miR-103靶向PTEN并激活PI3K/AKT通路促进肺癌细胞A549对达沙替尼耐药
|
摘要
目的:探讨miR-103靶向PTEN并激活PI3K/AKT信号通路促进肺癌细胞对达沙替尼(dasatinib,DASA)耐药的机制。方法:收集2014年4月至2018年1月昆明医科大学第一附属医院胸外科收治的资料完整的肺癌DASA耐药组织和不耐药组织各35例。采用q PCR实验检测miR-103在肺癌DASA耐药组织和细胞中的表达水平,同时,采用CCK-8、Transwell和Wb实验检测敲降miR-103对A549/DASA细胞增殖、迁移和上皮间质转化(EMT)的影响,双荧光素酶报告基因验证miR-103与PTEN的靶向关系。进一步采用CCK-8、Transwell和Wb实验检测miR-103通过PTEN-PI3K/AKT信号通路对A549/DASA细胞恶性生物学行为的影响。结果:miR-103在肺癌DASA耐药组织和A549/DASA细胞中均高表达(均P<0.01)。敲降miR-103可显著抑制A549/DASA细胞的增殖、迁移和EMT(P<0.05或P<0.01)。此外,双荧光素酶报告基因证实miR-103靶向作用PTEN并下调其表达水平(P<0.01)。进一步实验显示,过表达mi R-103通过靶向下调PTEN并激活PI3K/AKT信号通路进而显著促进A549/DASA细胞增殖、迁移和EMT(P<0.05或P<0.01),从而上调A549/DASA细胞对DASA的耐药性。结论:miR-103/PTEN/PI3K/AKT信号通路与肺癌DASA耐药性存在调控关系,敲降miR-103可逆转A549/DASA对DASA耐药。
Objective: To explore the mechanism of miR-103 targeting PTEN(gene of phosphate and tension homology deleted on chromsome ten) and activating PI3K/AKT signaling pathway to promote dasatinib(DASA) resistance in lung cancer cells. Methods:DASA-resistant tissues and non-resistant tissues(35 samples for each) from patients treated in Department of Thoracic Surgery, the First Affiliated Hospital of Kunming Medical University from April 2014 to January 2018 were collected for this study. Expression of miR-103 was detected in DASA-resistant tissues and cell lines of lung cancer by quantitative Real-time polymerase chain reaction(qPCR). The effect of miR-103 knock-down on the proliferation, invasion and epithelial mesenchymal transition(EMT) of A549/DASA cells were measured by CCK-8 assay, Transwell and Wb, respectively. Subsequently, the dual luciferase reporter gene assay was used to verify whether PTEN was a target gene of miR-103. CCK-8, Transwell and Wb assay were further used to investigate the effect of miR-103 on malignant biological behaviors of A549/DASA cells via regulating PTEN-PI3K/AKT signaling pathway. Results: miR-103 was highly expressed in DASA-resistant tissues and A549/DASA cells(P<0.01). Knockdown of miR-103 significantly inhibited the proliferation, invasion and EMT of A549/DASA cells(P<0.05 or P<0.01). Additionally, dual luciferase reporter gene assay confirmed that miR-103 directly targeted PTEN and down-regulated its expression(P<0.01). Mechanistically, over-expression of miR-103 targeted and down-regulated PTEN to promote cell viability, invasion and EMT via activating PI3K/AKT pathway(P<0.05 or P<0.01), and further up-regulated the DASA-resistance in A549/DASA cells. Conclusion: miR-103/PTEN/PI3K/AKT signaling pathway plays a certain role in regulating DASA resistance of lung cancer, and knockdown of miR-103 expression may reverse the resistance of A549/DASA cells to DASA.
