摘要:
目的 探讨PTEN基因表达变化在三羟异黄酮(genistein)抑制裸鼠移植入肝癌增长中的作用及其机制.方法 将移植入肝癌裸鼠分为2组,对照组腹腔注入含0.04%二甲基亚砜(DMSO)的RPMI 1640培养基每天0.05 ml/g,Genistein组腹腔注入genistein每天1 mg/kg,3周后观察肝癌增长,并应用同位素试剂盒检测肝癌组织三磷酸肌醇( IP3)含量,逆转录-聚合酶链反应(RT-PCR)分析癌组织PTEN mRNA表达,Western blot分析肝癌组织PTEN蛋白表达.结果 Genistein组肝癌体积和重量均显著低于对照组,其中体积为(12.6±11.6) mm3比(52.3±26.5) mm3,重量为(42.7±27.8) mg比(91.3±31.4) mg,IP3含量显著低于对照组,为(13.4±1.4) pmol/mg蛋白比(35.3±6.6)pmol/mg蛋白,PTEN mRNA表达显著高于对照组,灰度与面积之积的相对强度(RI)为0.81±0.24比0.36±0.09,PTEN蛋白表达显著高于对照组,RI值为3.14±0.13比1.08±0.15.结论 PTEN基因表达上调在Genistein抑制裸鼠移植人肝癌增长中发生一定作用,其机制可能与抑制磷酸肌醇通路信号转导、抑制IP3生成有关.%Objective To probe into the Role and mechanism of phosphatase and tensin homology deleted on chromosome ten (PTEN) gene expression change in inhabiting hepatocellular carcinoma of nude mice by genistein.Methods Animals with hepatocellular carcinoma were treated with genistein 1 mg/kgeveryday(ip) for 3 weeks,the volume and weight of tumaor was measured,Trisphosphate Inositol ( IP3 ),PTEN mRNA,PTEN protein were assayed by IP3-[ 3H] Birtrak assay,reverse transcription-polymerase chain reaction (RT-PCR),Western blotting,respectively.Results The tumor volume and weight of animals treated with genistein was lower than control [ Volume:( 12.6 ± 11.6) mm3 vs.(52.3 ± 26.5 )mm3,Weight:(42.7 ±27.8) mg vs.(91.3 ±31.4) mg],IP3 continent was lower than that of control [ ( 13.4± 1.4) pmol/mg protein vs.( 35.3 ± 6.6) pmol/mg protein),PTEN mRNA expression was higher in group treated with genistein than that of control,the RI which was the gray degree multiply area of PTEN/the gray degree multiply area of β-actin was 0.81 ± 0.24 vs.0.36 ± 0.09,PTEN mRNA expression was higher in group treated with genistein than that of control(3.14 ±0.13 vs.1.08±0.15).Condusion Genistein can inhabit growth of hepatocellular carcinoma tansplanted into liver of nude mice by up-regulating PTEN gene expression which is relative to signal transduction of phosphate Inositol leading to reduce IP3 production.