CAJ | 학술논문

目的观察鼻咽癌细胞潜伏膜蛋白1(LMP1)表达对β-catenin的转录活性及蛋白表达的影响,探讨LMP1与Wnt/β-catenin信号通路在鼻咽癌发生发展中的作用.方法应用免疫组织化学EnVision法检测75例鼻咽癌组织中LMP1和β-catenin的表达.构建pHA2-LMP1重组质粒,应用细胞免疫荧光、双荧光素酶报告分析、Western blot方法研究LMP1在鼻咽癌细胞株CNE1和CNE2中对β-catenin转录活性及表达的影响.结果 (1)鼻咽癌组织中β-catenin的异常表达率为50.7%(38/75),LMP1的阳性表达率为50.7%(38/75).鼻咽癌组织中β-catenin异常表达与LMP1表达存在正相关(相关系数为x2=7.048,P=0.008).(2)LMP1表达明显提高鼻咽癌细胞株CNE1和CNE2中β-catenin的核表达,且β-catenin的核表达在低分化鼻咽癌细胞株CNE2明显高于高分化鼻咽癌细胞株CNE1.(3)LMP1可明显上调CNE1和CNE2中β-catenin的转录活性,且呈时间依赖性.另外,β-catenin的转录活性在低分化鼻咽癌细胞株CNE2高于高分化鼻咽癌细胞株CNE1.(4)LMP1对鼻咽癌细胞株β3-catenin的总蛋白表达水平无明显影响.结论 EB病毒编码的LMP1可能通过β-catenin信号通路参与鼻咽癌的发生发展.
목적 관찰비인암세포잠복막단백1(LMP1)표체대β-catenin적전록활성급단백표체적영향,탐토LMP1여Wnt/β-catenin신호통로재비인암발생발전중적작용.방법응용면역조직화학EnVision법검측75례비인암조직중LMP1화β-catenin적표체.구건pHA2-LMP1중조질립,응용세포면역형광、쌍형광소매보고분석、Western blot방법연구LMP1재비인암세포주CNE1화CNE2중대β-catenin전록활성급표체적영향.결과 (1)비인암조직중β-catenin적이상표체솔위50.7%(38/75),LMP1적양성표체솔위50.7%(38/75).비인암조직중β-catenin이상표체여LMP1표체존재정상관(상관계수위x2=7.048,P=0.008).(2)LMP1표체명현제고비인암세포주CNE1화CNE2중β-catenin적핵표체,차β-catenin적핵표체재저분화비인암세포주CNE2명현고우고분화비인암세포주CNE1.(3)LMP1가명현상조CNE1화CNE2중β-catenin적전록활성,차정시간의뢰성.령외,β-catenin적전록활성재저분화비인암세포주CNE2고우고분화비인암세포주CNE1.(4)LMP1대비인암세포주β3-catenin적총단백표체수평무명현영향.결론 EB병독편마적LMP1가능통과β-catenin신호통로삼여비인암적발생발전.
Objective To investigate the contribution of latent membrane protein (LMP) 1 to nasopharyngeal carcinogenesis via Wnt/β-catenin signal pathway. Methods The recombinant plasmid pHA2-LMP1 was constructed; immunofluorescence staining, Dual-Luciferase Reporter Assay, Western blot and immunohistochemistry staining were used to study the effect of LMP1 on the transcriptional activity and expression of β-catenin. Results (1) Abnormal expression of β-catenin was obtained in 38 cases (50. 7% ,38/75), LMP1 expression was obtained in 38 cases (50. 7%, 38/75). There was significantly positive correlation between LMP1 expression and abnormal expression of β-catenin in nasopharyngeal carcinoma tissue (P =0.008). (2) The expression of β-catenin in nuclei of NPC cell line CNE1 and CNE2transfected with pHA2-LMP1 plasmid dramatically increased, and the expression was remarkable in poorlydifferentiated NPC cell line CNE2 than that of well-differentiated CNE1 cells. (3) LMP1 expression dramatically increased the transcriptional activity of β-catenin in CNE1 and CNE2 cells transfected with pHA2-LMP1 and was in a time-dependent. The transcriptional activity of β-catenin was higher in poorlydefferentiated cell line CNE2 than that of well-differentiated NPC cell line CNE1. (4) LMP1 expression did not affect the total protein expression level of β-catenin in both CNE1 and CNE2 cell lines. Conclusion EB virus-encoded LMP1 may be involved in the pathogenesis of NPC via β-catenin signal pathway.