Objective: To explore the mechanism of miR-103 targeting PTEN(gene of phosphate and tension homology deleted on chromsome ten) and activating PI3K/AKT signaling pathway to promote dasatinib(DASA) resistance in lung cancer cells. Methods:DASA-resistant tissues and non-resistant tissues(35 samples for each) from patients treated in Department of Thoracic Surgery, the First Affiliated Hospital of Kunming Medical University from April 2014 to January 2018 were collected for this study. Expression of miR-103 was detected in DASA-resistant tissues and cell lines of lung cancer by quantitative Real-time polymerase chain reaction(qPCR). The effect of miR-103 knock-down on the proliferation, invasion and epithelial mesenchymal transition(EMT) of A549/DASA cells were measured by CCK-8 assay, Transwell and Wb, respectively. Subsequently, the dual luciferase reporter gene assay was used to verify whether PTEN was a target gene of miR-103. CCK-8, Transwell and Wb assay were further used to investigate the effect of miR-103 on malignant biological behaviors of A549/DASA cells via regulating PTEN-PI3K/AKT signaling pathway. Results: miR-103 was highly expressed in DASA-resistant tissues and A549/DASA cells(P<0.01). Knockdown of miR-103 significantly inhibited the proliferation, invasion and EMT of A549/DASA cells(P<0.05 or P<0.01). Additionally, dual luciferase reporter gene assay confirmed that miR-103 directly targeted PTEN and down-regulated its expression(P<0.01). Mechanistically, over-expression of miR-103 targeted and down-regulated PTEN to promote cell viability, invasion and EMT via activating PI3K/AKT pathway(P<0.05 or P<0.01), and further up-regulated the DASA-resistance in A549/DASA cells. Conclusion: miR-103/PTEN/PI3K/AKT signaling pathway plays a certain role in regulating DASA resistance of lung cancer, and knockdown of miR-103 expression may reverse the resistance of A549/DASA cells to DASA.
引文
[1] TORRE L A, BRAY F, SIEGEL R L, et al. Global cancer statistics,2012[J]. CA Cancer J Clin, 2015, 65(2):87-108. DOI:10.3322/caac.21262.
[2] WANG M, YUANG C, CHANG A. Molecular mechanism of action and potential biomarkers of growth inhibition of synergistic combination of afatinib and dasatinib against gefitinib-resistant non-small cell lung cancer cells[J]. Oncotarget, 2018, 9(23):16533-16546.DOI:10.18632/oncotarget.24814.
[3] BEAUCHAMP E M, WOODS B A, DULAK A M, et al. Acquired resistance to dasatinib in lung cancer cell lines conferred by DDR2gatekeeper mutation and NF1 loss[J]. Mol Cancer Ther, 2014, 13(2):475-482. DOI:10.1158/1535-7163.mct-13-0817.
[4] ZHANG Y, QU X, LI C, et al. miR-103/107 modulates multidrug resistance in human gastric carcinoma by downregulating Cav-1[J].Tumour Biol, 2015, 36(4):2277-2285. DOI:10.1007/s13277-014-2835-7.
[5] WAN L, TIAN Y, ZHANG R, et al. MicroRNA-103 confers the resistance to long-treatment of adriamycin to human leukemia cells by regulation of COP1[J]. J Cell Biochem, 2018, 119(5):3843-3852. DOI:10.1002/jcb.26431.
[6] CHEN H Y, LIN Y M, CHUANG H C, et al. miR-103/107 promote metastasis of colorectal cancer by targeting the metastasis suppressors DAPK and KLF4[J]. Cancer Res, 2017, 77(23):6788-6793.DOI:10.1158/0008-5472.can-17-3079.
[7] YANG D, WANG J J, LI J S, et al. miR-103 functions as a tumor suppressor by directly targeting programmed cell death 10 in NSCLC[J]. Oncol Res, 2017, 26(4):519-528 DOI:10.3727/096504017x15000757094686.
[8] ZHENG J, LIU Y, QIAO Y, et al. miR-103 promotes proliferation and metastasis by targeting KLF4 in gastric cancer[J/OL]. Int J Mol Sci, 2017, 18(5):pii:E910[2019-01-11]. https://www. ncbi. nlm. nih.gov/pubmed/28445396.DOI:10.3390/ijms18050910.
[9] PEREZ-RAMIREZ C, CANADAS-GARRE M, MOLINA M A, et al. PTEN and PI3K/AKT in non-small-cell lung cancer[J]. Pharmacogenomics, 2015, 16(16):1843-1862. DOI:10.2217/pgs.15.122.
