CAJ | 학술논문

目的观察NOTCH1/HES1/PTEN信号通路在浸润性膀胱癌组织中的表达情况,以及膀胱癌细胞中过表达NOTCH1对下游基因HES1与PTEN的表达及生物学的影响.方法运用荧光定量PCR和Western印迹技术,检测了36例浸润性膀胱癌组织和10例正常膀胱黏膜组织中NOTCH1、HES1和PTEN的表达情况.在膀胱癌细胞T24中转染NOTCH1表达质粒,检测各组细胞的NOTCH1、HES1、PTEN在mRNA和蛋白质水平的表达改变.采用MTS法和流式细胞技术检测各组细胞的增殖、凋亡与周期情况的变化.结果较正常膀胱黏膜组织,浸润性膀胱癌组织中NOTCH1表达水平为正常膀胱黏膜组织的4.22倍,IIES1为正常组的3.75倍.而浸润性膀胱癌组织中PTEN的表达显著下调,为正常组的38.96％(P＜0.05).过表达NOTCH1的T24细胞较对照组绌胞,HES1表达上调而PTEN下调(P＜0.01).MTS实验显示,转染NOTCH1表达质粒的细胞增殖能力较对照组细胞升高(P＜0 01).结论 NOTCH1的异常高表达是浸润性膀胱癌的分子事件之一,提示NOTCH1在浸润性膀胱癌中可能发挥癌基因的作用.
목적 관찰NOTCH1/HES1/PTEN신호통로재침윤성방광암조직중적표체정황,이급방광암세포중과표체NOTCH1대하유기인HES1여PTEN적표체급생물학적영향.방법 운용형광정량PCR화Western인적기술,검측료36례침윤성방광암조직화10례정상방광점막조직중NOTCH1、HES1화PTEN적표체정황.재방광암세포T24중전염NOTCH1표체질립,검측각조세포적NOTCH1、HES1、PTEN재mRNA화단백질수평적표체개변.채용MTS법화류식세포기술검측각조세포적증식、조망여주기정황적변화.결과 교정상방광점막조직,침윤성방광암조직중NOTCH1표체수평위정상방광점막조직적4.22배,IIES1위정상조적3.75배.이침윤성방광암조직중PTEN적표체현저하조,위정상조적38.96％(P＜0.05).과표체NOTCH1적T24세포교대조조출포,HES1표체상조이PTEN하조(P＜0.01).MTS실험현시,전염NOTCH1표체질립적세포증식능력교대조조세포승고(P＜0 01).결론 NOTCH1적이상고표체시침윤성방광암적분자사건지일,제시NOTCH1재침윤성방광암중가능발휘암기인적작용.
Objective To explore the role of NOTCH1/HES1/PTEN signaling pathway in invasive TCCB (bladder transitional cell carcinoma).Methods The expressions of NOTCH1,HES1 and PTEN were detected in 36 cases of invasive TCCB tissues and 10 cases of normal bladder samples by real-time q-polymerase chain reaction (q-PCR) and Western blot. Then NOTCH1-ORF plasmid and its blank vector pCMV6-Entry were transfected into T24 cell respectively. And the expressions of three above-mentioned target genes were measured by real-time q-PCR and Western blot. Furthermore,cell proliferation,cell apoptosis and cell cycle were analyzed respectively by MTS assay and flow cytometry.Results Compared with normal bladder samples,the higher levels of both mRNA and protein of NOTCH1 and HES1 were detected in invasive TCCB tissues while there was a lower expression of PTEN (P＜0.05).The mRNA expression levels of NOTCH1 and HES1 were 4.22 and 3.75 folds respectively higher than those of normal tissues.In NOTCH1-overexpressed T24 cell,the expression of HES1 was 5.43 folds higher than that of the blank vector control group while the expression of PTEN declined to 41.76％ (P＜0.01).MTS assay showed that the NOTCH1-ORF transfection obviously promoted cell proliferation in T24 cell (P＜0.01).Conclusion NOTCH1 gene may function as an oncogene by regulating HES1/PTEN in invasive TCCB and its aberrant activation promotes cell proliferation.