摘要
目的:研究全反式维甲酸(all-trans-retinoicacid,ATRA)联合放射线照射对食管癌TE13细胞增殖、凋亡的影响及可能的作用机制。方法:采用MTT法检测ATRA对人食管癌细胞TE13增殖的影响;以其对TE13细胞生长抑制率(inhibitory rate,IR)达25%、50%和75%时的ATRA的浓度分别联合γ-射线进行分组处理细胞;FCM法分析ATRA单药及联合放射治疗对TE13细胞周期及凋亡的影响;采用克隆形成实验分析对克隆形成率和细胞存活率的影响;采用FCM法检测cyclinD1蛋白的表达。结果:ATRA对TE13细胞有增殖抑制作用,且呈剂量和时间依赖性;ATRA浓度为0.78、6.25和12.5μmol/L作用48h后,其对TE13细胞的IR分别达到22.0%、55.1%和71.1%。ATRA联合γ-射线(4Gy)与ATRA单药组比较,可明显降低TE13细胞的存活率和克隆形成率;ATRA联合γ-射线(4Gy)处理TE13细胞24和48h后,细胞增殖能力明显下降,更多的细胞被阻滞于G0/G1期,细胞凋亡率明显增加,与ATRA单药组比较,联合放射治疗可明显增强对TE13细胞周期和凋亡的影响;ATRA浓度为0.78μmol/L时和γ-射线联合作用对细胞周期cyclinD1蛋白表达量影响不大,但ATRA浓度为6.25和12.5μmol/L时则能明显下调cyclinD1的表达。结论:ATRA对TE13细胞具有明显的增殖抑制作用,其可能通过下调cyclinD1蛋白的表达,将TE13细胞阻断于G0/G1期并诱导细胞凋亡。在ATRA浓度较高时联合放疗,能显著下调cyclinD1蛋白表达,加强细胞G0/G1期阻滞及促进凋亡,抑制细胞克隆的形成能力;在ATRA浓度较低时联合放疗对原代细胞cyclinD1蛋白表达量的影响较小,但对促进细胞凋亡、抑制细胞克隆仍有一定的效果,其损伤效应可能需要一定时间。
Objective：To investigate the effects of ATRA （all-trans-retinoicacid） combined with gamma radiation on proliferation and apoptosis of human esophageal carcinoma TE13 cells, and to explore the possible mechanism. Methods：The effect of ATRA on the proliferation of TE13 cells was detected by MTT method. When the cell growth RI （inhibitory rate） reached levels of 25%, 50% and 75%, the TE13 cells were treated with the corresponding inhibitory doses of ATRA combined with 4 Gy gamma radiation. The effects of this combination intervention on cell cycle distribution and apoptosis of TE13 cells were detected by FCM （flow cytometry）. The colony-formation ability and cell viability were detected using colony-formation experiment. The expression of cyclinD1 protein was detected by FCM. Results：The inhibitory effect of ATRA on the proliferation of TE13 cells was significant in a dose- and time-dependent manner. The cell growth IRs reached 22.0%, 55.1% and 71.1% at ATRA concentrations of 0.78, 6.25 and 12.5 μmol/L, respectively. The cell viability and colony-formation efficiency were significantly decreased in TE13 cells treated with ATRA in combination with 4 Gy gamma radiation, as compared with TE13 cells receiving administration of ATRA alone. The proliferative ability of TE13 cells was significantly reduced after ATRA treatment in combination with 4 Gy gamma radiation for 24 and 48 h; furthermore, the percentage of the cells arrested at phase G0/G1 was increased accompanying with a significantly elevated apoptotic rate. Although the combination treatment （0.78 μmol/L ATRA and gamma radiation） had a weak influence on the expression of cyclinD1 protein, which was significantly decreased in other groups （6.25 and 12.5 μmol/L ATRA）. Conclusion：ATRA exerts an inhibitory influence on the proliferation of TE13 cells through down-regulating cyclinD1 expression, arresting the cells at phase G0/G1, and inducing apoptosis. A higher-concentration of ATRA combined with gamma radiation can significantly decrease the expression of cyclinD1, promote G0/G1 cell cycle arrest, accelerate apoptosis, and limit the colony formation, but a lower concentration of ATRA combined with gamma radiation exerts a little influence on cyclinD1 expression, although it may accelerate apoptosis and limit the colony-formation at some periods of time after treatment.
出处
《肿瘤》
CAS
CSCD
北大核心
2012年第8期592-598,共7页
Tumor
基金
河北省强势特色学科基金资助项目[编号:冀教高(2005)52号]
关键词
食管肿瘤
维甲酸
抗肿瘤联合化疗方案
放射疗法
Esophageal neoplasms
Tretinoin
Antineoplastic combined chemotherapy protocols
Radiotherapy
