氟尿嘧啶调控VHL/HIF信号通路抑制肾癌细胞增殖、侵袭与转移的研究

doi:10.12092/j.issn.1009-2501.2019.08.005

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (8): 872-879.doi: 10.12092/j.issn.1009-2501.2019.08.005

氟尿嘧啶调控VHL/HIF信号通路抑制肾癌细胞增殖、侵袭与转移的研究

周青英¹，徐乃奋¹，郑定钦¹，潘其壮¹，张志刚¹，汪朔²

¹平阳人民医院泌尿外科，温州 325400，浙江；²浙江大学医学院附属第一医院泌尿外科，杭州 310003，浙江

收稿日期:2018-11-28 修回日期:2019-06-20 出版日期:2019-08-26 发布日期:2019-08-30
通讯作者: 汪朔，男，硕士，主任医师，教授，研究方向：肾癌、前列腺癌的基因靶向、免疫、化学治疗。
作者简介:周青英，男，本科，主任医师，研究方向：肾癌、前列腺癌的免疫、化学治疗。 E-mail：20412077@qq.com
基金资助:
浙江省卫生厅项目（2019KY676）

Fluorouracil regulates VHL/HIF signaling pathway and inhibits proliferation, invasion and metastasis of renal carcinoma cells

ZHOU Qingying¹, XU Naifen¹, ZHENG Dingqin¹, PAN Qizhuang¹, ZHANG Zhigang¹, WANG Shuo²

¹Department of Urology, Pingyang People's Hospital, Wenzhou 325400, Zhejiang, China; ²Department of Urology, the First Affiliated Hospital of Medical College, Zhejiang University, Hangzhou 310003, Zhejiang, China

Received:2018-11-28 Revised:2019-06-20 Online:2019-08-26 Published:2019-08-30

摘要/Abstract

摘要：

目的:探讨氟尿嘧啶通过调控VHL/HIF信号通路影响肾癌细胞增殖、侵袭与转移。方法:体外培养人肾癌786-0细胞，分别用终浓度为6、60、600 μg/mL的氟尿嘧啶处理肾癌细胞，噻唑蓝（MTT）检测氟尿嘧啶对肾癌细胞增殖的影响，划痕实验检测细胞转移能力的影响，Transwell实验检测细胞体外侵袭能力的影响。Western blot检测细胞中VHL/HIF信号通路相关蛋白HIF-1α、VEGF及活化的含半胱氨酸的天冬氨酸蛋白水解酶3（Cleaved Caspase-3）、基质金属蛋白酶-9（MMP-9）的表达。通过细胞转染法转染pcDNA-HIF-1α后Western blot检测转染效率，同样的方法检测对肾癌细胞增殖、侵袭和转移的影响。结果:氟尿嘧啶处理后的肾癌细胞增殖明显受到抑制，侵袭和转移能力明显降低，且呈现出时间和浓度依赖性（P<0.05），相关蛋白HIF-1α、VEGF、MMP-9表达下调，Cleaved Caspase-3表达上调，转染pcDNA-HIF-1α后，HIF-1α的表达上调，肾癌细胞增殖活性降低,侵袭和转移能力减弱，与对照组相比，差异具有统计学意义（P<0.05）。结论:氟尿嘧啶能抑制肾癌细胞增殖、侵袭与转移，其作用机制与通过阻断VHL/HIF信号通路，调节Cleaved Caspase-3和MMP-9的表达有关。

关键词: VHL/HIF信号通路, 肾癌细胞, 增殖, 侵袭, 转移

Abstract:

