lncRNA FOXD2-AS1通过调控miR-185-5p/CCND2分子轴诱导胃癌细胞MGC-803对阿帕替尼的耐药性
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摘要
目的:探讨lncRNA FOXD2-AS1(FOXD2-AS1)通过调控miR-185-5p/CCND2分子轴参与胃癌细胞对阿帕替尼耐药性的分子机制。方法:收集无锡市第五医院2016年4月至2017年12月间收治的资料完整的25例胃癌患者癌组织和相应癌旁组织标本,采用qRT-PCR检测FOXD2-AS1、miR-185-5p和CCND2在胃癌组织或细胞系中的表达水平;采用CCK-8、Transwell和Annexin V-FITC/PI双染流式术检测胃癌细胞对阿帕替尼药物的敏感性;采用双荧光素酶报告基因验证FOXD2-AS1、miR-185-5p和CCND2的靶向关系,并通过Western blotting和qRT-PCR检测其调控关系。结果:FOXD2-AS1在胃癌组织和阿帕替尼耐药细胞株中高表达;同时,过表达FOXD2-AS1可促进胃癌MGC-803/AP细胞对阿帕替尼的耐药性。双荧光素酶报告基因证实FOXD2-AS1靶向作用miR-185-5p并下调其表达水平。miR-185-5p通过抑制胃癌MGC-803/AP细胞增殖、侵袭和促进凋亡进而下调FOXD2-AS1对胃癌细胞阿帕替尼耐药性的促进作用。miR-185-5p可靶向负调控CCND2的表达,FOXD2-AS1通过下调miR-185-5p对CCND2的抑制作用进而促进胃癌MGC-803/AP细胞增殖、侵袭和抑制凋亡,从而上调胃癌细胞对阿帕替尼的耐药性。结论:FOXD2-AS1通过调控miR-185-5p/CCND2分子轴诱导胃癌细胞对阿帕替尼的耐药性。
Objective: To investigate the molecular mechanism of lncRNA FOXD2-AS1 participating in apatinib resistance in gastric cancer cells by regulating miR-185-5 p/CCND2 axis. Methods: The gastri cancer tissues and corresponding paracancerous tissues of 25 patients with gastric cancer were collected from April 2016 to December 2017 in the Fifth People's Hospital of Wuxi City. The expressions of FOXD2-AS1, miR-185-5 p, and cyclin D2(CCND2) in gastric cancer tissues or cell lines were examined by quantitative realtime polymerase chain reaction(qRT-PCR). CCK-8 assay, Transwell assay and Annexin V-FITC/PI double staining flow cytometry assay were applied to assess the sensitivity of gastric cancer cells to apatinib. The interaction between FOXD2-AS1, miR-185-5 p and CCND2 was explored by dual luciferase reporter gene assay, which was then confirmed by qRT-PCR, and Western blotting. Results:FOXD2-AS1 was highly expressed in gastric cancer tissues and apatinib-resistant gastric cancer cells. Over-expression of FOXD2-AS1 promoted apatinib-resistance of MGC-803/AP cells. Dual luciferase reporter gene assay confirmed that FOXD2-AS1 directly interacted with miR-185-5 p and suppressed its expression. miR-185-5 p significantly abolished the promotion effect of FOXD2-AS1 on apatinibresistance via inhibiting cell proliferation, invasion and promoting apoptosis of gastric MGC-803/AP cells. miR-185-5 p could negatively regulate CCND2 expression; and FOXD2-AS1 promoted the cell proliferation, invasion and inhibited apoptosis of MGC-803/AP cells via down-regulating the inhibition effect of miR-185-5 p on CCND2, thus further enhanced the apatinib-resistance of gastric cancer cells. Conclusion: FOXD2-AS1 induced apatinib-resistance of gastric cancer cells by regulating miR-185-5 p/CCND2 axis.
Objective: To investigate the molecular mechanism of lncRNA FOXD2-AS1 participating in apatinib resistance in gastric cancer cells by regulating miR-185-5 p/CCND2 axis. Methods: The gastri cancer tissues and corresponding paracancerous tissues of 25 patients with gastric cancer were collected from April 2016 to December 2017 in the Fifth People's Hospital of Wuxi City. The expressions of FOXD2-AS1, miR-185-5 p, and cyclin D2(CCND2) in gastric cancer tissues or cell lines were examined by quantitative realtime polymerase chain reaction(qRT-PCR). CCK-8 assay, Transwell assay and Annexin V-FITC/PI double staining flow cytometry assay were applied to assess the sensitivity of gastric cancer cells to apatinib. The interaction between FOXD2-AS1, miR-185-5 p and CCND2 was explored by dual luciferase reporter gene assay, which was then confirmed by qRT-PCR, and Western blotting. Results:FOXD2-AS1 was highly expressed in gastric cancer tissues and apatinib-resistant gastric cancer cells. Over-expression of FOXD2-AS1 promoted apatinib-resistance of MGC-803/AP cells. Dual luciferase reporter gene assay confirmed that FOXD2-AS1 directly interacted with miR-185-5 p and suppressed its expression. miR-185-5 p significantly abolished the promotion effect of FOXD2-AS1 on apatinibresistance via inhibiting cell proliferation, invasion and promoting apoptosis of gastric MGC-803/AP cells. miR-185-5 p could negatively regulate CCND2 expression; and FOXD2-AS1 promoted the cell proliferation, invasion and inhibited apoptosis of MGC-803/AP cells via down-regulating the inhibition effect of miR-185-5 p on CCND2, thus further enhanced the apatinib-resistance of gastric cancer cells. Conclusion: FOXD2-AS1 induced apatinib-resistance of gastric cancer cells by regulating miR-185-5 p/CCND2 axis.
引文
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[2]JIANG L,YANG W,BIAN W,et al.microRNA-623 targets cyclin D1 to inhibit cell proliferation and enhance the chemosensitivity of cells to 5-fluorouracil in gastric cancer[J].Oncol Res,2018.DOI:10.3727/096504018x15193469240508.
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[4]WANG Z,JI F.Downregulation of microRNA-17-5p inhibits drug resistance of gastric cancer cells partially through targeting p21[J].Oncol Lett,2018,15(4):4585-4591.DOI:10.3892/ol.2018.7822.
[5]WANG X,ZHANG H,BAI M,et al.Exosomes serve as nanoparticles to deliver anti-miR-214 to reverse chemoresistance to cisplatin in gastric cancer[J].Mol Ther,2018,26(3):774-783.DOI:10.1016/j.ymthe.2018.01.001.
[6]WANG L,MA L,XU F,et al.Role of long non-coding RNA in drug resistance in non-small cell lung cancer[J].Thorac Cancer,2018,9(7):761-768.DOI:10.1111/1759-7714.12652.
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[12]CHEN Q N,WEI C C,WANG Z X,et al.Long non-coding RNAs in anti-cancer drug resistance[J].Oncotarget,2017,8(1):1925-1936.DOI:10.18632/oncotarget.12461.
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[27]CHEN Y,ZHANG Q,WANG Q,et al.Genetic association analysis of the RTK/ERK pathway with aggressive prostate cancer highlights the potential role of CCND2 in disease progression[J].Sci Rep,2017,7(1):4538.DOI:10.1038/s41598-017-04731-4.
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[29]WANG Y,XUE J,KUANG H,et al.microRNA-1297 inhibits the growth and metastasis of colorectal cancer by suppressing cyclin D2 expression[J].DNA Cell Biol,2017,36(11):991-999.DOI:10.1089/dna.2017.3829.
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