分泌EphrinAl-Caspase-3的T淋巴细胞抑制乳腺癌细胞生长的动物研究

Animal experimental study on the inhibition of breast cancer growth by in vivo transplantation of T lymphocytes secreting EphrinAl-Caspase-3

目的探讨分泌EphrinAl-Caspase-3的T淋巴细胞体内移植对乳腺癌生长的抑制作用。方法采用6周龄的BALB/c裸鼠接种乳腺肿瘤细胞构建裸鼠乳腺癌模型后,按随机数字表法随机分为3组:PBS组瘤内注射10μL PBS,阴性对照组瘤内注射1×10~6个未感染腺病毒的T淋巴细胞,感染组瘤内注射1×10~6个EphrinAlCaspase3-T淋巴细胞,每隔3天用游标卡尺测量肿瘤的大小(0、3、6、9、12及15 d)至实验结束。处死裸鼠取其肿瘤组织检测组织中EphrinAl-Caspase-3的含量,并行病理学检查观察表达绿色荧光蛋白的T淋巴细胞的存在,以及Caspase-3和Ki-67阳性细胞的表达情况。结果第0天和第3天时,3组裸鼠的肿瘤体积比较差异均无统计学意义(P>0.05);第6天及以后,感染组和PBS组/阴性对照组比较差异均有统计学意义(P<0.05),但各时点PBS组和阴性对照组的肿瘤体积比较差异均无统计学意义(P>0.05)。感染组的肿瘤组织中可见散在的绿色荧光蛋白标记的EphrinAl-Caspase-3-T淋巴细胞的存在,而PBS组和阴性对照组均没有检测到绿色荧光蛋白的存在。与PBS组和阴性对照组比较,感染组的肿瘤细胞中,Caspase-3阳性细胞比例较高,Ki-67阳性细胞比例较低。感染组在第3天就可以检测出EphrinAl-Caspase-3的表达,第6天时达到高峰,随后分泌量逐渐降低;PBS组和阴性对照组各时点均没有检测到EphrinAl-Caspase-3的表达。结论 EphrinAl-Caspase-3能明显抑制体内乳腺癌细胞的生长,促进凋亡。

Objective To investigate the inhibitory effect of T lymphocyte transplantation of EphrinAl-Caspase-3 on the growth of breast cancer. Methods Six-week-old BALB/c nude mice were used to inoculate breast cancer cells to construct a nude mouse model of breast cancer. They were randomly divided into 3 groups according to random number table: PBS group received intratumoral injection of 10 μL PBS, and negative control group received intratumoral injection of 1×10~6 T lymphocytes uninfected with adenovirus, 1×10~6 EphrinAl-Caspase3-T lymphocytes were injected intratumorally into the infected group, and the tumors size(0, 3, 6, 9, 12 and 15 d) were measured with vernier calipers every 3 days until end of experiment. The content of EphrinAl-Caspase-3 in the tissues of the nude mice was measured.The presence of T lymphocytes expressing green fluorescent protein and the ratio of Caspase-3-positive and Ki-67-positive cell were observed by pathological examination. Results On the day 0 and day 3, there were no significant difference in tumor volume between the 3 groups(P>0.05). On the 6 th day and later, the difference between the infected group and the PBS group/negative control group were statistically significant(P<0.05), but there were no significant difference in tumor volume between the PBS group and negative control group at each time point(P>0.05). The presence of scattered green fluorescent protein-labeled EphrinAl-Caspase-3-T lymphocytes was observed in the tumor tissues of the infected group, while the presence of green fluorescent protein were not detected in the PBS group and the negative control group. In the infected cells, ratio of Caspase-3-positive cell was up-regulated and ratio of Ki-67-positive cell was down-regulated. The expression of EphrinAl-Caspase-3 could be detected on the 3 rd day in the infected group, and at the peak on the 6-day, then the amount of secretion gradually decreased. The expression of EphrinAl-Caspase-3 were not detected in the PBS group and the negative control group at each time point. Conclusion EphrinAl-Caspase-3 can significantly inhibit the growth of breast cancer cells and promote apoptosis.