摘要
目的探讨肝癌细胞和肝癌组织中同源盒基因(Homeobox genes)的表达特点及苯乙酸(PA)作用后肝癌细胞系SSMC-7721细胞HOX基因的表达变化。方法应用噻唑蓝(MTT)比色法,以0.25、0.5、1.0、2.0、4.0 mmol/L的PA作用于肝癌SSMC-7721细胞,分别于24、48、72 h对细胞增殖进行检测。设计不同的引物,将已知的22种HOX基因分为3组(P1、P2、P3),应用半定量逆转录-聚合酶链反应(RT-PCR)检测应用PA前后3组HOX基因mRNA的表达水平。HOX基因组表达水平用被检测基因组与β肌动蛋白(β-actin)灰度比值表示。结果PA对SSMC-7721的抑制作用呈现剂量-时间依赖性。二倍体细胞和正常肝组织中P1、P2、P3基因表达量相当,硬化肝组织和癌旁组织中P1表达增高(P<0.05),肝癌细胞HepG2、SSMC-7721和肝癌组织中P1显著增高(P<0.01),苯乙酸可抑制SSMC-7721细胞中P1基因的表达。结论肝癌组织和细胞中HOX基因组的P1组基因显著高表达,这种表达可被苯乙酸抑制。研究HOX基因的表达特点可能会对肝癌治疗的研究提供新的思路和方法。
Objective To study the characteristic expression of H0X gene in hepatocarcinoma tissue and cells and the change mediated by Phenylacetate. Methods SSMC-7721 cells grew in the presence or absence of PA （0.25,0.5,1.0,2.0,4.0 mmol/L） for 24 h, 48 h and 72 h, respectively. The cellular proliferation inhibitory ratio was evaluated by MTT assay. 22 kinds of Hox gene were divided into 3 groups according to their primer sequence. Semiquantitative reverse transeription-polymerase chain reaction（RT-PCR） used to investigate the mRNA expression of Hox gene groups and some kind of Hox gene in hepatoearcinoma tissue and cell lines following differentiation induced by PA. The level of Hox gene expression was expressed as ratio expression rate（RER） of Hox gene/β-aetin according to Computer Image analysis. We also detected the expression of P1 mRNA with the presentation of phenylaeetate. Results SSMC-7721 ceils were treated by 0.25 - 4.0 mmol/L PA for 24 - 72 h. With the increase of concentration of PA or the prolongation of the treating time, the proliferation inhibitory ratio of the tumor cells was increased notably to 91.9 % when treated for 72 h by 4. 0 mmoL/L. P1 was extremely high expressed in hepatocareinoma tissue and cell lines（ P 〈 0.01 ）. it also rised in hepatocirmsis and peri-tumor tissue（ P 〈 0.05）. Phenylacetate can inhabitate the expression of P1 mRNA. phenylaeetate. Conclusion P1 of HOX gene was extremely high expressed in hepatoearcinoma tissue and cell lines. P1 contributes the occurrence of liver tumor. PA could effectively inhibit the P1 expresion in SSMC-7721 cell.
出处
《中国实验诊断学》
北大核心
2009年第7期916-918,共3页
Chinese Journal of Laboratory Diagnosis
基金
吉林省科技发展项目资助课题(200505167)
关键词
同源盒基因
肝癌
表达特点
Homeobox genes
Hepatocarcinoma
characteristic expression
