TH287对口腔鳞状细胞癌CAL27细胞增殖和迁移的抑制作用
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摘要
目的:TH287作为MutT同系物1(MutT homolog1,MTH1)蛋白的抑制剂,可有效抑制癌细胞中过表达的MTH1蛋白,从而解除对下游凋亡相关蛋白半胱氨酰天冬氨酸特异性蛋白酶3 (cysteinyl aspartate-specific protease-3,Caspase-3)介导细胞凋亡的抑制作用。本研究旨在检测TH287对口腔鳞状细胞癌CAL27细胞增殖性和迁移性的抑制作用。方法:通过噻唑蓝(methyl thiazol tetrazolium,MTT)法检测TH287对口腔鳞状细胞癌CAL27细胞增殖性的抑制作用并确定药物最适浓度;通过流式细胞术检测TH287对CAL27细胞凋亡的促进作用;通过Western blot实验检测TH287对MTH1蛋白表达的抑制作用;通过划痕实验和Transwell迁移实验检测TH287对CAL27细胞迁移性的抑制作用。结果:TH287有效地抑制了CAL27细胞的增殖性,其最适浓度为100μmol/L。TH287同时也促进CAL27细胞发生凋亡并抑制MTH1蛋白的表达。除此之外,CAL27细胞的迁移性也受到了抑制。结论:TH287可以抑制CAL27细胞的增殖并促进其凋亡。与此同时,CAL27细胞的迁移性也明显受到抑制。TH287对CAL27细胞增殖性和迁移性的抑制作用可能与MTH1蛋白表达受到抑制有关。因此,TH287可能成为口腔鳞状细胞癌靶点治疗的新药物。
Objective: To test the inhibition of TH287 on the proliferation and metastasis of CAL27. Methods: MTT assay was used to test the inhibition of TH287 on the proliferation of CAL27 and ascertain the appropriate concentration. The promotion of TH287 on the apoptosis of CAL27 was tested by the flow cytometry. The inhibition of TH287 on the MTH1 expression was examined by the western blot. The inhibition of TH287 on the metastasis of CAL27 was examined by the scratch test and Transwell(migration). Results: TH287 could inhibit the proliferation of CAL27 effectively and the appropriate concentration was 100 μM. TH287 could also promote the apoptosis of CAL27 and inhibit the MTH1 expression. In addition, the metastasis of CAL27 was also inhibited by the TH287. Conclusion: TH287 could inhibit the proliferation and promote the apoptosis of CAL27. Meanwhile, the metastasis of CAL27 was also inhibited by TH287. The inhibition of TH287 on the proliferation and metastasis of CAL27 may be caused by the suppression of MTH1 expression. Therefore, TH287 has great potential to be a new drug in targeted therapy.
Objective: To test the inhibition of TH287 on the proliferation and metastasis of CAL27. Methods: MTT assay was used to test the inhibition of TH287 on the proliferation of CAL27 and ascertain the appropriate concentration. The promotion of TH287 on the apoptosis of CAL27 was tested by the flow cytometry. The inhibition of TH287 on the MTH1 expression was examined by the western blot. The inhibition of TH287 on the metastasis of CAL27 was examined by the scratch test and Transwell(migration). Results: TH287 could inhibit the proliferation of CAL27 effectively and the appropriate concentration was 100 μM. TH287 could also promote the apoptosis of CAL27 and inhibit the MTH1 expression. In addition, the metastasis of CAL27 was also inhibited by the TH287. Conclusion: TH287 could inhibit the proliferation and promote the apoptosis of CAL27. Meanwhile, the metastasis of CAL27 was also inhibited by TH287. The inhibition of TH287 on the proliferation and metastasis of CAL27 may be caused by the suppression of MTH1 expression. Therefore, TH287 has great potential to be a new drug in targeted therapy.
引文
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[3] Xu R,Zeng G,Gao J,et al.miR-138 suppresses the proliferation of oral squamous cell carcinoma cells by targeting Yes-associated protein 1 [J].Oncol Rep,2015,34(4):2171-2178.
