毛蕊花苷通过上调Max蛋白抑制口腔鳞癌细胞的迁移和侵袭
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摘要
目的:探讨毛蕊花苷对口腔鳞癌细胞迁移和侵袭的影响及其机制。方法:质谱分析空白对照组及毛蕊花苷处理组口腔鳞癌细胞HN4中蛋白的表达谱,筛选毛蕊花苷处理后丰度高且表达显著增加的蛋白,用Western blot验证质谱分析的结果,并在HN4细胞中运用shRNA干扰关键蛋白的表达,分析毛蕊花苷处理对HN4细胞迁移和侵袭的影响。结果:质谱分析结果显示经毛蕊花苷处理后,HN4细胞中Max、TMPRSS11F、ROBO4和AP4E1等18种蛋白表达显著升高(变化倍数>1.5),对其中改变最明显的Max蛋白进行Western blot检测,结果与质谱芯片结果一致;shRNA干扰HN4细胞中Max的表达,qRT-PCR和Western blot检测发现HN4细胞中Max的表达明显下调,运用毛蕊花苷刺激HN4细胞,发现毛蕊花苷可显著抑制HN4细胞的迁移和侵袭。然而,当敲减Max蛋白后,毛蕊花苷抑制HN4细胞迁移和侵袭的能力显著减弱。结论:毛蕊花苷可能通过增强口腔鳞癌细胞中Max的表达抑制口腔鳞癌细胞的迁移和侵袭。
Objective:To analyze the effect and mechanism of verbascoside on the migration and invasion of oral squamous cell carcinoma. Methods:The expression profiles of HN4 protein in oral squamous cell carcinoma cells treated with verbascoside and the blank control group were analyzed by mass spectrometry. Proteins with high abundance and significantly increased expression after verbascoside treatment were screened. The results of mass spectrometry were verified by Western blotting. The expression of key proteins in HN4 cells was interfered by shRNA to analyze verbascoside. Effects of verbascoside on migration and invasion of squamous cell carcinoma HN4 cells were analyzed. Results:The results of mass spectrometry analysis showed that 18 proteins,including Max,MPRSS11 F,ROBO4,AP4 E1,and etc,was significantly increased in oral squamous cell carcinoma HN4 cells after treatment with verbascoside(fold change>1.5). The Max proteins with the most obvious changes were detected by Western blotting. The results of Western blotting were consistent with those of mass spectrometry microarray. shRNA interfered with the expression of Max in oral squamous cell carcinoma HN4 cells. qRT-PCR and Western blotting showed that the expression of Max in HN4 cells was significantly down-regulated. Further stimulation of HN4 cells with verbascoside showed that the migration and invasion of HN4 cells were significantly inhibited. However,the ability of verbascoside to inhibit the migration and invasion of HN4 cells decreased significantly after knockdown of Max protein. Conclusion:It is possible that verbascoside inhibits the migration and invasion of oral squamous cell carcinoma cells by enhancing the expression of Max in oral squamous cell carcinoma cells.
Objective:To analyze the effect and mechanism of verbascoside on the migration and invasion of oral squamous cell carcinoma. Methods:The expression profiles of HN4 protein in oral squamous cell carcinoma cells treated with verbascoside and the blank control group were analyzed by mass spectrometry. Proteins with high abundance and significantly increased expression after verbascoside treatment were screened. The results of mass spectrometry were verified by Western blotting. The expression of key proteins in HN4 cells was interfered by shRNA to analyze verbascoside. Effects of verbascoside on migration and invasion of squamous cell carcinoma HN4 cells were analyzed. Results:The results of mass spectrometry analysis showed that 18 proteins,including Max,MPRSS11 F,ROBO4,AP4 E1,and etc,was significantly increased in oral squamous cell carcinoma HN4 cells after treatment with verbascoside(fold change>1.5). The Max proteins with the most obvious changes were detected by Western blotting. The results of Western blotting were consistent with those of mass spectrometry microarray. shRNA interfered with the expression of Max in oral squamous cell carcinoma HN4 cells. qRT-PCR and Western blotting showed that the expression of Max in HN4 cells was significantly down-regulated. Further stimulation of HN4 cells with verbascoside showed that the migration and invasion of HN4 cells were significantly inhibited. However,the ability of verbascoside to inhibit the migration and invasion of HN4 cells decreased significantly after knockdown of Max protein. Conclusion:It is possible that verbascoside inhibits the migration and invasion of oral squamous cell carcinoma cells by enhancing the expression of Max in oral squamous cell carcinoma cells.
