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

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

doi:10.16098/j.issn.0529-1356.2021.01.007

PDF(3922 KB)

Acta Anatomica Sinica ›› 2021, Vol. 52 ›› Issue (1) : 49-54. DOI: 10.16098/j.issn.0529-1356.2021.01.007

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 Mang¹ HUANG Yu¹ LI Yan-jiao¹ ZHANG Ben-si^1*

Author information +

History +

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

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

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. Inhibition of T lymphocytes secreting EphrinAl-Caspase-3 on proliferation of tumor tissue in nude mice with breast cancer[J]. Acta Anatomica Sinica. 2021, 52(1): 49-54 https://doi.org/10.16098/j.issn.0529-1356.2021.01.007

References

［1］ Willis DS, Kennedy C, Kilbride L. Breast cancer screening in women with learning disabilities: current knowledge and considerations［J］. Brit J Learn Disabil, 2008, 36(3):171-184.

［2］ Chen WQ, Li H, Sun KX, et al. Analysis of the incidence and death of malignant tumors in China in 2014 ［J］. China Cancer, 2018, 27(1):1-14. (in Chinese)

陈万青, 李贺, 孙可欣, 等. 2014 年中国恶性肿瘤发病和死亡分析［J］.中国肿瘤, 2018, 27(1):1-14.

［3］ DeSantis CE, Ma J, Goding Sauer A, et al. Breast cancer statistics, 2017, racial disparity in mortality by state［J］. CA Cancer J Clin, 2017, 67(6): 439-448.

［4］ Holzman LB, Marks RM, Dixit VM. A novel immediate-early response gene of endothelium is induced by cytokines and encodes a secreted protein［J］. Mol Cell Biol, 1990, 10(11):5830-5838.

［5］ Gale NW,Holland SJ,Valenzuela DM, et al. Eph receptors and ligands comprise two major specificity subclasses and are reciprocally compartmentalized during embryogenesis.［J］. Neuron,1996,17(1):9-19.

［6］ Pasquale, Elena B. Eph receptors and ephrins in cancer: bidirectional signalling and beyond［J］. Nat Rev Cancer, 2010, 10(3):165-180.

［7］ Yang LH, Shan ChG, Xu QR, et al. Relationship and research progress of Smac, XIAP, caspase-3 and tumorgenesis ［J］. Hebei Pharmaceutical, 2014(4):583-586. (in Chinese)

杨莉晖, 单春光, 许秋荣,等. Smac、XIAP、caspase-3与肿瘤发生的关系及研究进展［J］. 河北医药, 2014(4):583-586.

［8］ Huang Y, Li YJ, Zhang BS, et al. Ephrina1-caspase-3 targeting inhibition of breast cancer cells in vitro ［J］. Chinese Journal of Anatomy, 2018, 41(6): 646-649. (in Chinese)

黄煜, 李艳娇, 张本斯, 等. EphrinA1-caspase-3 对乳腺癌细胞的体外靶向性抑制作用［J］. 解剖学杂志, 2018, 41(6): 646-649.

［9］ Niu JZh, Wang J. HER2, BRCA1, c-myc in the study of breast cancer ［J］. Acta Anatomica Sinica,2000, 31(S1):5-8. (in Chinese)

牛建昭,王健.HER2、BRCA1、c-myc在乳腺癌研究方面的进展［J］.解剖学报,2000, 31(S1):5-8.

［10］ Mitsiades N, Poulaki V, Kotoula V, et al. Fas/Fas ligand up-regulation and Bcl-2 down-regulation may be significant in the pathogenesis of Hashimoto’s thyroiditis［J］. J Clin Endocrinol Metab, 1998, 83(6):2199-2203.

［11］ Kristiansen G, Winzer KJ, Mayordomo E, et al. CD24 expression is a new prognostic marker in breast cancer［J］. Clin Cancer Res, 2003, 9(13):4906-4913.

［12］ Choi YL, Lee SH, Kwon GY, et al. Overexpression of CD24: association with invasiveness in urothelial carcinoma of the bladder［J］. Arch Pathol Lab Med, 2007, 131(2):275-281.

［13］ Surowiak P, Materna V, Kaplenko I, et al. Unfavorable prognostic value of CD24 expression in sections from primary and relapsed ovarian cancer tissue［J］. Int J Gynecol Cancer, 2010, 16(2):515-521.

