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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 investigate the effects of all-trans retinoic acid (ATRA) on the apoptosis of human laryngeal squamous cell carcinoma cell line Hep-2 and on the expression of cytokeratin 13 (CK13) mRNA. Methods After treated with ATRA for 72 h, the morphology and ultrastructure of Hep-2 cells were observed by light microscopy and electron microscopy. The expression of CK13 mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Results Hep-2 cells after ATRA treatment for 72 h gradually became round under the light microscope and electron microscope. The growth of the Hep-2 cells was slowed down. The surface villi disappeared. The chromatin of the nucleus became dense and concentrated. The vacuoles formed in the cytoplasm. (P <0.05). The typical morphological changes of apoptotic bodies such as apoptotic bodies were observed by flow cytometry. The level of CK13 mRNA was up-regulated with the increase of ATRA concentration (P <0.05). Conclusion ATRA can induce the apoptosis of laryngeal carcinoma cells and up-regulate the expression of CK13 mRNA in a time and concentration-dependent manner.