肌醇六磷酸诱导人结肠癌细胞系HT-29细胞分化的实验研究  

THE EXPERIMENTAL STUDY OF CELL-INDUCED DIFFERENTIATION BY INOSITOL HEXAPHOSPHATE IN HT-29 HUMAN COLON CARCINOMA CELL LINE

作　　者：刘晓燕[1] 宋扬[1] 张晓妮[1] 

机构地区：[1]青岛大学医学院营养与食品卫生研究所,青岛266021

出　　处：《营养学报》2009年第4期388-391,共4页Acta Nutrimenta Sinica

基　　金：国家自然科学基金(No.30671767);山东省自然科学基金项目(No.2007BS03042)

摘　　要：目的使用不同浓度的肌醇六磷酸(IP6)作用于人结肠癌细胞系HT-29,观察IP6对癌细胞分化的影响,并对其机制进行探讨。方法将不同浓度的IP6(对照,1.8mmol/L,3.3mmol/L)作用HT-29细胞不同时间(1d,3d,5d),分别进行如下研究:(1)使用透射电镜观察细胞超微结构的改变;(2)测定碱性磷酸酶比活性;(3)应用RT-PCR法检测细胞内c-Myc基因的表达;(4)使用免疫细胞化学法检测细胞内Rb蛋白的表达。结果(1)3.3mmol/LIP6持续作用后,透射电镜下可以观察到细胞结构发生改变,出现核固缩、细胞器丰富、微绒毛增多等分化趋势;(2)IP6作用HT-29细胞后,碱性磷酸酶比活性明显增强;(3)RT-PCR结果显示,与对照组相比,各浓度IP6均能使细胞内c-Myc mRNA的表达减少(P<0.05);(4)IP6能够上调Rb蛋白的表达(P<0.05)。结论IP6能够促使HT-29细胞向正常细胞转化。IP6可能通过上调Rb蛋白的表达,下调c-Myc蛋白的表达,阻滞细胞周期,诱导细胞分化而发挥其抗肿瘤作用。Objective To investigate the effects and mechanisms of inositol hexaphosphate on differentiation of HT-29 human colon carcinoma cell line. Method Cells were exposed to various concentrations （control, 1.8 mmol/L, 3.3 mmol/L） of IP6 for different time （ld, 3d, 5d）. Its effects on the cells were evaluated by detecting follow indices： （1） Observing ultrastructure of HT-29 ceils by transmission electron microscopy. （2） Detecting alkaline phosphate （ALP） activity to evaluate cell differentiation. （3） Expression of c-Myc mRNA was determined by reverse transcription polymerase chain reaction （RT-PCR）. （4） Immunocytochemical stain was used to detect expression of Rb protein. Results （1） After having been treated at 3.3mmol/L concentration, the cell structure changed, and appeared tendency of differentiation, such as karyopyknosis, abundant cellular organs and microvillus increase. （2） The activity of alkaline phosphate increased along with the concentration and time. （3） RT-PCR showed that different concentrations of IP6 decreased the expression of c-Myc mRNA. （4） The immunocytochemical stain showed that different concentrations of IP6 increased expression of Rb protein. Conclusion IP6 Can induce differentiation of HT-29 cell by augmenting expression of Rb, decreasing expression of c-Myc and blocking cell cycle.

关 键 词：肌醇六磷酸 HT-29细胞系 细胞分化 

分 类 号：R735.35[医药卫生—肿瘤]