VEGF基因沉默对人肺癌A549细胞放射增敏作用的体内研究

doi:10.3969/j.issn.1674-5817.2019.04.005

实验动物与比较医学 ›› 2019, Vol. 39 ›› Issue (4): 280-286.DOI: 10.3969/j.issn.1674-5817.2019.04.005

VEGF基因沉默对人肺癌A549细胞放射增敏作用的体内研究

马军¹, 罗居东², 景文江¹, 张淑莲¹, 王娟毅¹, 范志刚¹

1.汉中三二〇一医院肿瘤科, 汉中 723000;
2.南京医科大学附属常州第二人民医院肿瘤中心, 常州 213164

收稿日期:2019-01-10 出版日期:2019-08-25 发布日期:2021-01-31
作者简介:马军(1971-),男,主治医师,研究方向:肿瘤。E-mail:ximajun@163.com
基金资助:
江苏省科技厅重点研发项目(BE2018643)

Investigation on Radiosensitivity Impact of Lung Cancer A549 Cells by Vascular Endothelial Growth Factor Silencing in vivo

MA Jun¹, LUO Jun-dong², JING Wen-jiang¹, ZHANG Shu-lian¹, WANG Juan-yi¹, FAN Zhi-gang¹

1. Department of Oncology, Hanzhong 3201 Hospital, Hanzhong 723000, China;
2. Tumor Center,Changzhou Second People's Hospital Affiliated to Nanjing Medical University, Changzhou 213164, China

Received:2019-01-10 Online:2019-08-25 Published:2021-01-31

摘要/Abstract

摘要： 目的探讨RNA干扰血管内皮生长因子(VEGF)基因表达载体siVEGF对人肺腺癌A549细胞荷瘤鼠的放射敏感性的影响。方法用体外培养人肺腺癌A549细胞株建立荷瘤裸小鼠模型,荷瘤鼠瘤内注射siVEGF/RNAi-MateTM脂质体混合物,再经放射治疗24 d后,处死裸小鼠,剥离瘤体称重,利用实时定量PCR (q-PCR)、免疫组织化学和Western blotting检测VEGF的基因和蛋白表达水平,评价siVEGF对放射敏感性的影响。结果 q-PCR、Western blotting和免疫组织化学检测结果表明siVEGF可显著降低VEGF的蛋白水平(P<0.05)。与对照组相比siVEGF联合放疗组的肿瘤生长和坏死程度存在显著性差异(P<0.05)。结论 siVEGF合并放射治疗能显著抑制A549细胞移植瘤的生长,增加其对放射治疗的敏感性。

关键词: 人肺腺癌, A549细胞, 沉默RNA, 血管内皮生长因子(VEGF), 放射敏感性

Abstract: Objective To research the radiosensitivity of A549 cells in tumor-bearing mouse model by knock down vascular endothelial growth factor (VEGF) gene. Methods The tumor-bearing mouse model was established by using A549 cells which was cultured in vitro. The siVEGF/RNAi-MateTM mixture was injected into the tumor of model mice and with radiotherapy. The nude mice were euthanasia on day 24 after radiotherapy. The expressions of VEGF were detected by immuno-histochemical, Q-PCR and Western blotting. Results Both the protein and mRNA levels of VEGF in VEGF knockdown and radiotherapy group were significantly decreased compared with the control group (P<0.05). The volume of tumor was decreased and the necrotic area of tumor were increased distinctly in siVEGF transfected and radiotherapy group compared with control group (P<0.05). Conclusion Knock down of VEGF and radiotherapy can inhibit tumor growth, and increase the radiosensitivity of lung cancer radiotherapy.

Key words: Lung cancer, A549, siRNA, Vascular endothelial growth factor (VEGF), Radiosensitivity

中图分类号:

Q95-33

马军, 罗居东, 景文江, 张淑莲, 王娟毅, 范志刚. VEGF基因沉默对人肺癌A549细胞放射增敏作用的体内研究[J]. 实验动物与比较医学, 2019, 39(4): 280-286.

MA Jun, LUO Jun-dong, JING Wen-jiang, ZHANG Shu-lian, WANG Juan-yi, FAN Zhi-gang. Investigation on Radiosensitivity Impact of Lung Cancer A549 Cells by Vascular Endothelial Growth Factor Silencing in vivo[J]. Laboratory Animal and Comparative Medicine, 2019, 39(4): 280-286.