[10] ZHANG R, LI G, ZHANG Q, et al. Hirsutine induces m PTP-dependent apoptosis through ROCK1/PTEN/PI3K/GSK3beta pathway in human lung cancer cells[J]. Cell Death Dis, 2018, 9(6):598-605.DOI:10.1038/s41419-018-0641-7.
[11] ZHU J, ZHU WWU W. MicroRNAs change the landscape of cancer resistance[J]. Methods Mol Biol, 2018, 1699(2):83-89. DOI:10.1007/978-1-4939-7435-1_6.
[12] MACDONAGH L, GRAY S G, FINN S P, et al. The emerging role of microRNAs in resistance to lung cancer treatments[J]. Cancer Treat Rev, 2015, 41(2):160-169. DOI:10.1016/j.ctrv.2014.12.009.
[13] WANG C, WANG S, MA F, et al. miRNA328 overexpression confers cisplatin resistance in nonsmall cell lung cancer via targeting of PTEN[J]. Mol Med Rep, 2018, 11(1):625-632 DOI:10.3892/mmr.2018.9478.
[14] SHEN H, ZHU F, LIU J, et al. Alteration in Mir-21/PTEN expression modulates gefitinib resistance in non-small cell lung cancer[J/OL]. PLoS One, 2014, 9(7):e103305[2019-01-11]. http://med.wanfangdata.com.cn/Paper/Detail/PeriodicalPaper_PM25058005.DOI:10.1371/journal.pone.0103305.
[15] WANG N, ZHU C, XU Y, et al. Negative regulation of PTEN by microRNA-221 and its association with drug resistance and cellular senescence in lung cancer cells[J]. Biomed Res Int, 2018, 2018:7908950. https://www. ncbi. nlm. nih. gov/pubmed/29876362. DOI:10.1155/2018/7908950.
[16] ZHANG Q, ZHANG B, SUN L, et al. MicroRNA-130b targets PTEN to induce resistance to cisplatin in lung cancer cells by activating Wnt/beta-catenin pathway[J]. Cell Biochem Funct, 2018, 36(4):194-202. DOI:10.1002/cbf.3331.
[17] FUMAROLA C, BONELLI M A, PETRONINI P G, et al. Targeting PI3K/AKT/mTOR pathway in non small cell lung cancer[J]. Biochem Pharmacol, 2014, 90(3):197-207. DOI:10.1016/j. bcp.2014.05.011.
[18] WU Q, CHEN Y, CUI G, et al. Wortmannin inhibits K562 leukemic cells by regulating PI3k/AKT channel in vitro[J]. J Hua Zhong Univ Sci Technolog Med Sci, 2009, 29(4):451-456. DOI:10.1007/s11596-009-0412-x.
[19] TANG Y, PAN J, HUANG S, et al. Downregulation of miR-133a-3p promotes prostate cancer bone metastasis via activating PI3K/AKT signaling[J]. J Exp Clin Cancer Res, 2018, 37(1):160-166. DOI:10.1186/s13046-018-0813-4.
[20] LI Q, LAI Z, YAN Z, et al. Hedyotis diffusa Willd inhibits proliferation and induces apoptosis of 5-FU resistant colorectal cancer cells by regulating the PI3K/AKT signaling pathway[J]. Mol Med Rep,2018, 17(1):358-365. DOI:10.3892/mmr.2017.7903.
[21] SEO B R, MIN K J, CHO I J, et al. Curcumin significantly enhances dual PI3K/AKT and mTOR inhibitor NVP-BEZ235-induced apoptosis in human renal carcinoma Caki cells through down-regulation of p53-dependent Bcl-2 expression and inhibition of Mcl-1protein stability[J/OL]. PLoS One, 2014, 9(4):e95588[2019-01-12].https://www. ncbi. nlm. nih. gov/pubmed/24743574. DOI:10.1371/journal.pone.0095588.