［14］ Zhao LM, Zhao Sh, Bi RJ, et al. Expression and significance of caspase-3 and caspase-8 in breast cancer ［J］. Chinese Journal of Gerontology, 2017, 37(14):3486-3488. (in Chinese)

赵丽敏, 赵帅, 毕仁杰,等. 乳腺癌组织中caspase-3及caspase-8的表达及意义［J］. 中国老年学杂志, 2017, 37(14):3486-3488.

［15］ Kataoka H, Igarashi H, Kanamori M, et al. Correlation of EPHA2 overexpression with high microvessel count in human primary colorectal cancer［J］. Cancer Sci, 2004, 95(2):136-141.

［16］ Kikawa KD, Vidale DR, Van RE, et al. Regulation of the EphA2 kinase by the low molecular weight tyrosine phosphatase induces transformation.［J］. J Biol Chem, 2002, 277(42):39274-39279.

［17］ Zhang BS, Li Zh, Yang G, et al. Expression and significance of EphA2 and Ephrin A1 in breast cancer in Dali bai region, yunnan province ［J］. Journal of Anatomy, 2017, 40(5):529-534. (in Chinese)

张本斯, 李庄, 杨桂,等. 云南大理白族地区乳腺癌EphA2和Ephrin A1的表达及其意义［J］. 解剖学杂志, 2017, 40(5):529-534.

［18］ Zhang BS, Bian SY, Pan Y, et al. Expression of EphA2 and EphrinA1 in breast cancer and their relationship with clinicopathological factors ［J］. Chinese Journal of Clinical Anatomy, 2017, 35(1):43-47. (in Chinese)

张本斯, 卞思源, 潘云,等. 乳腺癌EphA2和EphrinA1的表达及与临床病理因素的关系［J］. 中国临床解剖学杂志, 2017, 35(1):43-47.

［19］ Xing ZhB, Wang FM, Wang BG, et al. Advances of establishment application of human breast carcinoma model in nude mice ［J］. Modern Oncology,2017,25(23):3889-3892. (in Chinese)

邢志博,王凤梅,王炳高，等.人乳腺癌裸鼠移植模型建立与应用进展［J］.现代肿瘤医学,2017,25(23):3889-3892.

［20］ Guo HR, Wang XY, He Sh, et al. Effect of RNAi targeted silencing on CD105 and Ki67 gene expression in OVCAR3 cells of human ovarian cancer ［J］. Acta Anatomica Sinica,2016,47(4):502-506. (in Chinese)

郭海荣,王晓燕,贺帅,等.RNAi靶向沉默对人卵巢癌OVCAR3细胞CD105和Ki67基因表达的影响［J］.解剖学报,2016,47(4):502-506.

［21］ Yerushalmi R, Woods R, Ravdin PM, et al. Ki67 in breast cancer: prognostic and predictive potential［J］. Lancet Oncol, 2010, 11(2):174-183.

［22］ Tobin NP, Lindstrm LS, Carlson JW, et al. Multi-level gene expression signatures, but not binary, outperform Ki67 for the long term prognostication of breast cancer patients［J］. Mol Oncol, 2014, 8(3):741-752.

［23］ Yao LSh, Zhang BG, Meng K, et al. Correlation and clinical significance of expression of Ki67, CXCR4 and TGF- in elderly breast cancer tissues ［J］. Journal of Applied Gerontology, 2010, 24(5):383-386. (in Chinese)

姚乐申, 张保国, 孟奎,等. Ki67、CXCR4和TGF-β在老年乳腺癌组织中表达的相关性研究及其临床意义［J］. 实用老年医学, 2010, 24(5):383-386.

［24］ Denkert C, Loibl S, Müller BM, et al. Ki67 levels as predictive and prognostic parameters in pretherapeutic breast cancer core biopsies: a translational investigation in the neoadjuvant GeparTrio trial［J］. Ann Oncol, 2013, 24(11):2786-2793.

［25］ Li HZh, Yu K, Wang ZM, et al. relationship between the expression and mutation of related genes in breast cancer ［J］. Acta Anatomica Sinica,2003(4):390-394. (in Chinese)

李红智,俞康,王宗敏,谢丽微.乳腺癌细胞动力学指标与相关基因表达、突变的关系［J］.解剖学报,2003(4):390-394.

［26］ Yerushalmi R, Woods R, Ravdin PM, et al. Ki67 in breast cancer: prognostic and predictive potential［J］. Lancet Oncol, 2010, 11(2):174-183.