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  

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

作　　者：时娅琪 唐洗敏 李庄 张丽梅 鲁雪静 周凌智 徐芒 黄煜 李艳娇 张本斯 SHI Ya-qi;TANG Xi-min;LI Zhuang;ZHANG Li-mei;LU Xue-jing;ZHOU Ling-zhi;XU Mang;HUANG Yu;LI Yan-jiao;ZHANG Ben-si(Department of Anatomy,College of Basic Medicine,Dali University,Yunnan Dali 671000,China;Department of Thoracic Surgery,Affiliated Hospital of Dali University,Yunnan Dali 671000,China)

机构地区：[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

出　　处：《解剖学报》2021年第1期49-54,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金(81460465);云南省教育厅科研基金(2020Y0539)。

摘　　要：目的探讨分泌Ephrin Al-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Ephrin A1-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。Ephrin A1-Caspase-3治疗组第2天可以检测出Ephrin A1-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Ephrin A1-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Ephrin Al-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Ephrin Al-Caspase-3的表达。结论Ephrin Al-Caspase-3可显著抑制乳腺癌细胞的生长,发挥抗肿瘤效应。Objective To study the inhibitory effect of T lymphocytes secreting Ephrin Al-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 Ephrin A1-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 8 th 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 Ephrin A1-Caspase-3 was detected in the Ephrin A1-Caspase-3 treatment group on the 2 nd day,reached the peak on the 8 th day,and then the secretion decreased gradually.No expression of Ephrin A1-Caspase-3 was detected in the PBS control group and the T lymphocyte group.The presence of dispersed green fluorescent protein-labeled Ephrin Al-Caspa

关 键 词：分子靶向治疗 乳腺癌模型 Ephrin A1-Caspase-3 KI67 酶联免疫吸附测定 裸鼠 

分 类 号：R323.2[医药卫生—人体解剖和组织胚胎学]