[4] 韩影,苏彤,林璐,等.口腔癌对侧颈淋巴结转移的临床研究[J].口腔医学研究,2016,32(2):154-157.
[5] Cheng KC,Cahill DS,Kasai H,et al.8-Hydroxyguanine,an abundant form of oxidative DNA damage,causes G-T and A-C substitutions [J].J Biol Chem,1992,267(1):166-172.
[6] Leiros I,Nabong MP,Grosvik K,et al.Structural basis for enzymatic excision of N1-methyladenine and N3-methylcytosine from DNA [J].EMBO J,2014,26(8):2206-2217.
[7] Bowerman B.Cell biology.Oxidative stress and cancer:a beta-catenin convergence [J].Science,2005,308(5725):1119-1120.
[8] Gad H,Koolmeister T,Jemth AS,et al.MTH1 inhibition eradicates cancer by preventing sanitation of the dNTP pool [J].Nature,2014,508(7495):215-221.
[9] Nakabeppu Y,Oka S,Sheng Z,et al.Programmed cell death triggered by nucleotide pool damage and its prevention by MutT homolog-1 (MTH1) with oxidized purine nucleoside triphosphatase [J].Mutat Res,2010,703(1):51-58.
[10] Nakabeppu Y.Cellular levels of 8-oxoguanine in either DNA or the nucleotide pool play pivotal roles in carcinogenesis and survival of cancer cells [J].Int J Mol Sci,2014,15(7):12543-12557.
[11] Brozmanová J,Dudás A,Henriques JA.Repair of oxidative DNA damage--an important factor reducing cancer risk.Minireview [J].Neoplasma,2001,48(2):85-93.
[12] Sakai Y,Furuichi M,Takahashi M,et al.A molecular basis for the selective recognition of 2-hydroxy--dATP and 8-oxo-dGTP by human MTH1 [J].J Biol Chem,2002,277(10):8579-8587.
[13] Nakabeppu Y.Molecular genetics and structural biology of human MutT homolog,MTH1 [J].Mutat Res,2001,477(1-2):59-70.
[14] Qing X,Shao Z,Lv X,et al.Anticancer effect of (S)-crizotinib on osteosarcoma cells by targeting MTH1 and activating reactive oxygen species [J].Anticancer Drugs,2018,29(4):341-352.
[15] Abbas HHK,Alhamoudi KMH,Evans MD,et al.MTH1 deficiency selectively increases non-cytotoxic oxidative DNA damage in lung cancer cells:more bad news than good [J].BMC Cancer,2018,18(1):423.
[16] Tamura T,Ichikawa T,Nakahata S,et al.Loss of NDRG2 expression confers oral squamous cell carcinoma with enhanced metastatic potential [J].Cancer Res,2017,77(9):2363-2374.
[17] 陈震,郭燕.E-cadherin和Vimentin在OSCC上皮-间质转化中的作用及表达[J].口腔医学研究,2016,32(8):857-860.
[18] Joseph JP,Harishankar MK,Pillai AA,et al.Hypoxia induced EMT:A review on the mechanism of tumor progression and metastasis in OSCC [J].Oral Oncol,2018,80:23-32.
[19] Hong KO,Kim JH,Hong JS,et al.Inhibition of Akt activity induces the mesenchymal-to-epithelial reverting transition with restoring E-cadherin expression in KB and KOSCC-25B oral squamous cell carcinoma cells [J].J Exp Clin Cancer Res,2009,28(1):28.
[20] Li J,Yang CC,Tian XY,et al.MutT-related proteins are novel progression and prognostic markers for colorectal cancer [J].Oncotarget,2017,8(62):105714-105726.
[21] Giribaldi MG,Munoz A,Halvorsen K,et al.MTH1 expression is required for effective transformation by oncogenic HRAS [J].Oncotarget,2015,6(13):11519-11529.