PFKFB3 promotes cell proliferation, migration, invasion and metastasis in adenoid cystic carcinoma ACC-2 cells

Abstract

Abstract:Objective To evaluate the expression of 6-phosphofructo-2kinase/fructose-2, 6-bisphosphatase 3 (PFKFB3) in adenoid cystic carcinoma and the effects of PFKFB3 on the proliferation, migration, invasion and tumorigenesis in vivo of adenoid cystic carcinoma cells. Methods Immunohistochemistry and Western blotting were applied to detect the expression of PFKFB3 in 29 cases of adenoid cystic carcinoma tissues and 10 cases of paracarcinoma normal salivary gland tissues. PFKFB3 in adenoid cystic carcinoma ACC2 cells was silenced by adenovirus vector. The effects of PFKFB3 silence on cell proliferation, migration, invasion and tumorigenesis in vivo and distant metastasis of adenoid cystic carcinoma cells were evaluated by cell proliferation assay, wound healing assay, Transwell assay, xenografted mice model and lung metastasis mice model. Results The results of immunohistochemical staining showed that the positive expression rate of PFKFB3 in adenoid cystic carcinoma tissues was 93.1% (27/29), and the positive expression rate of PFKFB3 in normal salivary gland tissues was 20.0% (2/10), with a significant difference (χ2=20.84, P＜0.001). The results of methyl thiazolyl tetrazolium (MTT) assay showed that, compared with ACC2 group, the proliferation activity of cancer cells with silence of PFKFB3 (PFKFB3-ACC-2) was significantly suppressed for 72 h (1.8±0.2 vs. 4.7±0.8, t=3.582, P=0.001). The results of wound healing assay showed that after scraping cells away for 24 h, the number of cells in the scratch area was 99.8±13.2 in the PFKFB3-ACC2 group, which was significantly less than that in the ACC-2 group (263.0±97.4, t=2.868, P=0.029). Transwell results indicated that the number of cells passing through matrigel was 17.6±2.1 in the PFKFB3-ACC-2 group, which was less than that in the ACC-2 group (28.6±3.8, t=4.452, P=0.004). The tumor volume in the PFKFB3-ACC-2 ［(623.5±134.1) mm3］ was smaller than that in the ACC-2 group ［(1 621.0±278.1) mm3, t=4.213, P=0.001］. More pulmonary metastases were found in the ACC-2 group than PFKFB3-ACC-2 group (18.1±3.2 vs. 4.1±2.2, t=6.322, P=0.001). Conclusion The expression of PFKFB3 is higher in adenoid cystic carcinoma tissue than normal salivary gland tissue, and the highly expressed PFKFB3 plays a driving role in the proliferation, migration, invasion and metastasis in adenoid cystic carcinoma.

Key words:Carcinoma, adenoid cystic,Glycolysis,PFKFB3,Tumor development

Qiao Yan, Liu Xin. PFKFB3 promotes cell proliferation, migration, invasion and metastasis in adenoid cystic carcinoma ACC-2 cells[J]. Journal of International Oncology, 2018, 45(10): 577-582.

