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

doi:10.11843/j.issn.0366-6964.2022.12.032

Abstract

Abstract: 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.

Key words: canine mammary tumor, CHMp cells, THBS1, biological properties, cell apoptosis

CLC Number:

S857.4

WU Juye, LU Baochun, ZHU Yufan, JIA Kun. The Effects of Thrombospondin 1 on the Biological Characteristics of Canine Mammary Tumor Cells in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4470-4481.

References 20

[1]	SALAS Y, MÁRQUEZ A, DIAZ D, et al. Epidemiological study of mammary tumors in female dogs diagnosed during the period 2002-2012:a growing animal health problem[J]. PLoS One, 2015, 10(5):e0127381.
[2]	MARCONATO L, LEO C, GIRELLI R, et al. Association between waste management and cancer in companion animals[J]. J Vet Intern Med, 2009, 23(3):564-569.
[3]	刘春晖, 杨蕾, 孙艳争, 等. 北京地区犬肿瘤发生率调查[J]. 中国兽医杂志, 2013, 49(1):63-64.LIU C H, YANG L, SUN Y Z, et al. Investigation on the incidence of canine tumor in Beijing area[J]. Chinese Journal of Veterinary Medicine, 2013, 49(1):63-64. (in Chinese)
[4]	HUANG T T, SUN L, YUAN X L, et al. Thrombospondin-1 is a multifaceted player in tumor progression[J]. Oncotarget, 2017, 8(48):84546-84558.
[5]	包楠丁, 贾永峰. THBS1在肿瘤中作用的研究进展[J]. 标记免疫分析与临床, 2021, 28(4):703-707.BAO N D, JIA Y F. The research progress of THBS1 in tumors[J]. Labeled Immunoassays and Clinical Medicine, 2021, 28(4):703-707. (in Chinese)
[6]	高献争, 左士宇, 王晓慧, 等. 血小板凝血酶蛋白1下调对肺腺癌A549细胞生物学功能的影响[J]. 中华实验外科杂志, 2018, 35(4):702-705.GAO X Z, ZUO S Y, WANG X H, et al. Effects of thrombospondin 1 down-regulation on biological function of lung adenocarcinoma A549 cells[J]. Chinese Journal of Experimental Surgery, 2018, 35(4):702-705. (in Chinese)
[7]	ROUANNE M, ADAM J, GOUBAR A, et al. Osteopontin and thrombospondin-1 play opposite roles in promoting tumor aggressiveness of primary resected non-small cell lung cancer[J]. BMC Cancer, 2016, 16(1):483.
[8]	吴居业, 陈和壁, 卢宝春, 等. TNC、PREX1、THBS1和SERPINE1的mRNA在犬乳腺肿瘤中表达水平的差异性分析[J]. 中国兽医科学, 2021, 51(5):663-670.WU J Y, CHEN H B, LU B C, et al. Analysis of the differences in the expression levels of the mRNA of TNC, PREX1, THBS1 and SERPINE1 in canine mammary tumors[J]. Chinese Veterinary Science, 2021, 51(5):663-670. (in Chinese)
[9]	HORTA R D S, LAVALLE G E, CUNHA R M D C, et al. Influence of surgical technique on overall survival, disease free interval and new lesion development interval in dogs with mammary tumors[J]. Adv Breast Cancer Res, 2014, 3(2):38-46.
[10]	石慧, 周云, 吴宜, 等. 结肠癌组织中WWOX、THBS1、FOXO1的表达变化及意义[J]. 山东医药, 2020, 60(28):17-20.SHI H, ZHOU Y, WU Y, et al. Expression changes and significance of WWOX, THBS1 and FOXO1 in colon cancer tissues[J]. Shandong Medical Journal, 2020, 60(28):17-20. (in Chinese)
[11]	CARVALHO M I, PIRES I, PRADA J, et al. High COX-2 expression is associated with increased angiogenesis, proliferation and tumoural inflammatory infiltrate in canine malignant mammary tumours:a multivariate survival study[J]. Vet Comp Oncol, 2017, 15(2):619-631.
[12]	PÉREZ ALENZA M D, PENA L, NIETO A I, et al. Clinical and pathological prognostic factors in canine mammary tumors[J]. Ann Ist Super Sanita, 1997, 33(4):581-585.
[13]	李妍, 蓝婷英, 庞博,等. NLRP3炎症小体及其下游炎症因子在犬乳腺肿瘤组织中的表达[J]. 畜牧兽医学报, 2022, 53(4):1252-1258.LI Y, LAN T Y, PANG B, et al. Expression of NLRP3 inflammasome signal associated genes in canine mammary tumors and clinical significance[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1252-1258.(in Chinese)
[14]	PAL S K, NGUYEN C T K, MORITA K I, et al. THBS1 is induced by TGFB1 in the cancer stroma and promotes invasion of oral squamous cell carcinoma[J]. J Oral Pathol Med, 2016, 45(10):730-739.
[15]	DE FRAIPONT F, NICHOLSON A C, FEIGE J J, et al. Thrombospondins and tumor angiogenesis[J]. Trends Mol Med, 2001, 7(9):401-407.
[16]	殷茜, 王佩佩, 彭睿, 等. miR-19a调控血小板反应蛋白在结肠癌淋巴转移中的机制[J]. 华中科技大学学报:医学版, 2019, 48(1):42-47.YIN Q, WANG P P, PENG R, et al. Mechanism of miR-19a regulating thrombospondin in lymphatic metastasis of colon cancer[J]. Acta Medicinae Universitatis Scientiae et Technologiae Huazhong, 2019, 48(1):42-47. (in Chinese)
[17]	KLOPFLEISCH R, GRUBER A D. Derlin-1 and stanniocalcin-1 are differentially regulated in metastasizing canine mammary adenocarcinomas[J]. J Comp Pathol, 2009, 141(2-3):113-120.
[18]	LUTFUL KABIR F M, ALVAREZ C E, BIRD R C. Canine mammary carcinomas:a comparative analysis of altered gene expression[J]. Vet Sci, 2015, 3(1):1.
[19]	LI P, JIANG H L, PENG H, et al. Non-structural protein 5 of Zika virus interacts with p53 in human neural progenitor cells and induces p53-mediated apoptosis[J]. Virol Sin, 2021, 36(6):1411-1420.
[20]	李雪. PHLDB3、P53、Bax、Bcl-2在非小细胞肺癌中的表达及临床意义[D]. 泸州:西南医科大学, 2021.LI X. Expression and clinical significance of PHLDB3, P53, Bax, Bcl-2 in non-small cell lung cancer[D]. Luzhou:Southwest Medical University, 2021. (in Chinese)

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

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 20

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	WANG Wanjie, CHEN Nanzhu, ZOU Huiying, ZHOU Xinyi, HAO Haisheng, PANG Yunwei, ZHU Huabin, ZHAO Xueming, YU Dawei, DU Weihua. Effects of Histone Methyltransferase ASH1L Overexpression on Proliferation and Apoptosis of Bovine Cumulus Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3358-3368.
[2]	DAN Yixin, YANG Lu, XIANG Hua, ZHANG Huanrong, REN Yupeng, YANG Falong, HE Honghong, ZHU Jiangjiang. Effects of BIRC5 on the Cycle and Apoptosis of Goat Testis Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1511-1524.
[3]	LI Dehao, LI Yisui, MA Jingyun, SUN Yuan, ZHANG Xiangbin. Biological Characteristics and Genome Analysis of Lysogenic Salmonella Phage PEA2-3 [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1632-1640.
[4]	REN Xiaoli, FAN Yuying, HUANGFU Heping, LIU Yun, SHI Dongmei. Effect of GSK126 on Epithelial-mesenchymal Transition of Canine Mammary Tumor Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1721-1729.
[5]	XU Huihao, LIU Jiangyu, LI Qijuan, ZHENG Xiaobo, LIN Jiahao, JIN Yipeng, LIN Degui. Expression and Prognosis of Hexokinase 2 in Canine Mammary Tumors [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1310-1324.
[6]	REN Xiaoli, FAN Yuying, HUANGFU Heping, WANG Jun, JIN Shuangxing, LIU Yun, SHI Dongmei. Mechanism of miR-502 Regulating Proliferation, Migration, Invasion and EMT of Canine Mammary Tumor Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4379-4388.
[7]	XU Songwei, XIANG Hua, ZHANG Huanrong, YANG Lu, WANG Xianjun, SALANG Wenzhu, ZHU Jiangjiang. Effects of Interfering SERBP1 on the Cell Proliferation, Cell Cycle and Apoptosis in Goat Turbinate Bone Primary Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2708-2720.
[8]	ZHANG Zhenzhen, LI Jianzeng, JIANG Ruirui, MA Yanchao, CAI Chunxia, ZHANG Lujie, GUO Yujie, JI Jinqing, HAN Lu, TIAN Yadong, KANG Xiangtao, HAN Ruili. Isolation, Culture and Identification of Chicken Chondrocytes and the Effect of Overexpression of miR-15a on Chondrocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1749-1758.
[9]	LI Yaqi, WANG Xianzhong, ZHANG Jiaojiao. Research Progress of Cell Proliferation and Apoptosis Regulated by Non-thermal Plasma and Its Application in Animal Husbandry [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 1-10.
[10]	WANG Junyue, DAI Jianjun, ZHANG Shushan, SUN Lingwei, WU Caifeng, ZHANG Defu. Effects of High Pressure Homogeneous Egg Yolk on Apoptosis of Boar Sperm Cryopreservation [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(8): 2190-2199.
[11]	HE Yirong, ZHANG Yijie, YANG Wei, CHEN Xiaocong, ZHANG Jiaxiang, CHEN Peifu. Isolation, Identification and Biological Properties of a Lytic Phage against Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 763-771.
[12]	ZHANG Keshan, GAO Guangliang, LI Qin, ZHAO Xianzhi, LI Jing, WANG Qigui. The Function of NPFFR1 on Hormone Secretion and Apoptosis of Follicle Granulosa Cell in Geese [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(10): 2822-2831.
[13]	DIAO Hong-xiu, LI Yi, LIN De-gui, ZHANG Di. Expression of MUC1 and Its Clinical Value in Canine Mammary Tumors [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(7): 1511-1516.
[14]	NIE Hai-tao, HUANG Xin-ai, LIU Hao, GUO Yi-xuan, WANG Zi-yu, ZHANG Yan-li, WAN Yong-jie, FAN Yi-xuan, ZHANG Guo-min, WANG Feng, WANG Jie. Effect of Different Nutrient Levels on Follicle Development and Glucose Metabolic Pathway Involved in Folliculogenesis of Hu Sheep during Luteal Phase [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 942-953.
[15]	YAO Yi-long, LI Yin-xia, REHEMAN Anwaier, ZHANG Jun, MENG Chun-hua, WANG Hui-li, QIAN Yong, CAO Shao-xian. Cloning of Sp1 Gene CDS Region of Hu Sheep and Its Effect on Proliferation and Apoptosis of Granulosa Cells [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(11): 2098-2106.