解剖学报 ›› 2021, Vol. 52 ›› Issue (1): 49-54.doi: 10.16098/j.issn.0529-1356.2021.01.007

• 肿瘤生物学 • 上一篇    下一篇

分泌EphrinAl-Caspase-3的T淋巴细胞对裸鼠乳腺癌组织生长的抑制作用

时娅琪1 唐洗敏1 李庄2 张丽梅1 鲁雪静2 周凌智2 徐芒1 黄煜1 李艳娇1 张本斯1*   

  1. 1.大理大学基础医学院解剖学教研室; 2.大理大学附属医院胸外科,云南 大理 671000
  • 收稿日期:2019-10-15 修回日期:2020-01-07 出版日期:2021-02-06 发布日期:2021-02-06
  • 通讯作者: 张本斯 E-mail:ben-si-zhang@163.com
  • 基金资助:
    云南白族地区乳腺癌EphA2表达检测以及间充质干细胞为载体的重组免疫毒素靶向治疗的研究

Inhibition of T lymphocytes secreting EphrinAl-Caspase-3 on proliferation of tumor tissue in nude mice with breast cancer

SHI Ya-qi1  TANG Xi-min1  LI Zhuang2  ZHANG Li-mei1  LU Xue-jing2  ZHOU Ling-zhi2  XU Mang HUANG Yu1  LI Yan-jiao1  ZHANG Ben-si1*   

  1. 1.Department of Anatomy, College of Basic Medicine, Dali University, Yunnan Dali671000,China; 2.Department of Thoracic Surgery, Affiliated Hospital of Dali University, Yunnan Dali671000,China
  • Received:2019-10-15 Revised:2020-01-07 Online:2021-02-06 Published:2021-02-06
  • Contact: ZHANG Ben-si E-mail:ben-si-zhang@163.com

摘要:

目的  探讨分泌EphrinAl-Caspase-3的T淋巴细胞体内移植对癌细胞的抑制作用。  方法将裸鼠(n=35) 接种乳腺癌细胞,构建裸鼠乳腺癌模型。待肿瘤体积达到0.1 cm3大小,选取30只具有平均大小瘤组织的裸鼠随机分为PBS组、未感染腺病毒组、感染Ad-EphrinA1-Caspase-3的T淋巴细胞组,经瘤内移植,每隔2~3 d测量肿瘤大小。另选取3组荷瘤裸鼠,经上述细胞移植后,每隔2~3 d获取裸鼠皮下肿瘤组织匀浆,ELISA检测EphrinA1-Caspase-3的含量。实验结束后各组动物断颈处死剥离肿瘤制成切片,在荧光显微镜下观察表达绿色荧光蛋白的T淋巴细胞,免疫荧光法进行Caspase-3、Ki-67的检测。  结果乳腺癌细胞接种裸鼠1周后可用手摸到皮下肿瘤,证明乳腺癌细胞荷瘤动物模型制做成功。接种后第8天各组裸鼠肿瘤体积差异变大,治疗组与PBS组/T淋巴细胞组相比差异极显著(P<0.05)。单纯T淋巴细胞移植组肿瘤体积虽较PBS对照组生长缓慢,但两者间并无明显统计学差异。EphrinA1-Caspase-3治疗组第2天可以检测出EphrinA1-Caspase-3的表达,第8天达到高峰,随后分泌量逐渐降低。PBS对照组和T淋巴细胞组未检测到EphrinA1-Caspase-3的表达。在治疗组肿瘤组织中观察到分散的绿色荧光蛋白标记的EphrinAl-Caspase-3-T淋巴细胞,而在PBS组和T淋巴细胞组中未检测到绿色荧光蛋白的存在。治疗组感染细胞中,Caspase-3阳性细胞比例上调,Ki-67阳性细胞比例下调。在PBS组和T淋巴细胞组未检测到EphrinAl-Caspase-3的表达。  结论EphrinAl-Caspase-3可显著抑制乳腺癌细胞的生长,发挥抗肿瘤效应。

关键词: 分子靶向治疗, 乳腺癌模型, EphrinA1-Caspase-3, Ki67, 酶联免疫吸附测定, 裸鼠

Abstract:

Objective  To study the inhibitory effect of T lymphocytes secreting EphrinAl-Caspase-3 in vivo and on the growth of cancer cells in nude mice with breast cancer.   Methods  Nude mice(n=35)were inoculated with breast cancer cells to construct a nude mouse model of breast cancer. When the tumor volume reached 0.1 cm3, 30 nude mice with average size tumor tissue were randomly divided into PBS group,uninfected adenovirus group, T lymphocyte infected with Ad-EphrinA1-Caspase-3 group, and intratumoral transplantation. Tumor size was measured every day 2 to 3. Three groups of tumor-bearing nude mice were selected. After the above-mentioned cell transplantation, the subcutaneous tumor tissue homogenate was obtained every day 2 to 3, and the content of EphrinA1-Caspase-3 was detected by ELISA. At the end of the experiment, the animals in each group were sacrificed by cervical dissection and sliced. The presence of T lymphocytes expressing green fluorescent protein was observed under a fluorescence microscope, and Caspase-3 and Ki-67 were detected by immunofluorescence.    Results  After one week of inoculation of breast cancer cells into nude mice, the presence of subcutaneous tumors could be touched by hand, which proved that the tumor-bearing animals of breast cancer cells were successfully modeled. On the 8th day after inoculation, the tumor volume of the nude mice in each group became larger, and the difference between the treatment group and the PBS group/T lymphocyte group was extremely significant (P<0.05). Although the tumor volume of the T lymphocyte transplantation group was slower than that of the PBS control group, there was no statistically significant difference between the two. The expression of EphrinA1-Caspase-3 was detected in the EphrinA1-Caspase-3 treatment group on the 2nd day, reached the peak on the 8th day, and then the secretion decreased gradually. No expression of EphrinA1-Caspase-3 was detected in the PBS control group and the T lymphocyte group. The presence of dispersed green fluorescent protein-labeled EphrinAl-Caspase-3-T lymphocytes was observed in the tumor tissues of the treatment group, while the presence of green fluorescent protein was not detected in the PBS group and the T lymphocyte groups. In the infected cells of the treatment group, the proportion of Caspase-3 positive cells was up-regulated, and the proportion of Ki-67 positive cells was down-regulated. No expression of EphrinAl-Caspase-3 was detected in the PBS group and the T lymphocyte group.   ConclusionEphrinAl-Caspase-3 can significantly inhibit the growth of breast cancer cells, thereby exerting an anti-tumor effect.

Key words: Molecular targeted therapy,  , Breast cancer model,  , EphrinA1-Caspase-3,  , Ki67,  , Enzyme linked immunosorbent assay,  , Nude mouse

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