目的:通过体外实验探讨Wnt2B在乳腺癌发生发展中的作用.方法:应用免疫组织化学的方法检测正常乳腺组织和乳腺癌组织中Wnt2B的表达；采用RNA干扰技术,在MCF7细胞中沉默Wnt2B的表达,并经Western blot检测其沉默效率；应用流式细胞术和激光共聚焦显微镜检测、观察Wnt2B沉默后对MCF7细胞凋亡、细胞骨架构成的改变.同时在蛋白水平上检测沉默Wnt2B后对细胞P53/P21通路的影响.结果:Wnt2B蛋白在乳腺癌中的表达明显高于在正常乳腺组织中的表达；第2对Wnt2B siRNA(2号siRNA)对Wnt2B的沉默效果最为明显；沉默Wnt2B后细胞早期凋亡率和晚期凋亡率相对于对照细胞均明显增高并伴有细胞骨架明显的改变,表现为细胞骨架溶解,正常形态消失.同时,细胞内P53及P21表达水平随Wnt2B沉默而明显下降.结论:Wnt2B基因可以改变乳腺癌细胞的凋亡水平和骨架结构,在乳腺癌的发生发展中起着重要的作用,为乳腺癌的治疗提供了新的靶点.%Objective To explore the function of Wnt2B in the occurrence and development of breast cancer in vitro. Methods The expression of Wnt2B in normal breast tissue and breast cancer tissue were detected by immnohistochemistry. The expression of Wnt2B was silenced in MCF7 cells by RNA interference and the rate of silencing Wnt2B was detected by Western blot. Changes of apoptosis and cytoskeleton of MCF7 after silencing Wnt2B was detected and observed by flow cytometry and laser scanning confocal microscope and effects on the pathway of P53/P21 after silencing Wnt2B was measured in protein level. Results Immunohistochemistry results showed that Wnt2B had significantly higher expression rate in breast cancer tissue than that in normal breast tissue. The second pair of siRNA had the strongest capability to silence Wnt2B. Apoptosis rate in the first period and in the later period after silencing Wnt2B were increased significantly compared with control group and changes of skeleton structure of cells were obvious like dissolution of cytoskeleton and disappearance of normal structure. Meanwhile, with the silencing Wnt2B, the expression of P53 and P21 were reduced significantly. Conclusions Wnt2B can regular apoptosis and skeleton structure of breast cancer cells. It plays an important role in the occurrence and development of breast cancer and provides new targets for the treatment of breast cancer.
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