犬乳腺肿瘤他莫昔芬耐药细胞中差异表达circRNAs的筛选与分析

doi:10.16431/j.cnki.1671-7236.2023.08.039

Abstract

Abstract: 【Objective】 The purpose of this study was to screen and analyze the role of circRNA in canine mammary gland tumor (CMGT) resistance,and provide a new target for tumor diagnosis and treatment.【Method】 Taking CMGT cell CHMm as the research object,we firstly constructed CMGT tamoxifen (TAM) resistant cell lines by concentration gradient method and concentration maintenance method,then verified its resistance by CCK8,and finally screened differentially expressed circRNAs in drug-resistant cells by high-throughput sequencing technology.GO function and KEGG signaling pathway were used to analyze the biological processes in which it might be involved.【Result】 The results showed that after continuous TAM stimulation,the CHMm morphology changed into long spindle shape,and the half inhibitory concentration (IC₅₀) increased from 4.07 to 13.75 μmol/L,indicating that the drug-resistant cells showed stronger TAM resistance.Sequencing results showed that 46 circRNAs were differentially expressed in drug-resistant cells.GO function analysis involved the processes of tumor extracellular matrix,protein ubiquitination,cell adhesion and cell migration,and the major signaling pathways were Notch,Rap1,ErbB,FoxO and PI3K pathways.【Conclusion】 In this study,the tamoxifen-resistant cell line CHMm^TAM was successfully constructed in CMGT and the resistant cells showed stronger proliferation ability and TAM resistance.The differentially expressed circRNAs in parents and drug-resistant cells were screened in high throughput.It was also found that the generation of drug resistance might affect biological processes such as cell signal transduction,cell ubiquitination and cell metastasis through Notch,Rap1 and ErbB pathways,and ultimately participate in tumor drug resistance.

Key words: canine mammary gland tumor; circRNA; tamoxifen-resistant; Notch pathway

CLC Number:

S859.7

XU Enshuang, YANG Chunxue, SUN Yue, TIAN Xue, ZHENG Jiasan, LIU Yun. Screening and Analysis of circRNAs Differentially Expressed in Tamoxifen-resistant Canine Mammary Gland Tumor Cells[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3413-3420.