AIM: To investigate the effect of fluorouracil on the proliferation, invasion and metastasis of renal cell carcinoma by regulating VHL/HIF signaling pathway. METHODS: Human kidney cancer 786-0 cells were cultured in vitro. Renal cancer cells were treated with fluorouracil with final concentrations of 6, 60, and 600 μg/mL, respectively. Effects of fluorouracil against kidney cancer cells proliferation was detected by MTT assay. Scratch test was applied to detect the effect of fluorouracil on the metastasis capacity of renal cell carcinoma, and Transwell assay to detect the effect of uracil on the invasion ability of renal cell carcinoma in vitro. Western blot analyzed the VHL/HIF signaling pathway-associated protein HIF-1α, VEGF, activated cysteine-containing aspartate proteinase 3 (Cleaved Caspase-3), matrix metalloproteinases in renal cell carcinoma cells treated with fluorouracil and the effect of 9 (MMP-9) expression. After transfection of pcDNA-HIF-1α by the cell transfection method, the transfection efficiency was detected by Western blot, and the same method was used to examine the effect on the proliferation, invasion and metastasis of renal cell carcinoma.RESULTS:The proliferation of renal cell carcinoma cells treated with fluorouracil was significantly inhibited, and the invasion and metastasis ability was significantly decreased, and it was time-and concentration-dependent (P<0.05). The expression of related proteins HIF-1α, VEGF, and MMP-9 was down-regulated, and Cleaved Caspase-3 the expression was up-regulated. After transfected with pcDNA-HIF-1α, the expression of HIF-1α is upregulated. The proliferation activity and invasion and metastasis ability of renal cell carcinoma were weakened. Compared with the control group, the difference was statistically significant (P<0.05). CONCLUSION:Fluorouracil can inhibit the proliferation, invasion and metastasis of renal cancer cells, and its mechanism is related to blocking the VHL/HIF signaling pathway and regulating the expression of Cleaved Caspase-3 and MMP-9.

Key words: VHL/HIF signaling pathway, renal cancer cells, proliferation, invasion, metastasis

中图分类号:

R965.2

周青英，徐乃奋，郑定钦，潘其壮，张志刚，汪朔. 氟尿嘧啶调控VHL/HIF信号通路抑制肾癌细胞增殖、侵袭与转移的研究[J]. 中国临床药理学与治疗学, 2019, 24(8): 872-879.

ZHOU Qingying, XU Naifen, ZHENG Dingqin, PAN Qizhuang, ZHANG Zhigang, WANG Shuo. Fluorouracil regulates VHL/HIF signaling pathway and inhibits proliferation, invasion and metastasis of renal carcinoma cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(8): 872-879.

