatment monitoring[J].Cancer Metastasis Rev,2015,34(1):145-155.
[4]SHIN V Y,CHU K M.MiRNA as potential biomarkers and therapeutic targets for gastric cancer[J].World J Gastroenterol,2014,20(30):10432-10439.
[5]罗凯.PFTK1在非小细胞肺癌EGFR-TKIs耐药中的作用及机制[D].广州医科大学,2017.
[6]吴莹莹,高波,丁跃明,等.miR-17-5p在肿瘤中作用的研究进展[J].中国肿瘤临床,2016,43(3):130-134.
[7]WU Y L,ZHOU C,LIAM C K,et al.First-line erlotinib versus gemcitabine/cisplatin in patients with advanced EGFR mutation-positive non-small-cell lung cancer:Analyses from the phase III,randomized,open-label,ENSURE study[J].Ann Oncol,2015,26(9):1883-1889.
[8]MORGILLO F,DELLA CORTE C M,FASANO M,et al.Mechanisms of resistance to EGFR-targeted drugs:Lung cancer.[J].Esmo Open,2016,1(3):e000060.
[9]FUZIWARA C S,KIMURA E T.Insights into regulation of the miR-17-92 cluster of miRNAs in cancer[J].Front Med,2015,2(64):64.
[10]CHEN C,LU Z,YANG J,et al.MiR-17-5p promotes cancer cell proliferation and tumorigenesis in nasopharyngeal carcinoma by targeting p21[J].Cancer Med,2016,5(12):3489-3499.
[11]宋先璐,廖志伟,余宏伟,等.mir-17-5p、mir-92a、let-7b表达水平与非小细胞肺癌顺铂耐药的关系[J].分子影像学杂志,2016,39(3):286-291.
[12]BOBBILI M R,MADER R M,GRILLARI J,et al.OncomiR-17-5p:Alarm signal in cancer?[J].Oncotarget,2017,8(41):71206.
[13]SUN Y,TIAN H,WANG L.Effects of PTEN on the proliferation and apoptosis of colorectal cancer cells via the phosphoinositol-3-kinase/Akt pathway[J].Oncol Rep,2015,33(4):1828-1836.