DDX10和PTEN在肝癌大鼠进程中表达的变化

Changes of DDX10 and PTEN Expression in Hepatocarcinogenesis Rat

目的研究不同肝癌时期大鼠肝癌组织中DEAD-Box RNA解旋酶10(DDX10)和人第10号染色体缺失的磷酸酶—张力蛋白同源的基因(phosphatase and tensin homolog deleted on chromosome ten,PTEN)的变化和作用机制。方法用0.01%二乙基亚硝胺(diethylnitrosamine,DEN)溶液喂养SD大鼠诱发肝癌,建立大鼠肝癌模型,按大鼠喂养DEN的时间长短分为4组:正常组,肝癌早期组,肝癌中期组,肝癌晚期组。观察各组大鼠肝脏形态,通过血清学试剂盒法测定各组肝功能变化,采用苏木精-伊红(hematoxylin-eosin,HE)染色法观察各组大鼠肝癌组织的病理特征;并对肝癌组织进行免疫组织化学法染色,采用实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qPCR)检测DDX10和PTEN在肝脏组织中的表达情况;采用Western blot测定DDX10和PTEN的蛋白含量。结果肝癌中期组和肝癌晚期组大鼠肝功能与正常组大鼠比较均具有统计学意义(P<0.05)。HE染色结果表明各组大鼠的病理特征表现不同。免疫组织化学染色表明DDX10在肝癌晚期阳性表达率达到87.50%,明显高于其他组,而PTEN的阳性表达低于其他组,仅为37.50%;测得的DDX10mRNA和DDX10蛋白含量在肝癌晚期组中表达最高,而PTEN mRNA和PTEN蛋白含量在肝癌晚期组中表达最低,肝癌中期组和肝癌晚期组的DDX10蛋白含量、PTEN mRNA及PTEN蛋白含量与对照组比较,差异均有统计学意义(P<0.05)。结论在肝癌的发生发展中,DDX10可能起到致癌基因的作用,可以反映肝损伤的程度,而PTEN起到抑癌作用,它们与肝癌的相关性还有待进一步研究。

Objective To investigate the changes and mechanisms of DDX10 and phosphatase and tensin homolog deleted on chromosome ten(PTEN)in hepatocarcinogenesis rat liver tissue.Methods SD rats were fed with 0.01% diethylnitrosamine(DEN)solution to induce liver cancer,then rats were divided into normal group and early-stage,middlestage,late-stage liver cancer group according to the length of fed time.The morphology of the liver were observed,the changes of liver function in each group were determined by serological kit method.The pathological characteristics of liver cancer tissues in each group were observed by hematoxylin-eosin(HE)staining.The expression of DDX10 and PTEN in tissues was detected by quantitative real-time polymerase chain reaction(qPCR)after performing immunohistochemical staining of hepatocellular carcinoma tissues.The protein contents of DDX10 and PTEN were determined by Western blot.Results There was significantly different in the liver function between normal group and middle-stage or late-stage liver cancer group(P<0.05).HE staining showed that the pathological characteristics of each group were different.Immunohistochemistry showed that the positive expression rate of DDX10 in late-stage liver cancer group was 87.50%,which was obviously higher than that of the other groups,and the positive expression of PTEN was lower than that of other groups,only 37.50%.The expressions of DDX10 mRNA and DDX10 protein were highest,but the expression of PTEN mRNA and PTEN protein was lowest in late-stage liver cancer group when compared with other groups.The expressions of PTEN mRNA,DDX10 protein and PTEN protein in middle-stage and late-stage liver cancer groups were significantly different than those in normal group(P<0.05).Conclusion In the development of liver cancer,DDX10 may play a role in the oncogene,which can reflect the degree of liver damage,and PTEN plays a role in suppressing cancer.The correlation between them and liver cancer remains to be further studied.