免疫荧光法检测全反式维甲酸和As2O3对人口腔鳞状细胞癌KB细胞的诱导分化作用

Immunofluorescence examination of CK-13 expression in cell line KB differentiated by all-trans retinoic acid or As2O3

目的研究细胞角蛋白（cytokeratin，CK）13在口腔鳞状细胞癌中表达的变化规律，探讨全反式维甲酸（a11-transretinoic acid，ATRA）与三氧化二砷（As2O3）在诱导分化人口腔未分化鳞状细胞癌细胞系KB细胞中的作用。方法采用免疫荧光标记CK-13单克隆抗体及KB细胞细胞核，检测CK-13在ATRA与As2O3诱导分化KB细胞前后的表达变化。将同期培养的KB细胞分为对照组（细胞培养液）、ATRA实验组及As2O3实验组。培养72h后分别观察3个视野下对照组与实验组KB细胞中CK-13在细胞质中的表达情况，并统计胞质着色率。结果免疫荧光检测法发现对照组的KB细胞中CK-13绿色荧光染色胞质着色率为（1．00±0．58）％；ATRA实验组的3个浓度组中CK-13绿色荧光染色胞质着色率分别为（100．00±0．00）％、（100．00±0．00）％、（98．96±1．04）％，而As2O3，实验组的3个浓度组结果分别为（100．00±0．00）％、（100．00±0．00）％、（88．64±11．36）％。对照组与实验组CK-13绿色荧光染色胞质着色率的统计数据对比差异均有统计学意义（P〈0．05），各实验组之间相比差异无统计学意义（P〉0．05）。结论ATRA与As2O3对KB细胞均存在诱导分化作用，CK-13定位着色于细胞质，其表达变化与肿瘤细胞分化程度呈正相关，可作为评估ATRA和As2O3对OSCC诱导分化疗效的分子标志物。

Objective To examine the expression of cytokeratin-13 （CK-13） in oral squamous cell carcinoma （OSCC） and to discuss the effects of all-trans retinoic acid （ATRA） or arsenic trioxide （ As2O3 ） on the differentiation of human oral undifferentiated squamous cell carcinoma cell line KB cells. Methods The cultured KB cells were divided into three groups, ATRA group, As2O3 group, and control. The expression of CK-13 in KB cells was detected using the immunofluoreseence before and after KB cells were induced by ATRA or As2O3. Results The expression rates of CK-13 in KB cells in the ATRA group and As2O3 group were significantly higher than that in the control （ P 〈 0.05 ）, but there was no significant difference in the expression between ATRA and As2O3 group（P 〉0. 05）. Conclusions ATRA and As2O3 both have the ability to differentiate the KB cells, and the expression is associated with the degree of tumor differentiation. CK-13 may serve as a molecular marker to evaluate the effect of the differentiation treatment on OSCC.