Jia Bo, Qiu Xiao-Ling, Chu Hong-Xing, Sun Xiang, Pan Jie, Wang Zhi-Ping, Zhao Jian-Jiang
Department of Oral Surgery, Guangdong Provincial Stomatological Hospital/Stomatological Hospital of Southern Medical University, Guangzhou 510280, China. E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2017 Aug 20;37(8):1104-1109. doi: 10.3969/j.issn.1673-4254.2017.08.17.
To detect CCR5 protein expression in different human tongue squamous cell carcinoma cells and observe the effect of macrophage inflammatory protein-1β (MIP-1β) on the proliferation and apoptosis of CAL-27 cells.
Western blotting and immunofluorescence staining were used to detect the expression of the CCR5, the receptor of MIP-1β, in 3 human tongue squamous cell carcinoma cells UM-1, CAL-27, and Tca-8113. CCK-8 assay was used to assess the proliferation of CAL-27 cells stimulated with 10, 20, and 40 ng/mL MIP-1β for 12, 24, or 48 h. The apoptosis of the cells stimulated with MIP-1β (10, 20, and 40 ng/mL) for 24 h was analyzed using flow cytometry with Annexin V/PI double staining.
CCR5 expression was detected both on the membrane and in the cytoplasm in all the 3 tongue squamous cell carcinoma cell lines. At the concentrations of 10, 20, and 40 ng/mL, MIP-1β stimulation for 12 and 24 h significantly promoted the proliferation of CAL-27 cells (P<0.05); MIP-1β stimulation for 48 h at the concentrations 10 and 20 ng/mL, but not at 40 ng/mL, promoted the proliferation of CAL-27 cells (P<0.05). MIP-1β stimulation at 40 ng/mL for 24 produced the most obvious apoptosis-inducing effect in CAL -27 cells (P<0.05), while MIP-1β at 10 or 20 ng/mL did not induce obvious apoptosis in the cells (P>0.05).
CCR5 is expressed in all the 3 human tongue squamous cell carcinoma cells. MIP-1β can promote the proliferation of CAL-27 cells but high concentrations of MIP-1β also induced cell apoptosis. Prolonged stimulation of the cells with a high concentration of MIP-1β shows attenuated effect in promoting cell proliferation probably as a result of cell apoptosis induced by MIP-1β.
检测不同人舌鳞状细胞癌细胞中CCR5蛋白的表达,并观察巨噬细胞炎性蛋白-1β(MIP-1β)对CAL-27细胞增殖和凋亡的影响。
采用蛋白质免疫印迹法和免疫荧光染色法检测MIP-1β的受体CCR5在3种人舌鳞状细胞癌细胞UM-1、CAL-27和Tca-8113中的表达。采用CCK-8法评估用10、20和40 ng/mL MIP-1β刺激12、24或48 h后CAL-27细胞的增殖情况。使用Annexin V/PI双染流式细胞术分析用MIP-1β(10、20和40 ng/mL)刺激24 h后细胞的凋亡情况。
在所有3种舌鳞状细胞癌细胞系的细胞膜和细胞质中均检测到CCR5表达。在10、20和40 ng/mL浓度下,MIP-1β刺激12和24 h显著促进CAL-27细胞增殖(P<0.05);在10和20 ng/mL浓度下,MIP-1β刺激48 h促进CAL-27细胞增殖(P<0.05),但40 ng/mL浓度下未促进(P<0.05)。40 ng/mL MIP-1β刺激24 h对CAL-27细胞产生最明显的凋亡诱导作用(P<0.05),而10或20 ng/mL MIP-1β未诱导细胞明显凋亡(P>0.05)。
CCR5在所有3种人舌鳞状细胞癌细胞中均有表达。MIP-1β可促进CAL-27细胞增殖,但高浓度的MIP-1β也可诱导细胞凋亡。高浓度MIP-1β对细胞的长期刺激在促进细胞增殖方面显示出减弱的作用,这可能是由于MIP-1β诱导细胞凋亡所致。