[22]何平,王平,熊隆信. miR-429通过下调PTEN并激活PI3K/AKT信号通路促进胰腺癌PANC-1细胞对卡培他滨耐药[J].中国肿瘤生物治疗杂志, 2018, 25(12):1251-1258. DOI:10.3872/j.issn.1007-385X.2018.12.007.
[23]赵家义,韩一平. PI3K-AKT-mTOR信号通路抑制剂与肿瘤免疫治疗[J].中国肿瘤生物治疗杂志, 2017, 24(12):1424-1430. DOI:10.3872/j.issn.1007-385X.2017.12.015.
[24] CHEN J, WANG W, WANG H, et al. Combination treatment of ligustrazine piperazine derivate DLJ14 and adriamycin inhibits progression of resistant breast cancer through inhibition of the EGFR/PI3K/AKT survival pathway and induction of apoptosis[J]. Drug Discov Ther, 2014, 8(1):33-41. DOI:10.5582/ddt.8.33.
[2] WANG M, YUANG C, CHANG A. Molecular mechanism of action and potential biomarkers of growth inhibition of synergistic combination of afatinib and dasatinib against gefitinib-resistant non-small cell lung cancer cells[J]. Oncotarget, 2018, 9(23):16533-16546.DOI:10.18632/oncotarget.24814.
[3] BEAUCHAMP E M, WOODS B A, DULAK A M, et al. Acquired resistance to dasatinib in lung cancer cell lines conferred by DDR2gatekeeper mutation and NF1 loss[J]. Mol Cancer Ther, 2014, 13(2):475-482. DOI:10.1158/1535-7163.mct-13-0817.
[4] ZHANG Y, QU X, LI C, et al. miR-103/107 modulates multidrug resistance in human gastric carcinoma by downregulating Cav-1[J].Tumour Biol, 2015, 36(4):2277-2285. DOI:10.1007/s13277-014-2835-7.
[5] WAN L, TIAN Y, ZHANG R, et al. MicroRNA-103 confers the resistance to long-treatment of adriamycin to human leukemia cells by regulation of COP1[J]. J Cell Biochem, 2018, 119(5):3843-3852. DOI:10.1002/jcb.26431.
[6] CHEN H Y, LIN Y M, CHUANG H C, et al. miR-103/107 promote metastasis of colorectal cancer by targeting the metastasis suppressors DAPK and KLF4[J]. Cancer Res, 2017, 77(23):6788-6793.DOI:10.1158/0008-5472.can-17-3079.
[7] YANG D, WANG J J, LI J S, et al. miR-103 functions as a tumor suppressor by directly targeting programmed cell death 10 in NSCLC[J]. Oncol Res, 2017, 26(4):519-528 DOI:10.3727/096504017x15000757094686.
[8] ZHENG J, LIU Y, QIAO Y, et al. miR-103 promotes proliferation and metastasis by targeting KLF4 in gastric cancer[J/OL]. Int J Mol Sci, 2017, 18(5):pii:E910[2019-01-11]. https://www. ncbi. nlm. nih.gov/pubmed/28445396.DOI:10.3390/ijms18050910.
[9] PEREZ-RAMIREZ C, CANADAS-GARRE M, MOLINA M A, et al. PTEN and PI3K/AKT in non-small-cell lung cancer[J]. Pharmacogenomics, 2015, 16(16):1843-1862. DOI:10.2217/pgs.15.122.
[10] ZHANG R, LI G, ZHANG Q, et al. Hirsutine induces m PTP-dependent apoptosis through ROCK1/PTEN/PI3K/GSK3beta pathway in human lung cancer cells[J]. Cell Death Dis, 2018, 9(6):598-605.DOI:10.1038/s41419-018-0641-7.
[11] ZHU J, ZHU WWU W. MicroRNAs change the landscape of cancer resistance[J]. Methods Mol Biol, 2018, 1699(2):83-89. DOI:10.1007/978-1-4939-7435-1_6.
[12] MACDONAGH L, GRAY S G, FINN S P, et al. The emerging role of microRNAs in resistance to lung cancer treatments[J]. Cancer Treat Rev, 2015, 41(2):160-169. DOI:10.1016/j.ctrv.2014.12.009.
