四氢生物蝶呤对胰腺癌细胞增殖、迁移及凋亡的影响

doi:10.13418/j.issn.1001-165x.2023.6.13

摘要/Abstract

摘要： 目的探究四氢生物蝶呤（tetrahydrobiopterin，BH4）对胰腺癌细胞生物学行为的影响及可能机制。方法细胞分为对照组与实验组，实验组根据BH4浓度（2、4、8、12 μM）分为4组，采用CCK-8、Hoechst及划痕实验检测细胞的增殖、凋亡和迁移能力，RT-qPCR检测细胞中Notch1 mRNA表达情况，Western blotting检测细胞中E-Cadherin与N-cadherin表达情况。结果与对照组相比较，随着BH4浓度的增加，HPAC与BxPC-3细胞的活性与迁移能力逐渐下降，细胞凋亡率逐渐增高，且8 μM、12 μM浓度的BH4作用有显著意义（P<0.01，P<0.05）；与对照组相比较，12 μM BH4组细胞中Notch1 mRNA显著降低（P<0.05），E-cadherin蛋白表达水平升高（P<0.01，P<0.05），N-cadherin蛋白表达水平降低（P<0.05）。结论 BH4可抑制胰腺癌细胞的增殖和迁移，促进其凋亡，其机制可能与抑制上皮细胞-间充质转化（epithelial-mesenchymal transition，EMT）有关，而Notch1在EMT发生过程中改变，提示Notch1信号通路可能参与该过程的调控。

关键词: 四氢生物蝶呤, , , 胰腺癌, , , 上皮间质转化, , , Notch信号通路

Abstract: Objective To investigate the effect of tetrahydrobiopterin (BH4) on the biological behavior of pancreatic cancer cells and its possible mechanism. Methods The cells were divided into a control group and an experimental group. The experimental group was divided into 4 subgroups according to the concentration of BH4 (2 μM, 4 μM, 8 μM, 12 μM BH4 groups). The ability of cell proliferation, apoptosis and migration were detected by CCK-8 assay, Hoechst assay and scratch assay. The expression of Notch1 mRNA was detected by RT-qPCR and the expressions of E-cadherin and N-cadherin were detected by Western blotting. Results Compared with the control group, with the increasing of BH4 concentration, the activity and migration ability of HPAC and Bxpc-3 cell decreased and the apoptosis rate increased, and the effect of BH4 was significant at 8 μM and 12 μM (P<0.01，P<0.05). Compared with the control group, Notch1 mRNA was significantly decreased in experimental group (BH4 12 μM) (P<0.05), the expression level of E-cadherin increased (P<0.01, P<0.05) , while that of N-cadherin decreased (P<0.05). Conclusions BH4 can inhibit the proliferation, migration and promote the apoptosis of pancreatic cancer cells. The mechanism may be related to the inhibition of epithelial-mesenchymal transition (EMT) process, while Notch1 changes during EMT. It suggests that the Notch1 signaling pathway may be involved in the regulation of this process.

Key words: , BH4, , , , Pancreatic cancer, , , Epithelial-mesenchymal transition, , , Notch signaling pathway

中图分类号:

R361.3

马文婷, 张晨, 谢青. 四氢生物蝶呤对胰腺癌细胞增殖、迁移及凋亡的影响[J]. 中国临床解剖学杂志, 2023, 41(6): 704-708.

Ma Wenting, Zhang Chen, Xie Qing. Effects of BH4 on proliferation, migration and apoptosis of pancreatic cancer cells[J]. Chinese Journal of Clinical Anatomy, 2023, 41(6): 704-708.

