全反式维甲酸对食管癌EC109细胞骨架微丝的调节作用和细胞增殖的影响

The effects of all-trans retinoic acid on the cytoskeleton and growth of esophageal carcinoma cell line EC109

目的:观察全反式维甲酸(all-trans retinoic acid,atRA)对食管癌EC109细胞的增殖、凋亡以及细胞骨架微丝的调节作用。方法:不同浓度的atRA作用食管癌EC109细胞48 h后,相差显微镜观察细胞的生长状态,MTT检测细胞的增殖,Annexin V-FITC染色后通过流式细胞仪检测细胞的凋亡,荧光标记的鬼笔环肽染色观察细胞骨架的形态和分布。结果:0.1、1、2、5μmol/L atRA作用48 h后细胞形态均未见明显的异常。MTT检测显示,0.1、1、2、5μmol/L atRA作用组细胞的相对增殖比例(以对照组为1)分别为98.29%、93.15%、89.92%、81.03%,其中5μmol/L atRA作用组与对照组相比,差异具有统计学意义(P<0.05)。流式细胞仪检测0.1、1、2、5μmol/L atRA组早期凋亡细胞的比例分别为0.65%、0.94%、0.93%和0.94%,与对照组(0.70%)相比差异均无统计学意义(P>0.05)。荧光标记染色显示2μmol/L和5μmol/L atRA作用24 h后细胞骨架失去典型的丝状结构,出现排列紊乱。结论:atRA对食管癌EC109细胞无明显的凋亡诱导作用,但高浓度的atRA对食管癌细胞具有一定的生长抑制作用,并有可能通过细胞骨架调节细胞的分裂和增殖。

OBJECTIVE：To study the effects of all-trans retinoic acid（atRA） on the proliferation,apoptosis and cytoskeleton microfilaments of esophageal carcinoma cells EC109.METHODS： After exposed to atRA for 48 h,the morphological structure of EC109 cell were examined under phase contrast microscope,and MTT assay was applied to evaluate cell growth.The apoptic cells were stained with Annexin V-FITC and detected by flow cytometry.The cytoskeleton microfilaments were studied with fluorescence microscope after staining with FITC-labelled phalloidin.RESULTS： The proliferation ratios of 0.1,1,2,5 μmol/L atRA-treated group were 98.29%,93.15%,89.92% and 81.03%,respectively,with significant difference between 5 μmol/L atRA-treated group and control group（P0.05）.Cells in early stage of apoptosis of treated groups were 0.65%,0.94%,0.93%,0.94%,respectively,but without significant difference when compared with control group.The cytoskeleton was re-arranged and lost the typical filamentous structure after EC109 cells were exposed to 2 μmol/L and 5 μmol/L atRA for 24 h.CONCLUSION： Present study did not show atRA induced apoptosis of esophageal carcinoma cell line EC109,while atRA at high concentration demonstrated a potential to inhibit the growth of esophageal carcinoma cells.It is possible that atRA regulates the proliferation and growth of esophageal carcinoma cells through cytoskeleton changes.