α亚族趋化因子受体3在人舌鳞癌细胞株的表达及其配体干扰素诱导蛋白-10对CAL-27细胞增殖和凋亡的影响

目的探讨α亚族趋化因子受体3(CXC-chemokine receptor 3,CXCR3)在不同人舌鳞癌细胞株的表达以及其配体干扰素诱导蛋白-10(interferon induced protein 10,IP-10)对人舌鳞癌细胞株CAL-27增殖和凋亡的影响。方法采用蛋白印迹法和免疫荧光染色对3种人舌鳞癌细胞株(CAL-27、UM-1、Tca-8113)IP-10的相应受体CXCR3的表达进行检测。CAL-27细胞被分为3个实验组和1个对照组,3个实验组根据IP-10的刺激浓度,分为10 ng/mL组、20 ng/mL组、40 ng/mL组,对照组不予刺激。12 h、24 h、48 h时检测4组细胞的增殖;24 h时用流式细胞仪检测细胞的凋亡。结果 CXCR3在3种人舌鳞癌细胞株均有表达,并且CXCR3在3株细胞的胞膜及胞质内均有染色。和对照组相比,12 h、24 h时,3种浓度的IP-10均能够促进CAL-27细胞增殖(P<0.05);在48 h时,只有40 ng/mL的IP-10能够促进CAL-27细胞增殖(P<0.05),10 ng/mL、20 ng/mL的IP-10对CAL-27细胞的增殖无促进作用(P>0.05)。在24 h时,对照组、10 ng/mL组、20 ng/mL组和40 ng/mL组的细胞凋亡率分别为(0.053 3±0.472 6)%、(3.236 7±0.940 0)%、(4.516 7±1.115 4)%和(3.363 3±0.571 2)%,3种浓度IP-10均能够促进CAL-27细胞的凋亡(P<0.05),但3个实验组之间差异无统计学意义(P>0.05)。结论CXCR3在3种人舌鳞癌细胞株的胞膜及胞质内均有表达;IP-10既能促进CAL-27细胞的增殖,又能促进其凋亡。随着时间的延长,IP-10的促增殖作用减弱,而这种减弱可能是由IP-10的促凋亡作用的逐步发挥所引起的。

Chemokine receptor CXCR3 expression in different human tongue squamous cell carcinoma cell lines and the effect of IP-10/CXCL-10 on the proliferation and apoptosis of CAL-27 tongue cancer cells

To investigate the CXCR3 expression in different human tongue squamous cell carcinoma cell ]lines, and the effect of interferon-inducible protein-10 （P-I0） on the proliferation and apoptosis of CAL-27 cell. Meth- ods Respectively, Western blot and immunofluorescence staining were used to examin the expression of CXCR3 on 3 different human tongue squamous cell carcinoma cell lines （CAL-27, UM-1, Tca-8113）. Depending on the concentration of IP-10 stimulating, cells were divided into four groups： 0 ng/mL group （control group）, 10 ng/mL group, 20 ng/mL group, 40 ng/mL groups. Culturing CAL-27ceIIs in vitro with IP-10 （0 ng/＇mL, 10 ng/mL, 20 ng/mL, 40 ng/mL） , the proliferation with CCK-8 assay ＇after 12 h, 24 h, 48 h were observed. Annexin V/PI double staining flow cytometry detec- ted the apoptosis of CAL-27, ＇after 24 h treated with the same concentration gradient of IP-10. Results CXCR3 ex- pressed in three different human tongue squamous cell carcinoma ceils, stained in cell membrane and cytoplasm. Com- pared to the control group, 12 h and 24 h, different concentrations of IP-10 could promote the proliferation of CAL-27 （P〈0.05）; 48 h, only 40 ng/mL IP-10 could promote the proliferation of CAL-27 cell （P 〈0.05）, 10 ng/mL, 20 ng/mL IP-10 have no effect on the proliferation. After 24 h, apoptosis rate of control group, 10 ng/mL group, 20 ng/mL group and 40 ng/mL group were （0.053 3 ± 0.472 6） %, （ 3. 236 7± 0.940 0） %, （4.516 7 ± 1.115 4） % and （ 3. 363 3± 0.571 2 ） % respectively. IP-I 0 had a pm-apoptotic role on CAL-27 cells （ P 〈 0.05 ）, but there was no significant difference between the different concentration groups （P 〉 0.05）. Cortelusion IP-10 not only could promote CAL-27 cell proliferation, but also could promote apoptosis. With the extension of the duration of action, CAL-27 cell proliferation rate declines. The decline may be caused by gradually assuming IP-10 pro-apoptotic role.