iR-17在非小细胞肺癌放射抗拒细胞中的表达及作用研究

doi:10.11904/j.issn.1002-3070.2018.05.002

实用肿瘤学杂志 ›› 2018, Vol. 32 ›› Issue (5): 391-397.doi: 10.11904/j.issn.1002-3070.2018.05.002

iR-17在非小细胞肺癌放射抗拒细胞中的表达及作用研究

李万祯, 殷俊, 李萍, 林晓辉, 张国前, 张书旭

广州医科大学附属肿瘤医院放疗中心(广州 510095)

收稿日期:2018-04-26 出版日期:2018-10-28 发布日期:2018-10-23
通讯作者: 张书旭,E-mail:gthzsx@163.comm
作者简介:李万祯,男,(1990-),硕士研究生,从事肿瘤放射生物及放射物理的研究。
基金资助:
广东省科技计划项目(编号:2016A020215173)

The expression and role of miR-17 in radioresistant cells of non-small cell lung cancer

LI Wanzhen, YIN Jun, LI Ping, LIN Xiaohui, ZHANG Guoqian, ZHANG Shuxu

Radiotherapy Center,Affiliated Cancer Hospital & Institute of Guangzhou Medical University,Guangzhou 510095,China

Received:2018-04-26 Online:2018-10-28 Published:2018-10-23

摘要/Abstract

摘要： 目的建立肺癌放射抗拒细胞系H1975R,并探究miR-17在其放射抗拒中的作用。方法通过梯度剂量连续照射,体外建立肺癌放射抗拒细胞系H1975R。采用克隆形成实验对H1975R和H1975细胞的放射敏感性进行验证。利用RT-PCR检测亲本细胞和放射抗拒细胞的miR-17的表达水平,并通过上调或下调miR-17表达,比较其放射敏感性变化,应用CCK-8法检测细胞增殖,流式细胞术检测细胞凋亡和周期分布。结果 H1975R的放射生物学参数D₀、D_q和SF₂均比亲本H1975大(P＜0.05),显示更强的放射抗拒性。从RT-PCR结果可知,miR-17在H1975R的表达量是H1975的2.67倍(P＜0.01)。上调H1975细胞的miR-17表达后,其放射敏感性降低,而细胞增殖和抗凋亡能力均增强,G₂/M期细胞比例增多(P＜0.05)。下调H1975R细胞的miR-17表达后,其放射敏感性升高,而细胞增殖和抗凋亡能力均降低,G₂/M期细胞比例减少(P＜0.05)。结论 miR-17参与非小细胞肺癌放射抗拒的调控,其可能成为治疗放射抗拒的新靶点。

关键词: 非小细胞肺癌, 放射抗拒, 增殖, 凋亡, miR-17

Abstract: Objective The aims of this study were to establish a radioresistant human lung cancer cell line H1975R,and to explore the role of miR-17 in the formation of radioresistance.Methods The lung cancer radioresistant cell line H1975R was established in vitro by continuous irradiation with gradient dose.The radiosensitivity of H1975R and H1975 cells was verified by colony formation assay.We determined the expression levels of miR-17 in parental cells and radioresistant cells by quantitative RT-PCR and The expression of miR-17 was up-regulated or down-regulated,and the changes in radiosensitivity were compared.Cell proliferation was detected by CCK-8 assay.Flow cytometry was performed to determine apoptosis and cycle distribution.Results The radiobiological parameters(D₀,N,D_q and SF₂)of H1975R cells were greater than those in the H1975 cells(P＜0.05),indicating a stronger radioresistance.From the Results of RT-PCR,the expression level of miR-17 in H1975R cells was 2.67 times than that in H1975 cells(P＜0.01).After up-regulating expression of miR-17 in H1975 cells,its radiosensitivity was reduced,but the proliferation,anti-apoptosis and the proportion of cells in the G₂/M phase increased(P＜0.05).Similarly,after down-regulating expression of miR-17 in H1975R cells,the radiosensitivity increased,while the cell proliferation and anti-apoptosis ability decreased,and the proportion of cells in the G₂/M phase decreased(P＜0.05).Conclusion miR-17 is involved in the regulation of radioresistance in non-small cell lung cancer,which may become a new target for the treatment of radioresistance.

Key words: Non-small cell lung cancer, Radioresistance, Proliferation, Apoptosis, miR-17

中图分类号:

R734.2

李万祯, 殷俊, 李萍, 林晓辉, 张国前, 张书旭. iR-17在非小细胞肺癌放射抗拒细胞中的表达及作用研究[J]. 实用肿瘤学杂志, 2018, 32(5): 391-397.

