Fang Jianlin, Zheng Chuansheng, Tao Hongfang, Zhao Hui, Ren Jianzhuang, Feng Gansheng
Department of Radiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
J Huazhong Univ Sci Technolog Med Sci. 2010 Feb;30(1):113-8. doi: 10.1007/s11596-010-0121-5. Epub 2010 Feb 14.
In order to investigate the inhibitory effects of all trans-retinoic acid (ATRA) on differentiation and apoptosis of Walker-256 hepatocellular carcinoma cells and the therapeutic effects of ATRA combined with transarterial chemoembolization (TACE) on rat Walker-256 transplanted hepatocarcinoma, Walker-256 hepatocarcinoma cell lines were treated with ATRA at different concentrations. After culture for 48 h, the inhibitory rate of cell proliferation was determined by MTT assay; the changes of Fas and Bcl-2 mRNA expression were determined by RT-PCR, and the expression levels of Caspase3 and Caspase8 proteins were detected by Western blot. Twenty-seven Wistar rat models of hepatocarcinoma were set up successfully by implanting Walker-256 cell lines. The tumor volume at the 11th day after implantation (V(preoperation)) was measured by magnetic resonance imaging (MRI). The 27 rats were randomly and equally divided into three groups, and the therapy scheme was performed as follows: group A (ATRA 0.1 mg+mitomycin 0.05 mL+lipiodol 0.05 mL+gelfoam powder 0.025 mg); group B (mitomycin 0.05 mg+lipiodol 0.05 ml+gelfoam 0.025 mg; group C (0.9% NaCl 0.2 mL). After another 11 days, MRI was performed once again to measure the tumor volume (V(postoperation)). The expression of factor and Ki VIII -67 in the tumor tissues was detected by immunohistochemistry. The results showed that ATRA could suppress proliferation of Walker-256 cell lines. After treatment of Walker-256 cell lines with ATRA, the expression of Fas mRNA was significantly up-regulated and the Bcl-2 mRNA was significantly down-regulated by ATRA at the concentration of 10 mumol/L as compared with the control group (P<0.05). After treatment with 10 mumol/L ATRA for 48 h, the Caspase3 and Caspase8 were significantly activated as compared with the control group (P<0.05). Significant difference existed in growth rate among the three groups (P<0.01) and between either two groups (P<0.05). The expression rate of factor VIII and Ki-67 was gradually increased from group A, group B to group C. The study suggests that ATRA could inhibit the proliferation of Walker-256 cells and the effectiveness of the combined therapy (ATRA+TACE) for treating transplanted hepatoma of rats is superior to that of TACE alone.
为探讨全反式维甲酸(ATRA)对Walker-256肝癌细胞分化及凋亡的抑制作用,以及ATRA联合肝动脉化疗栓塞术(TACE)对大鼠Walker-256移植性肝癌的治疗效果,用不同浓度的ATRA处理Walker-256肝癌细胞系。培养48 h后,采用MTT法测定细胞增殖抑制率;采用RT-PCR法检测Fas和Bcl-2 mRNA表达变化,采用Western blot法检测Caspase3和Caspase8蛋白表达水平。通过植入Walker-256细胞系成功建立27只Wistar大鼠肝癌模型。植入后第11天用磁共振成像(MRI)测量肿瘤体积(V(术前))。将27只大鼠随机均分为3组,治疗方案如下:A组(ATRA 0.1 mg+丝裂霉素0.05 mL+碘油0.05 mL+明胶海绵粉0.025 mg);B组(丝裂霉素0.05 mg+碘油0.05 ml+明胶海绵0.025 mg);C组(0.9%氯化钠0.2 mL)。再过11天后,再次行MRI测量肿瘤体积(V(术后))。采用免疫组织化学法检测肿瘤组织中因子和Ki VIII -67的表达。结果显示,ATRA可抑制Walker-256细胞系增殖。与对照组比较,用10 μmol/L ATRA处理Walker-256细胞系后,10 μmol/L ATRA组Fas mRNA表达显著上调,Bcl-2 mRNA显著下调(P<0.05)。用10 μmol/L ATRA处理48 h后,与对照组比较,Caspase3和Caspase8显著激活(P<0.05)。三组间生长率差异有统计学意义(P<0.01),任意两组间差异有统计学意义(P<0.05)。因子VIII和Ki-67表达率从A组、B组到C组逐渐升高。研究表明,ATRA可抑制Walker-256细胞增殖,ATRA联合TACE治疗大鼠移植性肝癌的疗效优于单纯TACE。