参考文献

[1] Allemani C, Weir HK, Carreira H, et al.Global surveillance of cancer survival 1995-2009: Analysis of individual data for 25 676 887 patients from 279 population-based registries in 67 countries (concord-2)[J]. The Lancet, 2015, 385(9972):977-1010.
[2] 陈万青, 李贺, 孙可欣, 等. 2014年中国恶性肿瘤发病和死亡分析[J]. 中华肿瘤杂志, 2018, 40(1):5-13.
[3] Siegel RL, Miller KD, Jemal A.Cancer statistics, 2019[J]. CA Cancer J Clin, 2019, 69(1):7-34.
[4] 郑荣寿, 孙可欣, 张思维, 等. 2015年中国恶性肿瘤流行情况分析[J]. 中华肿瘤杂志, 2019, 41(1):19028.
[5] Ferlay J, Colombet M, Soerjomataram I, et al.Cancer incidence and mortality patterns in europe: Estimates for 40 countries and 25 major cancers in 2018[J]. Eur J Cancer, 2018, 103(8):358-387.
[6] Islami F, Chen W, Yu XQ, et al.Cancer deaths and cases attributable to lifestyle factors and infections in China, 2013[J]. Ann Oncol, 2017, 28(10):2567-2574.
[7] 陈万青, 郑荣寿, 张思维, 等. 2013年中国老年人群恶性肿瘤发病和死亡分析[J]. 中华肿瘤杂志, 2017, 39(2):60-67.
[8] 陈万青, 左婷婷, 郑荣寿, 等. 2013年中国肺癌发病与死亡分析[J]. 中华肿瘤杂志, 2017, 39(10):795-800.
[9] O'Rourke N, Roqué I Figuls M, et al. Concurrent chemoradiotherapy in non-small cell lung cancer[J]. Cochrane Database Syst Rev, 2010, 16(6):CD002140.
[10] Kufe D, Weichselbaum R.Radiation therapy: activation of gene transcription and the development of genetic radiotherapy[J]. Cancer Biol Ther, 2003, 2:326-329.
[11] Li X, Guo X, Cheng Y, et al.pH-responsive cross-linked low molecular weight polyethylenimine as an efficient gene vector for delivery of plasmid DNA encoding anti-VEGF-shRNA for tumor treatment[J]. Front Oncol, 2018, 8(1):354.
[12] Lyons Y, Pradeep S, Wu S, et al.Macrophage depletion through colony stimulating factor 1 receptor pathway blockade overcomes adaptive resistance to anti-VEGF therapy[J]. Oncotarget, 2017, 8(57):96496-96505.
[13] Ryoo NK, Lee J, Lee H, et al.Therapeutic effects of a novel siRNA-based anti-VEGF (siVEGF) nanoball for the treatment of choroidal neovascularization[J]. Nanoscale, 2017, 9(40):15461-15469.
[14] Zhang L, Yang N, Mohamed-Hadley A, et al.Vector-based RNAi, a novel tool for isoform-specific knock-down of VEGF and anti-angiogenesis gene therapy of cancer[J]. Biochem Biophys Res Commun, 2003, 303(4):1169-1178.
[15] Tudoran O, Berindan neagoe I, Balacescu O, et al. Evaluation of tumor angiogenesis through VEGF modulation in ovarian cancerin vivousing RNA interference[J]. Rom Biotech Lett,2009, 14(4):4560-4566.
[16] Zhu H, Yun F, Shi X, et al.VEGF-C inhibition reverses resistance of bladder cancer cells to cisplatin via upregulating maspin[J]. Mol Med Rep, 2015, 12(2):3163-3169.
[17] Matsumoto G, Kushibiki T, Kinoshita Y, et al.Cationized gelatin delivery of a plasmid DNA expressing small interference RNA for VEGF inhibits murine squamous cell carcinoma[J]. Cancer Sci, 2006, 97(4):313-321.
[18] Ohta Y, Endo Y, Tanaka M, et al.Significance of vascular endothelial growth factor messenger RNA expression in primary lung cancer[J]. Clin Cancer Res, 1996, 2(8):1411-1416.
[19] Senan S, Smit EF.Design of clinical trials of radiation combined with antiangiogenic therapy[J]. Oncologist, 2007, 12(4):465-477.
[20] Zhu YM, Azahri NS, Yu DC, et al.Effects of COX-2 inhibition on expression of vascular endothelial growth factor and interleukin-8 in lung cancer cells[J]. BMC Cancer, 2008, 8:218.
[21] Rich JN.Cancer stem cells in radiation resistance[J]. Cancer Res, 2007, 67(19):8980-89804.
[22] Nur SM, Hasan MA, Amin MA, et al.Design of potential RNAi (miRNA and siRNA) molecules for middle east respiratory syndrome coronavirus (MERS-CoV) gene silencing by computational method[J]. Interdiscip Sci, 2015, 7(3):257-265.
[23] Fire A, Xu S, Montgomery MK, et al.Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans[J]. Nature, 1998, 391(6669):806-811.
[24] Jie Z, Mo C, Cai-cun Z, et al. Influences of over-expression of VEGF-C and its inhibition on biological characteristics of NCI-H446 human small-cell lung cancer cells[J]. Tumor, 2012, 32(2):11-18.
[25] Barr MP, Gray SG, Gately K, et al.Vascular endothelial growth factor is an autocrine growth factor, signaling through neuropilin-1 in non-small cell lung cancer[J]. Mol Cancer, 2015, 14:45.
[26] Zhou HB, Yin YF, Hu Y, et al.Suppression of vascular endothelial growth factor via siRNA interference modulates the biological behavior of human nasopharyngeal carcinoma cells[J]. Jpn J Radiol, 2011, 29(9):615-622.
[27] Lecaros RL, Huang L, Lee TC, et al.Nanoparticle delivered VEGF-A siRNA enhances photodynamic therapy for head and neck cancer treatment[J]. Mol Ther, 2015, 169:65-74.

VEGF基因沉默对人肺癌A549细胞放射增敏作用的体内研究

Investigation on Radiosensitivity Impact of Lung Cancer A549 Cells by Vascular Endothelial Growth Factor Silencing in vivo

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价

[1]	陈林中日, 殷冬来, 赵泽婷, 韦芊含, 陈昕, 张雨梅. 靶向大鼠Vegf/Vegfr基因siRNA的基因沉默效应研究[J]. 实验动物与比较医学, 2019, 39(1): 26-33.
[2]	唐彬, 黎福荣, 付新录, 江国建, 赵勇, 张薇. 米非司酮对Wistar大鼠窗口期子宫内膜整合素和血管内皮生长因子基因表达的影响[J]. 实验动物与比较医学, 2017, 37(5): 399-404.