LI Wanzhen, YIN Jun, LI Ping, LIN Xiaohui, ZHANG Guoqian, ZHANG Shuxu. The expression and role of miR-17 in radioresistant cells of non-small cell lung cancer[J]. PRACTICAL ONCOLOGY JOURNAL, 2018, 32(5): 391-397.

参考文献

1 Chen W,Zheng R,Zhang S,et al.Cancer incidence and mortality in China in 2013:an analysis based on urbanization level[J].Chin J Cancer Res,2017,29(1):1-10.
2 Siegel RL,Miller KD,Jemal A.Cancer statistics,2016[J].CA Cancer J Clin,2016,66(1):7-30.
3 Xie S,Li G,Han C,et al.RKIP reduction enhances radioresistance by activating the Shh signaling pathway in non-small-cell lung cancer[J].Onco Targets Ther,2017,10:5605-5619.
4 Le Pechoux C.Role of postoperative radiotherapy in resected non-small cell lung cancer:a reassessment based on new data[J].Oncologist,2011,16(5):672-681.
5 Luo M,Ding L,Li Q,et al.miR-668 enhances the radioresistance of human breast cancer cell by targeting IκBα[J].Breast Cancer,2017,24(5):673-682.
6 Kim J,Chang JW,Yun HS,et al.Chloride intracellular channel 1 identified using proteomic analysis plays an important role in the radiosensitivity of HEp-2 cells via reactive oxygen species production[J].Proteomics,2010,10(14):2589-2604.
7 Li G,Liu Y,Su Z,et al.MicroRNA-324-3p regulates nasopharyngeal carcinoma radioresistance by directly targeting WNT2B[J].Eur J Cancer,2013,49(11):2596-2607.
8 Zhao RS,Wang W,Li JP,et al.Gelsolin promotes radioresistance in non-small cell lung cancer cells through activation of phosphoinositide 3-Kinase/Akt signaling[J].Technol Cancer Res Treat,2017,16(4):512-518.
9 Schou JV,Johansen JS,Nielsen D,et al.Circulating microRNAs as prognostic and predictive biomarkers in patients with colorectal cancer[J].Noncoding RNA,2016,2(2):5.
10 Zhou P,Ma L,Zhou J,et al.miR-17-92 plays an oncogenic role and conveys chemo-resistance to cisplatin in human prostate cancer cells[J].Int J Oncol,2016,48(4):1737-1748.
11 Jiang P,Rao EY,Meng N,et al.MicroRNA-17-92 significantly enhances radioresistance in human mantle cell lymphoma cells[J].Radiat Oncol,2010,5:100.
12 Mahmood J,Zaveri SR,Murti SC,et al.Caveolin-1:a novel prognostic biomarker of radioresistance in cancer[J].Int J Radiat Biol,2016,92(12):747-753.
13 Wei K,Kodym R,Jin C.Radioresistant cell strain of human fibrosarcoma cells obtained after long-term exposure to X-rays[J].Radiat Environ Biophys,1998,37(2):133-137.
14 Pearce AG,Segura TM,Rintala AC,et al.The generation and characterization of a radiation-resistant model system to study radioresistance in human breast cancer cells[J].Radiat Res,2001,156(6):739-750.
15 殷蔚伯,余子豪,徐国镇,等.肿瘤放射治疗学[M].北京:中国协和医科大学出版社,2008:231-237.
16 Reddy KB.MicroRNA(miRNA)in cancer[J].Cancer Cell International,2015,15(1):1-6.
17 Wu SY,Lin KC,Chiou JF,et al.MicroRNA-17-5p post-transcriptionally regulates p21 expression in irradiated betel quid chewing-related oral squamous cell carcinoma cells[J].Strahlenther Onkol,2013,189(8):675-683.
18 Hayashita Y,Osada H,Tatematsu Y,et al.A polycistronic microRNA cluster,miR-17-92,is overexpressed in human lung cancers and enhances cell proliferation[J].Cancer Res,2005,65(21):9628-9632.
19 刘国慧,谷安鑫,殷洪涛,等.原苏木素A对胃癌SGC-7901细胞系放疗增敏作用的研究[J].实用肿瘤学杂志,2017,31(6):500-505.
20 Yao Y,Cui Y,Qiu X,et al.Attenuated LKB1-SIK1 signaling promotes epithelial-mesenchymal transition and radioresistance of non-small cell lung cancer cells[J].Chin J Cancer,2016,35:50.

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iR-17在非小细胞肺癌放射抗拒细胞中的表达及作用研究

The expression and role of miR-17 in radioresistant cells of non-small cell lung cancer

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