References

［1］ Dillon PM, Chakraborty S, Moskaluk CA, et al. Adenoid cystic carcinoma: a review of recent advances, molecular targets and clinical trials［J］. Head Neck, 2016, 38 (4): 620-627. DOI: 10.1002/hed.23925. ［2］黄擎, 韩楠男, 刘胜文, 等. 唾液腺腺样囊性癌颈淋巴结转移规律的临床研究［J］. 中国肿瘤临床, 2016, 43(24): 1094-1098. DOI: 10.3969/j.issn.10008179.2016.24.125. ［3］严斐, 王超, 黎婷, 等. miR21对唾液腺腺样囊性癌细胞增殖和凋亡的影响及机制研究［J］. 中国口腔颌面外科杂志, 2017, 15(3): 209-213. DOI: 10.19438/j.cjoms.2017.03.004. ［4］ Chen W, Cao G, Yuan X, et al. Notch1 knockdown suppresses proliferation, migration and metastasis of salivary adenoid cystic carcinoma cells［J］. J Transl Med, 2015, 13: 167. DOI: 10.1186/s12967-015-0520-2. ［5］梁军, 黄志权, 游云华, 等. HIF1α和VEGF的表达与腺样囊性癌临床病理的相关性研究［J］. 中国口腔颌面外科杂志, 2010, 8(6): 548-554. ［6］ Ali S, Yeo JC, Magos T, et al. Clinical outcomes of adenoid cystic carcinoma of the head and neck: a single institution 20year experience［J］. J Laryngol Otol, 2016, 130 (7): 680-685. DOI: 10.1017/S0022215116008124. ［7］黄应斌, 农晓琳, 杨亦萍, 等. CXCL12/CXCR4在腺样囊性癌中的表达及对预后的价值［J］. 实用口腔医学杂志, 2015, 31(2): 210-214. DOI: 10.3969/j.issn.1001-3733.2015.02.014. ［8］周宇会, 魏九峰, 李国东, 等. Warburg效应在肿瘤中的研究进展［J］. 国际肿瘤学杂志, 2017, 44(10): 762-766. DOI: 10.3760/cma.j.issn.1673422X.2017.10.010. ［9］王豪华, 陈秀星, 郭桂芳. 己糖激酶Ⅱ与肿瘤增殖及靶向治疗［J］. 国际肿瘤学杂志, 2017, 44(3): 213-216. DOI: 10.3760/cma.j.issn.1673422X.2017.03.014. ［10］ Douglas JG, Laramore GE, AustinSeymour M, et al. Treatment of locally advanced adenoid cystic carcinoma of the head and neck with neutron radiotherapy［J］. Int J Radiat Oncol Biol Phys, 2000, 46(3): 551-557. ［11］ Jang S, Patel PN, Kimple RJ, et al. Clinical outcomes and prognostic factors of adenoid cystic carcinoma of the head and neck［J］. Anticancer Res, 2017, 37(6): 3045-3052. DOI: 10.21873/anticanres.11659. ［12］ Bell D, Hanna EY. Head and neck adenoid cystic carcinoma: what is new in biological markers and treatment?［J］. Curr Opin Otolaryngol Head Neck Surg, 2013, 21(2): 124-129. DOI: 10.1097/MOO.0b013e32835c05fd. ［13］ Oldham RK. Cancer biotherapy: more than immunotherapy［J］. Cancer Biother Radiopharm, 2017, 32(4): 111114. DOI: 10.1089/cbr.2017.28999.old. ［14］ Neugent ML, Goodwin J, Sankaranarayanan I, et al. A new perspective on the heterogeneity of cancer glycolysis［J］. Biomol Ther (Seoul), 2018, 26(1): 10-18. DOI: 10.4062/biomolther.2017.210. ［15］ Li HM, Yang JG, Liu ZJ, et al. Blockage of glycolysis by targeting PFKFB3 suppresses tumor growth and metastasis in head and neck squamous cell carcinoma［J］. J Exp Clin Cancer Res, 2017, 36(1): 7. DOI: 10.1186/s13046-016-0481-1. ［16］ Chen L, Zhao J, Tang Q, et al. PFKFB3 control of cancer growth by responding to circadian clock outputs［J］. Sci Rep, 2016, 6: 24324. DOI: 10.1038/srep24324. ［17］ Gu M, Li L, Zhang Z, et al. PFKFB3 promotes proliferation, migration and angiogenesis in nasopharyngeal carcinoma［J］. J Cancer, 2017, 8(18): 38873896. DOI: 10.7150/jca.19112. ［18］ Peng F, Li Q, Sun JY, et al. PFKFB3 is involved in breast cancer proliferation, migration, invasion and angiogenesis［J］. Int J Oncol, 2018, 52(3): 945-954. DOI: 10.3892/ijo.2018.4257. ［19］ Clem BF, O′Neal J, Tapolsky G, et al. Targeting 6phosphofructo2kinase (PFKFB3) as a therapeutic strategy against cancer［J］. Mol Cancer Ther, 2013, 12(8): 1461-1470. DOI: 10.1158/1535-7163.MCT-13-0097. ［20］ Telang S, Clem BF, Klarer AC, et al. Small molecule inhibition of 6phosphofructo2kinase suppresses T cell activation［J］. J Transl Med, 2012, 10: 95. DOI: 10.1186/1479-5876-10-95.

[1]	Feng Dongxu, Wu Wei, Gao Pingfa, Wang Jun, Shi Lijuan, Chen Dawei, Li Wenbing, Zhang Meifeng.Effects of miR-451 on glycolysis and apoptosis of breast cancer cells by regulating Rho/ROCK1 pathway[J]. Journal of International Oncology, 2023, 50(8): 449-456.
[2]	Zhong Chenyi, Mao Yundong.Regulating effect of lncRNA on aerobic glycolysis in tumor cells[J]. Journal of International Oncology, 2019, 46(3): 170-173.
[3]	Zhou Yuhui, Wei Jiufeng, Li Guodong, Liu Ming.Warburg effect on tumor[J]. Journal of International Oncology, 2017, 44(10): 762-766.
[4]	Wang Juan, Cheng Bin.Role of M2type pyruvate kinase in the metabolism of tumor[J]. Journal of International Oncology, 2014, 41(1): 17-20.
[5]	SUN Jie, MENG Xiang-Jun.Abnormal glycometabolism in tumor cells[J]. Journal of International Oncology, 2013, 40(12): 883-885.
[6]	PANG Yu-Yang, WANG Ting, CHEN Fang-Yuan.Glycolysis abnormality in neoplasm cells and the targeted therapy[J]. Journal of International Oncology, 2012, 39(10): 726-728.