参考文献

［1］Fairlamb DJ.Spontaneous regression of metastases of renal cancer: A report of two cases including the first recorded regression following irradiation of a dominant metastasis and review of the world literature［J］. Cancer, 2015, 47(8):2102-2106.
［2］Greef B, Eisen T. Medical treatment of renal cancer: new horizons［J］. Brit J Cancer, 2016, 115(5):505-516.
［3］杨康, 李魏芳, 喻俊峰. 靶向免疫检查点治疗肾细胞癌的进展［J］.实用医学杂志, 2016, 32(7):1068-1069.
［4］Asadi E, Abdouss M, Leblanc RM, et al. In vitro/in vivo study of novel anti-cancer, biodegradable cross-linked tannic acid for fabrication of 5-fluorouracil-targeting drug delivery nano-device based on a molecular imprinted polymer［J］. Rsc Advances, 2016, 6(43):37308-37318.
［5］Xie C, Fu L, Han Y, et al.Decreased ARID1A expression facilitates cell proliferation and inhibits 5-fluorouracil-induced apoptosis in colorectal carcinoma［J］.Tumor Biol, 2014, 35(8):7921-7927.
［6］Lou S, Ren L, Xiao J, et al. Expression profiling based graph-clustering approach to determine renal carcinoma related pathway in response to kidney cancer［J］. Eur Rev Med Pharmacol Sci, 2012, 16(6):775-780.
［7］Boguslawska J, Kedzierska H, Poplawski P, et al. Expression of genes involved in cellular adhesion and extracellular matrix remodeling correlates with poor survival of patients with renal cancer［J］. J Urology, 2016, 195(6):1892-1902.
［8］Schdel J, Grampp S, Maher ER, et al. Hypoxia, Hypoxia-inducible transcription factors, and renal cancer［J］. Eur Urol, 2016, 69(4):646-657.
［9］聂明秀, 张洪秀, 李伟. VHL基因和HIF缺氧反应对肾癌的研究进展［J］. 昆明理工大学学报(自然科学版), 2015, 6(3):101-105.
［10］郭放, 郑振东, 谢晓冬. 晚期肾癌分子靶向治疗新进展［J］. 中国肿瘤临床, 2016, 43(22):977-980.
［11］聂磊, 柏帅, 刘卫,等. 氟尿嘧啶和奥沙利铂对消化道肿瘤的协同抑制作用［J］. 实用医药杂志, 2014, 31(1):40-41.
［12］Dobritzsch D, Schneider G, Schnackerz KD, et al. Crystal structure of dihydropyrimidine dehydrogenase, a major determinant of the pharmacokinetics of the anti-cancer drug 5-fluorouracil［J］. Embo J, 2014, 20(4):650-660.
［13］Peter B, Lili D, Szilvia J, et al. The PCNA-associated protein PARI negatively regulates homologous recombination via the inhibition of DNA repair synthesis［J］. Nucleic Acids Res, 2016, 44(7):3176-3189.
［14］Watanabe M, Nagai Y, Kinoshita K, et al. Induction Chemotherapy with Docetaxel/Cisplatin/5-Fluorouracil for Patients with Node-Positive Esophageal Cancer［J］. Digestion, 2011, 83(3):146-152.
［15］Samatiwat P, Prawan A, Senggunprai L, et al. Repression of Nrf2 enhances antitumor effect of 5-fluorouracil and gemcitabine on cholangiocarcinoma cells［J］. N-S Arch Pharmacol, 2015, 388(6):601-612.
［16］Ardalan B, Glazer RI, Kensler TW, et al. Synergistic effect of 5-fluorouracil and, ja:math, on cell growth and ribonucleic acid synthesis in a human mammary carcinoma［J］. Biochem Pharmacol, 2015, 30(15):2045-2049.
［17］Damjanovic SS, Ilic BB, Cokic BB, et al. Tuberous sclerosis complex protein 1 expression is affected by VHL Gene alterations and HIF-1α production in sporadic clear-cell renal cell carcinoma［J］. Exp Mol Pathol, 2016, 101(3):323-331.
［18］Shen C, Kaelin WG Jr. The VHL/HIF axis in clear cell renal carcinoma［J］. Semin Cancer Biol, 2013, 23(1):18-25.
［19］Shareef MM, Udayakumar TS, Sinha VK, et al. Interaction of HIF-1α and Notch3 Is required for the expression of carbonic anhydrase 9 in breast carcinoma cells［J］. Genes & Cancer, 2013, 4(11/12):513-523.
［20］Schnenberger D, Harlander S, Rajski M, et al. Formation of renal cysts and tumors in Vhl/Trp53-deficient mice requires HIF-1α and HIF-2α［J］. Cancer Res, 2016, 76(7):2025-2036.
［21］贾云波, 杜惠兰, 高星,等.补肾温阳化瘀方对子宫内膜异位症肾阳虚血瘀证模型大鼠HIF-1α、PHD2、VHL表达的影响［J］. 中国中西医结合杂志, 2015, 35(10):1210-1217.
［22］邵进, 张岩, 王治,等.低氧诱导因子-1α参与骨发育及骨代谢调控的研究进展［J］.中国骨质疏松杂志, 2015, 6(3):349-355.
［23］涂福平, 黄莉, 王祥财. 缺氧环境下VHL和HIF-1α的变化［J］.医疗装备, 2017, 30(5):93-94.
［24］Liu Q, Tong D, Liu G, et al. HIF2Agermline-mutation-induced polycythemia in a patient with VHL-associated renal-cell carcinoma［J］. Cancer Biol Ther, 2017, 18(12):944-947.
［25］Liontos M, Trigka EA, Korkolopoulou P, et al. Expression and prognostic significance of VEGF and mTOR pathway proteins in metastatic renal cell carcinoma patients: a prognostic immunohistochemical profile for kidney cancer patients［J］. World J Urol, 2017, 35(3):1-9.
［26］Cho H, Du X, Rizzi J P, et al. On-Target efficacy of a HIF2α antagonist in preclinical kidney cancer models［J］. Nature, 2016, 539(7627):107-142.

