全反式维甲酸诱导喉癌细胞凋亡的实验研究

Study of all-trans retinoic acid-induced apoptosis of Hep-2 laryngeal carcinoma cells

目的探讨全反式维甲酸(All-trans retinoic acid,ATRA)诱导人喉鳞状细胞癌细胞株Hep-2产生凋亡及对角蛋白基因13(Cytokeratin 13,CK13)mRNA表达的影响。方法ATRA作用Hep-2细胞72 h后,分别用光学显微镜、电镜观察细胞形态及超微结构改变;并用逆转录聚合酶链反应(RT-PCR)方法检测CK13mRNA的表达。结果ATRA作用72 h后的Hep-2细胞在光学显微镜及电镜下细胞逐渐变圆,生长减缓、细胞表面绒毛消失,核染色质致密,浓缩成块,胞质中有空泡形成,细胞表面出泡,脱落形成膜包裹的凋亡小体等细胞凋亡典型的形态学改变;CK13 mRNA表达水平随ATRA浓度的增高而逐渐上调(P<0.05)。结论ATRA可以诱导喉癌细胞凋亡及CK13mRNA表达上调,并呈时间和浓度依赖性。

Objective To observe the influence of all-trans retinoic acid（ATRA）on the growth of Hep-2 laryngeal carcinoma cell line and cytokeratin13 （CK13） gene expression. Methods Hep-2 cell line was treated with ATRA for 72 h. Changes of the cell morphology and the ultra-structure were observed with optical microscope and electron microscope. Reverse transcription-polymerase chain reaction （RT-PCR） and immunocytochemical technique were used for detecting the expression of CK13 gene. Results After treated with ATRA for 72 hours, the Hep-2 cells grew slowly and the morphological examination with electron microscope showed the presence of cells with chromatin condensation and agglomerate, nuclear fragment, cell shrinkage, and ＂apoptotic bodies＂. The expression of CK13 gene mRNA could be up-regulated gradually with the increase of concentration of ATRA. The results were statistically significant （ P〈0. 05）. Conclusion All-trans retinoic acid can induce not only apoptosis of laryngeal carcinoma cells but also up-graduation of CK13 mRNA expression. The process depends on both timing and concentration of ATRA.