引文
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[12] Iqbal MS,Chaw C,Kovarik J,et al. Primary concurrent chemoradiation in head and neck cancers with weekly cisplatin chemotherapy:analysis of compliance,toxicity and survival[J]. Int Arch Otorhinolaryngol,2017,21(2):171-177
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[14]Yabuuchi H,Kuroiwa T,Tajima T,et al. Efficacy of intraarterial infusion therapy using a combination of cisplatin and docetaxel for recurrent head and neck cancers compared with cisplatin alone[J]. Clin Oncol(R Coll Radiol),2003,15(8):467-472
[15]彭素萍,巩珺,苗瑞娟,等.连翘酯苷B与毛蕊花苷稳定性研究[J].今日药学,2012,22(6):236-238
[16] Ferré-D’AmaréAR,Prendergast GC,Edward B,et al.Recognition by Max of its cognate DNA through a dimeric b/HLH/Z domain[J]. Nature,1993,363(6424):38-45
[2]赵莉琳,刘阳. PCNA对口腔鳞癌细胞凋亡和增殖的影响[J].解放军医学杂志,2014,39(10):795-799
[3] da Silva SD,Ferlito A,Takes RP,et al. Advances and applications of oral cancer basic research[J]. Oral Oncol,2011,47(9):783-791
[4]郑帅,杨敏.毛蕊花糖苷治疗糖尿病肾病的研究进展[J].医学综述,2018,24(16):3232-3241
[5]邓锦芳,熊英,王淳,等.毛蕊花糖苷对顺铂诱导小鼠卵巢损害的保护作用[J].解剖科学进展,2017,23(3):257-261
[6]邓海峰,孙缦利,陈浩,等.毛蕊花糖苷对抑郁症大鼠行为学和前额叶皮层内质网应激的影响[J].中国病理生理杂志,2018,34(1):101-106
[7]高莉,彭晓明,霍仕霞,等.毛蕊花糖苷改善D-半乳糖致亚急性衰老小鼠脑损伤的作用[J].中草药,2014,45(1):81-85
[8]宋小敏,廖理曦,董馨,等.毛蕊花糖苷抑制脂多糖诱导的BV-2小胶质细胞炎症反应及机制研究[J].中国中药杂志,2016,41(13):2506-2510
[9] Alipieva K,Korkina L,Orhan IE,et al. Verbascoside:a review of its occurrence,(bio)synthesis and pharmacological significance[J]. Biotechnol Adv,2014,32(6):1065-1076
[10] Zhang YQ,Yuan Y,Wu HM,et al. Effect of verbascoside on apoptosis and metastasis in human oral squamous cell carcinoma[J]. Int J Cancer,2018,143(4):980-991
[11] Petruzzi MN,Cherubini K,Salum FG,et al. Role of tumour-associated macrophages in oral squamous cells carcinoma progression:an update on current knowledge[J]. Diagn Pathol,2017,12(1):32
[12] Iqbal MS,Chaw C,Kovarik J,et al. Primary concurrent chemoradiation in head and neck cancers with weekly cisplatin chemotherapy:analysis of compliance,toxicity and survival[J]. Int Arch Otorhinolaryngol,2017,21(2):171-177
[13]Hayashi Y,Mitsudo K,Sakuma K,et al. Clinical outcomes of retrograde intra-arterial chemotherapy concurrent with radiotherapy for elderly oral squamous cell carcinoma patients aged over 80 years old[J]. Radiat Oncol,2017,12(1):112
[14]Yabuuchi H,Kuroiwa T,Tajima T,et al. Efficacy of intraarterial infusion therapy using a combination of cisplatin and docetaxel for recurrent head and neck cancers compared with cisplatin alone[J]. Clin Oncol(R Coll Radiol),2003,15(8):467-472
[15]彭素萍,巩珺,苗瑞娟,等.连翘酯苷B与毛蕊花苷稳定性研究[J].今日药学,2012,22(6):236-238
[16] Ferré-D’AmaréAR,Prendergast GC,Edward B,et al.Recognition by Max of its cognate DNA through a dimeric b/HLH/Z domain[J]. Nature,1993,363(6424):38-45