Chinese Medical E-ournals Database

Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition) ›› 2016, Vol. 12 ›› Issue (03): 274 -279. doi: 10.3877/cma.j.issn.1673-5250.2016.03.005

Special Issue:

Original Article

Influence of annexin A2 expression regulated by E6-associated protein on trip-negative breast cancer in nude mouse with transplantation tumor

Lingmi Hou1, Shaoli Xie2, Maoshan Chen3, Jingdong Li4, Ajedichiga Aduah Emmanuel4, Hongwei Yang3, Yanchun Gao2, Tianyong Xing2, Shishan Deng5, Xiaobo Zhao2,()   

  1. 1. Department of Thyroid and Breast Surgery; Institute of Hepatobiliary and Pancreatic-intestinal Diseases, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China
    2. Department of Thyroid and Breast Surgery
    3. Department of Breast and Thyroid Surgery, Suining Central Hospital, Suining 629000, Sichuan Province, China
    4. Institute of Hepatobiliary and Pancreatic-intestinal Diseases, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China
    5. Department of Anatomy, North Sichuan Medical College, Nanchong 637000, Sichuan Province, China
  • Received:2015-11-01 Revised:2016-05-10 Published:2016-06-01
  • Corresponding author: Xiaobo Zhao
  • About author:
    Corresponding author: Zhao Xiaobo, Email:
Objective

To study the influence of annexin A2 expression regulated by E6-associated protein (E6-AP) on trip-negative breast cancer (TNBC) in nude mouse with transplantation tumor.

Methods

From April to November 2014, 40 female BALB/C-nu nude mice were selected as research subjects. They were divided into experiment group (n=20) and control group (n=20) according to random number table method. The construction of TNBC nude mice with transplantation tumor models in two groups: E6-AP-RNA interference target sequences were designed, and then small interfering RNA (siRNA) fragments were transfected into TNBC cell lines MDA-MB-231, and these TNBC cell lines were injected subcutaneously into nude mice in experimental group. While TNBC cell lines MDA-MB-231 which were not transfected with E6-AP siRNA were injected subcutaneously into control group. The transplanted tumor formations in two groups were observed. Relative expressions of E6-AP and annexin A2 in two groups were assayed through Western blotting method. And positive expression rate of annexin A2 in transplanted tumor of two groups were assayed by immunohistochemical method. And those indexes above were analyzed by statistical methods. The disposal of animals in this research was in accordance with the animal ethical standards. There were no significant differences between two groups in week of age and body weight (P>0.05).

Results

①The TNBC nude mice with transplanted tumor models were constructed successfully in both two groups, and the tumor formation rates both were 100%. The growth of transplantation tumor in experimental group was relatively slower and the tumor volume was smaller than those of control group. The weight of transplantation tumor in experimental group was (0.562±0.032) g, which was significantly heavier than that of (1.202±0.012) g in control group, and the difference was statistical significant (t=96.792, P<0.001). ②According to the results of Western blotting assay, the relative expression of E6-AP in experimental group was 11 168±441, which was significantly lower than that of (24 284±636) in control group, and the difference was statistical significant (t=75.737, P<0.001). However, the relative expressions of annexin A2 in experimental group and control group were 8 631±651 and 17 976±510, respectively, and the difference was statistical significant (t=50.515, P<0.001). ③Immunohistochemical assay results indicated that the positive expression rate of annexin A2 of experimental group (33%) was significantly lower than that of control group (58%), and the difference was statistically significant (χ2=5.796, P=0.016). The integral optical density (IOD) value of annexin A2 of experimental group (17 976±239) was notably lower than that of control group (26 565±329), and the difference was statistically significant (t=94.439, P<0.001).

Conclusions

Annexin A2 could inhibit biological behavior of breast cancer in vivo maybe through the mediation of E6-AP. If the expression of annexin A2 decreases, it can inhibit the proliferation and growth of breast cancer cells which may become the new target of diagnosis and treatment for TNBC.

图4 免疫组化法检测两组三阴性乳腺癌裸鼠移植瘤组织膜联蛋白A2表达情况(图4A:对照组;图4B:实验组)(SP染色,高倍镜)
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