Wang Zhi-Ming, Yang Dong-Sheng, Liu Jie, Liu Hong-Bo, Ye Ming, Zhang Yu-Fei
Department of Oral and Maxillofacial Surgery, Shengjing Hospital of China Medical University, No. 36 Sanhao Street, Heping District, Shenyang, 110004, China.
Experimental Technology Center of China Medical University, No. 77 Puhe Road, Shenbeixin District, Shenyang, 110122, China.
Tumour Biol. 2016 Mar;37(3):3757-64. doi: 10.1007/s13277-015-4115-6. Epub 2015 Oct 14.
The objective of this study is to determine the effects of Rho-associated coiled-coil containing protein kinase (ROCK) inhibitor Y-27632 on the growth, invasion, and migration of Tca8113 and CAL-27 cells in tongue squamous cell carcinoma (TSCC). The methods of the study are as follows: After being routinely cultured for 24 h, Tca8113 and CAL-27 cells were treated with Y-27632 solution. The morphological change of Y-27632-treated cells was observed under an optical microscope and an inverted microscope; MTT assay was performed to measure the optical density (OD) of cells and calculate cell growth inhibition rate; the change of apoptosis was detected by AnnexinV-FITC/PI assay; cell invasion and migration were measured by Transwell assay. The results were as follows: (1) With increasing concentration of Y-27632, cell morphology changed and cell apoptosis appeared; (2) MTT assay showed that inhibition effect of Y-27632 on Tca8113 and CAL-27 cells was enhanced with increasing concentrations and time (all P < 0.01); (3) Apoptosis showed that, compared with controls, the number of apoptosis cells in experimental groups was significantly increased (all P < 0.01). Apoptosis rate was elevated with increasing concentrations of Y-27632; (4) Transwell assay showed, after a treatment with Y-27632, the number of migrated and invaded Tca8113 and CAL-27 cells in each group was statistically different (all P < 0.01); compared with controls, the number of migrated cell in groups treated with Y-27632 was decreased and less Tca8113 and CAL-27 cells in experimental groups passed through polycarbonate membrane (all P < 0.05). The study concludes that Y-27632 can inhibit the growth, invasion, and migration of Tca8113 and CAL-27 cells, suggesting that Y-27632 may be therapeutically useful in TSCC.
本研究旨在确定Rho相关卷曲螺旋蛋白激酶(ROCK)抑制剂Y-27632对舌鳞状细胞癌(TSCC)中Tca8113和CAL-27细胞生长、侵袭及迁移的影响。研究方法如下:Tca8113和CAL-27细胞常规培养24小时后,用Y-27632溶液处理。在光学显微镜和倒置显微镜下观察经Y-27632处理的细胞形态变化;采用MTT法检测细胞光密度(OD)并计算细胞生长抑制率;通过AnnexinV-FITC/PI法检测细胞凋亡变化;采用Transwell法检测细胞侵袭和迁移能力。结果如下:(1)随着Y-27632浓度增加,细胞形态改变并出现细胞凋亡;(2)MTT法显示,Y-27632对Tca8113和CAL-27细胞的抑制作用随浓度和时间增加而增强(所有P<0.01);(3)凋亡检测显示,与对照组相比,实验组凋亡细胞数量显著增加(所有P<0.01)。凋亡率随Y-27632浓度增加而升高;(4)Transwell法显示,经Y-27632处理后,各组Tca8113和CAL-27细胞迁移和侵袭数量差异有统计学意义(所有P<0.01);与对照组相比,Y-27632处理组迁移细胞数量减少,实验组中穿过聚碳酸酯膜的Tca8113和CAL-27细胞减少(所有P<0.05)。研究得出结论,Y-27632可抑制Tca8113和CAL-27细胞的生长、侵袭及迁移,提示Y-27632可能对TSCC具有治疗作用。