[13] WANG C, WANG S, MA F, et al. miRNA328 overexpression confers cisplatin resistance in nonsmall cell lung cancer via targeting of PTEN[J]. Mol Med Rep, 2018, 11(1):625-632 DOI:10.3892/mmr.2018.9478.
[14] SHEN H, ZHU F, LIU J, et al. Alteration in Mir-21/PTEN expression modulates gefitinib resistance in non-small cell lung cancer[J/OL]. PLoS One, 2014, 9(7):e103305[2019-01-11]. http://med.wanfangdata.com.cn/Paper/Detail/PeriodicalPaper_PM25058005.DOI:10.1371/journal.pone.0103305.
[15] WANG N, ZHU C, XU Y, et al. Negative regulation of PTEN by microRNA-221 and its association with drug resistance and cellular senescence in lung cancer cells[J]. Biomed Res Int, 2018, 2018:7908950. https://www. ncbi. nlm. nih. gov/pubmed/29876362. DOI:10.1155/2018/7908950.
[16] ZHANG Q, ZHANG B, SUN L, et al. MicroRNA-130b targets PTEN to induce resistance to cisplatin in lung cancer cells by activating Wnt/beta-catenin pathway[J]. Cell Biochem Funct, 2018, 36(4):194-202. DOI:10.1002/cbf.3331.
[17] FUMAROLA C, BONELLI M A, PETRONINI P G, et al. Targeting PI3K/AKT/mTOR pathway in non small cell lung cancer[J]. Biochem Pharmacol, 2014, 90(3):197-207. DOI:10.1016/j. bcp.2014.05.011.
[18] WU Q, CHEN Y, CUI G, et al. Wortmannin inhibits K562 leukemic cells by regulating PI3k/AKT channel in vitro[J]. J Hua Zhong Univ Sci Technolog Med Sci, 2009, 29(4):451-456. DOI:10.1007/s11596-009-0412-x.
[19] TANG Y, PAN J, HUANG S, et al. Downregulation of miR-133a-3p promotes prostate cancer bone metastasis via activating PI3K/AKT signaling[J]. J Exp Clin Cancer Res, 2018, 37(1):160-166. DOI:10.1186/s13046-018-0813-4.
[20] LI Q, LAI Z, YAN Z, et al. Hedyotis diffusa Willd inhibits proliferation and induces apoptosis of 5-FU resistant colorectal cancer cells by regulating the PI3K/AKT signaling pathway[J]. Mol Med Rep,2018, 17(1):358-365. DOI:10.3892/mmr.2017.7903.
[21] SEO B R, MIN K J, CHO I J, et al. Curcumin significantly enhances dual PI3K/AKT and mTOR inhibitor NVP-BEZ235-induced apoptosis in human renal carcinoma Caki cells through down-regulation of p53-dependent Bcl-2 expression and inhibition of Mcl-1protein stability[J/OL]. PLoS One, 2014, 9(4):e95588[2019-01-12].https://www. ncbi. nlm. nih. gov/pubmed/24743574. DOI:10.1371/journal.pone.0095588.
[22]何平,王平,熊隆信. miR-429通过下调PTEN并激活PI3K/AKT信号通路促进胰腺癌PANC-1细胞对卡培他滨耐药[J].中国肿瘤生物治疗杂志, 2018, 25(12):1251-1258. DOI:10.3872/j.issn.1007-385X.2018.12.007.
[23]赵家义,韩一平. PI3K-AKT-mTOR信号通路抑制剂与肿瘤免疫治疗[J].中国肿瘤生物治疗杂志, 2017, 24(12):1424-1430. DOI:10.3872/j.issn.1007-385X.2017.12.015.
[24] CHEN J, WANG W, WANG H, et al. Combination treatment of ligustrazine piperazine derivate DLJ14 and adriamycin inhibits progression of resistant breast cancer through inhibition of the EGFR/PI3K/AKT survival pathway and induction of apoptosis[J]. Drug Discov Ther, 2014, 8(1):33-41. DOI:10.5582/ddt.8.33.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |