血小板凝血酶蛋白1对犬乳腺肿瘤细胞体外生物学特性的影响分析  

The Effects of Thrombospondin 1 on the Biological Characteristics of Canine Mammary Tumor Cells in vitro

作　　者：吴居业 卢宝春 祝宇凡 贾坤[1,2,3] WU Juye;LU Baochun;ZHU Yufan;JIA Kun(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China;Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases,Guangzhou 510642,China;Guangdong Pet Engineering Technology Research Center,Guangzhou 510642,China)

机构地区：[1]华南农业大学兽医学院,广州510642 [2]广东省兽医临床重大疾病综合防控重点实验室,广州510642 [3]广东省宠物工程技术研究中心,广州510642

出　　处：《畜牧兽医学报》2022年第12期4470-4481,共12页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：广州市科技计划项目(202201010033)。

摘　　要：本研究旨在深入探讨血小板凝血酶蛋白1(THBS1)对犬乳腺肿瘤细胞CHMp的影响,并阐明其影响犬乳腺肿瘤发生发展的作用机制。通过构建THBS1慢病毒稳定过表达和THBS1沉默表达的犬乳腺肿瘤细胞CHMp细胞系,采用CCK-8试验、划痕试验、Transwell试验、原位荧光检测和流式细胞术检测THBS1对CHMp细胞增殖、迁移、侵袭、细胞凋亡和细胞周期情况的影响;通过qRT-PCR和Western blot检测THBS1对CHMp细胞凋亡相关因子(p53、Bcl-2、Bax)表达情况的影响,验证THBS1对CHMp细胞凋亡通路的影响。结果显示,过表达THBS1能有效增强犬乳腺肿瘤细胞CHMp的增殖、迁移和侵袭能力,并且减少CHMp细胞的凋亡数量,而沉默THBS1后结果与之相反;流式细胞术得出THBS1能够影响CHMp细胞的细胞周期分布;经qRT-PCR和Western blot检测发现,在THBS1过表达后,p53和Bcl-2的表达量均明显增高,Bax的表达量明显减少,在沉默THBS1后结果与之相反。结果表明,THBS1的异常表达能够影响犬乳腺肿瘤细胞CHMp的增殖、迁移、侵袭以及细胞周期;此外,THBS1能够通过影响细胞凋亡因子p53、Bcl-2和Bax的表达来影响CHMp细胞凋亡相关通路,进而影响犬乳腺肿瘤的发生发展。The aim of this study was to investigate the effect of Thrombospondin-1(THBS1)on canine mammary tumor cell CHMp,and to clarify its mechanism of action on the development of canine mammary tumors.THBS1 lentivirus-overexpressing cell lines and THBS1-silencing cell lines were constructed from canine mammary tumor cell CHMp.Then CCK-8 assay,scratch assay,Transwell assay,in situ fluorescence detection and flow cytometry were used to detect the effect of THBS1 on the proliferation,migration,invasion,apoptosis and cell cycle of CHMp cells.Finally,the effect of THBS1 on the expression of apoptosis-related factors(p53,Bcl-2,Bax)in CHMp cells was detected by qRT-PCR and Western blot,and the effect of THBS1 on the apoptosis pathway of CHMp cell was further verified.The results showed that overexpression of THBS1 can effectively enhance the proliferation,migration and invasion of canine mammary tumor cells CHMp,and reduce the number of apoptotic CHMp cells,while THBS1 silencing results in the opposite;Flow cytometry showed that THBS1 could affect the cell cycle distribution of CHMp cells;qRT-PCR and Western blot detection showed that after THBS1 overexpression,the expression levels of p53 and Bcl-2 were significantly increased,and the expression level of Bax was significantly decreased,and the results were the opposite after silencing THBS1.The results showed that abnormal expression of THBS1 could affect the proliferation,migration,invasion and cell cycle of canine mammary tumor cell CHMp.In addition,THBS1 can affect the related pathways of CHMp apoptosis factors by affecting the expression of apoptosis factors p53,Bcl-2 and Bax,and play an important role in canine mammary tumors,thereby affecting the occurrence and development of canine mammary tumors.

关 键 词：犬乳腺肿瘤 CHMp细胞 THBS1 生物学特性 细胞凋亡 

分 类 号：S857.4[农业科学—临床兽医学]