Du Ruixia, Liu Zhe, Hou Xuedong, Fu Gongbi, An Ning, Wang Liping
Department of Otorhinolaryngology, Shengjing Hospital, China Medical University, Shenyang, Liaoning 110004, P.R. China.
Department of Pancreatic Surgery, First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
Mol Med Rep. 2016 Jun;13(6):5045-52. doi: 10.3892/mmr.2016.5204. Epub 2016 Apr 28.
Genistein and trichostatin A (TSA) are two chemotherapeutic compounds with antitumor effects in different types of cancer cell. However, the effects of genistein and TSA on the HEp‑2 laryngeal cancer cell line remain to be fully elucidated. In the present study, it was found that genistein and TSA inhibited cell growth and cell migration, and promoted apoptosis in the HEp‑2 laryngeal cancer cell line. The HEp‑2 cells were treated with genistein, TSA or the two compounds in combination. Cell proliferation and apoptosis were measured using an MTT assay, Annexin V/propidium iodide staining and a TUNEL assay. Cell invasion was determined using a Matrigel‑based Transwell assay. Western blotting was used to examine the activation of the Akt pathway and the expression levels of pro‑or anti‑apoptotic proteins. Treatment with either genistein or TSA alone mildly inhibited cell viability, growth and invasion, and induced the apoptosis of the laryngeal cancer cells, whereas more marked effects were observed in the cells treated with the combination of the two compounds. In addition, genistein reversed endothelial growth factor‑induced epithelial‑mesenchymal transition (EMT) in the HEp‑2 cells, the effect of which were was further increased by joint application with TSA. Treatment of the HEp‑2 cells with genistein and TSA led to a significant reduction in the phosphorylation of Akt and activation of its downstream target, and resulted in peroxisome proliferator‑activated receptor‑γ cleavage, increased expression of B cell lymphoma‑2 (Bcl‑2)‑associated X protein and reduced the expression of Bcl‑2. In conclusion, the present study demonstrated that, with the involvement of TSA, genistein exhibited substantial advantages in inhibiting laryngeal carcinoma cell growth, invasion and EMT, and induced apoptosis, compared with genistein treatment alone, which occurred through the regulation of Akt activation and the apoptotic pathway.
染料木黄酮和曲古抑菌素A(TSA)是两种对不同类型癌细胞具有抗肿瘤作用的化疗化合物。然而,染料木黄酮和TSA对人喉癌细胞系HEp-2的作用仍有待充分阐明。在本研究中,发现染料木黄酮和TSA抑制HEp-2人喉癌细胞系的细胞生长和细胞迁移,并促进其凋亡。用染料木黄酮、TSA或两种化合物联合处理HEp-2细胞。使用MTT法、膜联蛋白V/碘化丙啶染色和TUNEL法检测细胞增殖和凋亡。使用基于基质胶的Transwell法测定细胞侵袭。采用蛋白质免疫印迹法检测Akt信号通路的激活以及促凋亡或抗凋亡蛋白的表达水平。单独使用染料木黄酮或TSA处理可轻度抑制细胞活力、生长和侵袭,并诱导喉癌细胞凋亡,而在两种化合物联合处理的细胞中观察到更显著的效果。此外,染料木黄酮可逆转内皮生长因子诱导的HEp-2细胞上皮-间质转化(EMT),与TSA联合应用可进一步增强其效果。用染料木黄酮和TSA处理HEp-2细胞导致Akt磷酸化显著降低及其下游靶点激活,并导致过氧化物酶体增殖物激活受体γ裂解,B细胞淋巴瘤-2(Bcl-2)相关X蛋白表达增加,Bcl-2表达降低。总之,本研究表明,与单独使用染料木黄酮相比,在TSA的参与下,染料木黄酮在抑制喉癌细胞生长、侵袭和EMT以及诱导凋亡方面具有显著优势,这是通过调节Akt激活和凋亡途径实现的。