[1]	范杰, 金永明, 江孝龙, 蒋国华. lncRNA HOTAIR调控miR-206影响类风湿关节炎滑膜细胞增殖和凋亡的分子机制研究[J]. 中国临床药理学与治疗学, 2023, 28(7): 736-742.
[2]	王安婧, 王亚亚, 梁轩, 闫雅婕, 苏静, 李彩东. 基于PPAR治疗胆汁淤积性肝病的机制与药物研究进展[J]. 中国临床药理学与治疗学, 2023, 28(7): 796-808.
[3]	王艳, 张金辉, 赵永辰, 刘宏祥, 刘亚维, 苗欢欢, 杨信才. 红杉黄酮通过下调PI3K/AKT信号通路抑制胃癌细胞增殖侵袭等干细胞特性[J]. 中国临床药理学与治疗学, 2023, 28(5): 508-513.
[4]	刘佳君, 阙祖俊, 田建辉. 金复康有效成分抑制肺癌循环肿瘤细胞募集中性粒细胞能力的比较研究[J]. 中国临床药理学与治疗学, 2023, 28(1): 1-9.
[5]	刘佳佳, 高岩, 张业慧, 武静. CRABP2在肿瘤发生发展中的作用研究[J]. 中国临床药理学与治疗学, 2023, 28(1): 86-92.
[6]	丁嘉敏, 王友娣, 赵夏丽, 姜根风, 陈思文, 洪程程, 李书勤, 胡卫华. 生长激素和维生素E联合应用治疗薄型子宫内膜的研究[J]. 中国临床药理学与治疗学, 2022, 27(8): 857-862.
[7]	眭玉霞, 庄捷, 庄敏, 黄桂. Delicaflavone通过PI3K/AKT/mTOR通路调控上皮-间质转化抑制吉非替尼耐药肺癌PC-9/GR细胞迁移和侵袭[J]. 中国临床药理学与治疗学, 2022, 27(6): 614-621.
[8]	王新丽, 许霞青, 方寒冰, 郭玉忠. 佛波酯增强顺铂的抗肿瘤活性及降低其肾毒性的作用研究[J]. 中国临床药理学与治疗学, 2022, 27(5): 535-543.
[9]	施经斌, 熊阳. 肿瘤微环境与乳腺癌细胞互作导致肿瘤转移的研究进展[J]. 中国临床药理学与治疗学, 2022, 27(5): 562-574.
[10]	李岩岩, 方骁杰, 尹鑫, 孙曦, 饶春晖. circZNF124通过靶向miR-4262调控结直肠癌SW620细胞增殖、迁移及侵袭的机制研究[J]. 中国临床药理学与治疗学, 2022, 27(11): 1231-1239.
[11]	雷静, 何平, 蔡文琳, 张永哲, 张蓓茹, 刘大军. 冬虫夏草提取物通过抑制PI3K/Akt通路抑制HBx诱导的系膜细胞增殖和细胞外基质沉积[J]. 中国临床药理学与治疗学, 2022, 27(11): 1247-1254.
[12]	郭胜杰, 管汐蕊, 曹文莉, 梁思成, 齐晓怡, 葛广波, 吕沐瀚. 尿苷二磷酸葡醛酸转移酶介导的胆汁酸代谢过程及其内源和外源影响因素研究进展[J]. 中国临床药理学与治疗学, 2022, 27(1): 102-107.
[13]	周敏敏, 余露, 韩超, 顾达民, 雷道赟. 人类胆固醇酯转移蛋白rs5882基因多态性与围术期神经认知功能紊乱的相关性[J]. 中国临床药理学与治疗学, 2021, 26(7): 768-774.
[14]	陈林俊, 陈文斌. 岩大戟内酯B通过阻断PI3K/Akt/NF-κB通路抑制结肠癌细胞的增殖和转移[J]. 中国临床药理学与治疗学, 2021, 26(3): 250-257.
[15]	赵嫄, 聂倩文, 尹江燕, 郇轩, 田梅香, 张正义. 赖氨酸乙酰化的动态调节在心脏疾病中的研究进展[J]. 中国临床药理学与治疗学, 2021, 26(10): 1186-1192.

氟尿嘧啶调控VHL/HIF信号通路抑制肾癌细胞增殖、侵袭与转移的研究

Fluorouracil regulates VHL/HIF signaling pathway and inhibits proliferation, invasion and metastasis of renal carcinoma cells

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价