Notch信号通路在小鼠肝癌细胞H22增殖凋亡中的作用及意义  

The role and significance of Notch signaling pathway in the proliferation and apoptosis of H22 cellsinmice

作　　者：陈勇[1] 金晓燕[1] 梁勇[1] 杨林军[1] 

机构地区：[1]台州学院医学院附属台州市立医院肿瘤外科,浙江台州318000

出　　处：《肝胆胰外科杂志》2012年第6期470-473,共4页Journal of Hepatopancreatobiliary Surgery

基　　金：2010年国家自然科学基金项目(81072209);2010年台州市科技项目(102ky11);2010年浙江省台州市椒江区科技项目(10278)

摘　　要：目的观察Notch信号通路在小鼠肝癌细胞H22增殖和凋亡中的作用,并探讨其可能的机制。方法培养小鼠肝癌细胞株H22,将细胞随机分为A、B、C组,分别加入Notch信号激活剂rhNF-κB、抑制剂DAPT及PBS共培养48 h。用MTT法检测细胞吸光度值(A值)观察细胞增殖情况,流式细胞术检测细胞凋亡率,应用免疫组化法检测各相关处理过的细胞株,RT-PCR、Western blotting法分别检测H22细胞中的Notch1、Hes1、Bcl-2 mRNA及其蛋白。结果培养24、48、72、96 h后,A、B组与C组比较,细胞增殖差异明显(P均<0.05)。A、B、C组细胞凋亡率分别为25.52%±2.13%、0.91%±0.18%、8.91%±1.50%,A、B组与C组比较差异明显(P均<0.05)。与C组比较,A组细胞中Notch1、Hes1、Bcl-2 mRNA及其蛋白表达量均显著增加(P均<0.05),B组细胞中Notch1、Hes1、Bcl-2 mRNA及其蛋白表达量均显著降低(P均<0.05)。结论 Notch1信号通路在抑制小鼠肝癌细胞株H22增殖、促进其凋亡中发挥了重要的调控作用,其机制可能与调节靶基因Hesl、Bcl-2表达有关。Objective To observe the role of Notch signaling pathway in the proliferation and apoptosis ofmouse hepatocellular carcinoma cell line H22, and to explore its possible mechanism. Methods Cultured H22 cells were randomly divided into A, B and C group, Notch signaling activator rhNF-KB, Notch signaling inhibitor DAPT, and PBS, were added separately into above three groups, then cultured together for 48 h. Cell proliferation was observed with MTT method （cell absorbance value）, cell apoptosis was detected with flow cvtometrv method, and the treated cells were analyzed with immunohistochemistrv method （fluorescence staining）. Notchl, Hesl, and Bcl-2 mRNA and the corre- sponding proteins in cultured cells were detected with RT-PCR and Western blotting methods, respectively. Results After 24, 48, 72 and 96 h of cell culture, the difference of cell proliferation among A, B, C group was significant （all P 〈0.05）. The apoptosis rates of A, B, C group were 25.52% ± 3.13%, 0.91% ± 0.18%, 8.91% ± 1.50%, respectively, the difference among three groups was significant （all P〈0.05）. Compared with group C, Notchl, Hesl, Bcl-2 mRNA and their protein expression in group A were siguificantlv increased （P〈0.05）, whereas the Notchl, Hesl, Bcl-2 mRNA and their protein expression in group B were siguificantlv decreased （P〈0.05）. Conclusion Notch signaling pathway plays an important role in the inhibition of cell proliferation and the promotion of cell apoptosis of mouse hepatocellular carcinoma cell line H22. The possible mechanism mac be related to its regulation of the expression of target gene Hesl and Bcl-2.

关 键 词：NOTCH信号通路 Hes1基因 BCL-2基因 细胞凋亡 细胞增殖 肝癌细胞H22 小鼠 

分 类 号：R735.7[医药卫生—肿瘤]