Li Jie, Zhou Tao, Liu Liang, Ju Ying Chao, Chen Yue Tong, Tan Zi Rui, Wang Jing
Division of Medical Affairs, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, PR China.
Breast Disease Center, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, PR China.
Pathol Res Pract. 2018 Oct;214(10):1719-1725. doi: 10.1016/j.prp.2018.09.009. Epub 2018 Sep 13.
The following study investigated the effects of Annexin A3 (ANXA3) on breast cancer biological behavior in vivo, using nude mouse model bearing a subcutaneous tumor. A total of 18 female nude mice were randomly divided into three groups (n = 6): negative control group which was inoculated with MDA-MB-231 cells, blank control group which was inoculated with MDA-MB-231-NC cells, and the transfection group which was inoculated with MDA-MB-231-Sh cells. The experiment lasted for 4 weeks, during which mice conditions, diet and defecation were monitored on a daily basis. Body weight, as well as tumor diameters, which were assessed using standard caliper method, were measured once a week. In vivo imaging was performed to detect the activity of transplanted tumors. H&E staining was used to analyze the histological structure of tumor tissues in three groups, while flow cytometry and fluorescent RT-PCR were performed to measure cell proliferation and the expression of ANXA3 mRNA. Briefly, significantly slower tumor growth and tumor activity were observed in the transfection group compared to negative and blank controls, while the tumor weight and volume in this group were also significantly lower compared to the other two groups (P < 0.01). Sparse tumor cells accompanied with massive fibrous connective tissue proliferation, and lower new blood vessels formation were observed in transfection group compared to other groups. Moreover, mRNA and protein levels of ANXA3 were significantly lower in transfection group compared to the other two groups (P < 0.01). In addition, lower proliferation index and higher G cell count were observed in transfection group compared to negative and blank controls (P < 0.01). To sum up, these results suggested that ANXA3 silencing regulates the proliferation and inhibits the growth of MDA-MB-231 breast cancer cells. Consequently, ANXA3 might be used as a potential target for gene therapy in breast cancer.
以下研究利用携带皮下肿瘤的裸鼠模型,在体内研究了膜联蛋白A3(ANXA3)对乳腺癌生物学行为的影响。总共18只雌性裸鼠被随机分为三组(n = 6):接种MDA-MB-231细胞的阴性对照组、接种MDA-MB-231-NC细胞的空白对照组以及接种MDA-MB-231-Sh细胞的转染组。实验持续4周,在此期间每天监测小鼠的状况、饮食和排便情况。每周测量一次体重以及使用标准卡尺法评估的肿瘤直径。进行体内成像以检测移植肿瘤的活性。采用苏木精-伊红(H&E)染色分析三组肿瘤组织的组织结构,同时进行流式细胞术和荧光逆转录聚合酶链反应(RT-PCR)以测量细胞增殖和ANXA3 mRNA的表达。简而言之,与阴性对照组和空白对照组相比,转染组的肿瘤生长和肿瘤活性明显较慢,并且该组的肿瘤重量和体积也明显低于其他两组(P < 0.01)。与其他组相比,转染组观察到肿瘤细胞稀疏,伴有大量纤维结缔组织增生,且新血管形成较少。此外,与其他两组相比,转染组中ANXA3的mRNA和蛋白质水平明显较低(P < 0.01)。另外,与阴性对照组和空白对照组相比,转染组的增殖指数较低,G期细胞计数较高(P < 0.01)。综上所述,这些结果表明沉默ANXA3可调节MDA-MB-231乳腺癌细胞的增殖并抑制其生长。因此,ANXA3可能作为乳腺癌基因治疗的潜在靶点。