参考文献 21

[1]	Siegel RL, Miller KD, Fuchs HE, et al. Cancer statistics, 2021[J]. CA Cancer J Clin, 2021, 71(1): 7-33. DOI: 10.3322/caac.21654.
[2]	Gupta R, Amanam I, Chung V. Current and future therapies for advanced pancreatic cancer[J]. J Surg Oncol, 2017. 116(1): 25-34. DOI: 10.1002/jso.24623.
[3]	Christenson ES, Jaffee E, Azad NS. Current and emerging therapies for patients with advanced pancreatic ductal adenocarcinoma: a bright future[J]. Lancet Oncol, 2020, 21(3): e135-e145. DOI: 10.1016/S1470-2045(19)30795-8.
[4]	Moens AL, Kietadisorn R, Lin JY, et al. Targeting endothelial and myocardial dysfunction with tetrahydrobiopterin[J]. J Mol Cell Cardiol, 2011, 51(4): 559-563. DOI: 10.1016/j.yjmcc.2011.03.009.
[5]	McNeill E, Channon KM. The role of tetrahydrobiopterin in inflammation and cardiovascular disease[J]. Thromb Haemost, 2012, 108(5): 832-839. DOI: 10.1160/TH12-06-0424.
[6]	Bailey JD, Diotallevi M, Nicol T, et al. Nitric oxide modulates metabolic remodeling in inflammatory macrophages through TCA cycle regulation and itaconate accumulation[J]. Cell Rep, 2019, 28(1): 218-230. DOI: 10.1016/j.celrep.2019.06.018.
[7]	Kotamraju S, Williams CL, Kalyanaraman B. Statin-induced breast cancer cell death: role of inducible nitric oxide and arginase-dependent pathways[J]. Cancer Res, 2007, 67(15): 7386-7394. DOI: 10.1158/0008-5472.CAN-07-0993.
[8]	Zheng X, Fernando V, Sharma V, et al. Correction of arginine metabolism with sepiapterin-the precursor of nitric oxide synthase cofactor BH(4)-induces immunostimulatory-shift of breast cancer[J]. Biochem Pharmacol, 2020,176:113887. DOI: 10.1016/j.bcp. 2020. 113887.
[9]	Cho YR, Kim SH, Ko HY, et al. Sepiapterin inhibits cell proliferation and migration of ovarian cancer cells via down-regulation of p70S6K-dependent VEGFR-2 expression[J]. Oncol Rep, 2011, 26(4): 861-867. DOI: 10.3892/or.2011.1335.
[10]	Gonçalves DA, Xisto R, Gonçalves JD, et al. Imbalance between nitric oxide and superoxide anion induced by uncoupled nitric oxide synthase contributes to human melanoma development[J]. Int J Biochem Cell Biol,2019, 115:105592. DOI: 10.1016/j.biocel. 2019. 105592.
[11]	Chen H, Zhuo Q, Ye Z, et al. Organoid model: a new hope for pancreatic cancer treatment[J]. Biochim Biophys Acta Rev Cancer, 2021, 1875(1): 188466. DOI: 10.1016/j.bbcan.2020.188466.
[12]	Zhao Z, Liu W. Pancreatic cancer: a review of risk factors, diagnosis, and treatment[J]. Technol Cancer Res Treat, 2020, 19: 1533033820962 117. DOI: 10.1177/1533033820962117.
[13]	Zhong GC, Zhao ZB, Cheng Y, et al. Epigenetic silencing of GCH1promotes hepatocellular carcinoma growth by activating superoxide anion-mediated ASK1/p38 signaling via inhibiting tetrahydrobiopterin de novo biosynthesis[J]. Free Radic Biol Med, 2021, 168: 81-94. DOI: 10.1016/j.freeradbiomed.2021.03.025.
[14]	Cronin S, Seehus C, Weidinger A, et al. The metabolite BH4 controls T cell proliferation in autoimmunity and cancer[J]. Nature, 2018, 563(7732): 564-568. DOI: 10.1038/s41586-018-0701-2.
[15]	Cao ZQ, Wang Z, Leng P. Aberrant N-cadherin expression in cancer[J]. Biomed Pharmacother,2019, 118:109320.DOI:10.1016/j.biopha. 2019. 109320.
[16]	Sanz Ressel BL, Massone AR, Barbeito CG. Aberrant expression of E-cadherin/β-catenin during epidermal tumourigenesis in dogs[J]. J Comp Pathol, 2020, 176: 1-9. DOI: 10.1016/j.jcpa.2020.01.006.
[17]	Jang NR, Choi JH, Gu MJ. Aberrant expression of E-cadherin, N-cadherin, and P-cadherin in clear cell renal cell carcinoma: association with adverse clinicopathologic factors and poor prognosis[J]. Appl Immunohistochem Mol Morphol, 2021, 29(3): 223-230. DOI: 10.1097/PAI.0000000000000861.
[18]	Leong KG, Karsan A. Recent insights into the role of Notch signaling in tumorigenesis[J]. Blood, 2006, 107(6): 2223-2233. DOI: 10.1182/blood-2005-08-3329.
[19]	Chu Q, Wang L, Zhang J, et al. CDK5 positively regulates Notch1 signaling in pancreatic cancer cells by phosphorylation[J].Cancer Med, 2021, 10(11): 3689-3699. DOI: 10.1002/cam4.3916.
[20]	Feng L, Wang K, Tang P, et al. Deubiquitinase USP18 promotes the progression of pancreatic cancer via enhancing the Notch1-c-Myc axis[J]. Aging (Albany NY), 2020, 12(19): 19273-19292. DOI: 10.18632/aging.103760.
[21]	Lin J, Xu Z, Xie J, et al. Oncogene APOL1 promotes proliferation and inhibits apoptosis via activating NOTCH1 signaling pathway in pancreatic cancer[J]. Cell Death Dis, 2021, 12(8): 760. DOI: 10.1038/s41419-021-03985-1.