References

[1] ZHENG H H,DU C T,YU C,et al.Epidemiological investigation of canine mammary tumors in mainland China between 2017 and 2021[J].Frontiers in Veterinary Science,2022,9:843390.
[2] CLARKE R,TYSON J J,DIXON J M.Endocrine resistance in breast cancer-An overview and update[J].Molecular and Cellular Endocrinology,2015,418:220-234.
[3] RONDON-LAGOS M,VILLEGAS V E,RANGEL N,et al.Tamoxifen resistance:Emerging molecular targets[J].International Journal of Molecular Sciences, 2016,17:1357.
[4] SANGER H L,KLOTZ G,RIESNER D,et al.Viroids are single-stranded covalently closed circular RNA molecules existing as highly base-paired rod-like structures[J].Proceedings of the National Academy of Sciences of the United States of America,1976,73:3852-3856.
[5] GUO X Q,HUA Y M.Circular RNAs:Novel regulators of resistance to systemic treatments in breast cancer[J].Neoplasma,2022,69:1019-1028.
[6] TANG Y Y,ZHAO P,ZOU T N,et al.Circular RNA hsa_circ_0001982 promotes breast cancer cell carcinogenesis through decreasing miR-143[J].DNA and Cell Biology,2017,36:901-908.
[7] 曾朝涛,李祖勇,杨正飞.CircRNA-miRNA-mRNA网络在心肺复苏后大鼠心肌损伤中的作用[J].岭南急诊医学杂志,2022,27(6):501-505. ZENG C T,LI Z Y,YANG Z F.Role of circRNA-miRNA-mRNA network in myocardial injury after cardiopulmonary resuscitation in rats[J].Lingnan Journal of Emergency Medicine,2022,27(6):501-505.(in Chinese)
[8] MACEWEN E G.Spontaneous tumors in dogs and cats:Models for the study of cancer biology and treatment[J].Cancer and Metastasis Reviews,1990,9:125-136.
[9] DORN C R,TAYLOR D O,SCHEIDER R,et al.Survey of animal neoplasms in alameda and contra costa counties,California.Ⅱ.Cancer morbidity in dogs and cats from Alameda county[J].Journal of the National Cancer Institute,1968,40:307-318.
[10] BRIEST S,STEARNS V.Tamoxifen metabolism and its effect on endocrine treatment of breast cancer[J].Clinical Advances in Hematology&Oncology,2009,7:185-192.
[11] 张智滔,赵帅.托瑞米芬联合他莫昔芬辅助治疗晚期乳腺癌的效果及对患者生存质量的影响[J].临床合理用药杂志,2022,15(28):96-98. ZHANG Z T,ZHAO S.Effect of torimiphen combined with tamoxifen in adjuvant treatment of advanced breast cancer and its effect on quality of life of patients[J].Chinese Journal of Clinical Rational Drug Use,2022,15(28):96-98.(in Chinese)
[12] XU E,HU M,GE R,et al.LncRNA-42060 regulates tamoxifen densitivity and tumor development via regulating the miR-204-5p/SOX4 axis in canine mammary gland tumor cells[J].Frontiers in Veterinary Science,2021,8:654694.
[13] CHANG C Y,CHOU P P,CHEN W J,et al.Assessment of the tumorigenesis and drug susceptibility of three new canine mammary tumor cell lines[J].Research in Veterinary Science,2010,88:285-293.
[14] OSTRANDER E A.Both ends of the leash-The human links to good dogs with bad genes[J].New England Journal of Medicine,2012,367:636-646.
[15] ALVAREZ C E.Naturally occurring cancers in dogs:Insights for translational genetics and medicine[J].Ilar Journal,2014,55:16-45.
[16] XU E,GUAN T,HU M,et al.Establishment and transcriptome characterization of tamoxifen-resistant canine mammary gland tumor cells[J]. Research in Veterinary Science,2022,145:135-146.
[17] TAVASOLY A,GOLSHAHI H,REZAIE A,et al.Classification and grading of canine malignant mammary tumors[J].Veterinary Research Forum,2013,4:25-30.
[18] 任晓丽,范玉营,皇甫和平,等.表观遗传调控机制在犬肿瘤中的研究进展[J].中国畜牧兽医,2021,48(11):4319-4326. REN X L,FAN Y Y,HUANGFU H P,et al.Research progress of epigenetic regulation mechanism in canine tumor[J].China Animal Husbandry&Veterinary Medicine,2021,48(11):4319-4326.(in Chinese)
[19] DAWOUD A,IHAB ZAKARIA Z,HISHAM RASHWAN H,et al.Circular RNAs:New layer of complexity evading breast cancer heterogeneity[J].Noncoding RNA Research,2023,8:60-74.
[20] KRISHNA B M,JANA S,SINGHAl J,et al.Notch signaling in breast cancer:From pathway analysis to therapy[J].Cancer Letters,2019,461:123-131.
[21] GUO S,LIU M,GONZALEZ-PEREZ R R.Role of Notch and its oncogenic signaling crosstalk in breast cancer[J].Biochimica et Biophysica Acta,2011,1815:197-213.
[22] KONTOMANOLIS E N,KALAGASIDOU S,POULILIOU S,et al.The Notch pathway in breast cancer progression[J].Scientific World Journal,2018,2018:2415489.
[23] ZHANG T,JIANG K,ZHU X,et al.miR-433 inhibits breast cancer cell growth via the MAPK signaling pathway by targeting Rap1a[J].International Journal of Biological Sciences,2018,14:622-632.
[24] 刘娇.Circ_0000231通过miR-140结合RAP1B介导卵巢癌细胞恶性生物学行为及紫杉醇耐药的机制研究[D].沈阳:中国医科大学,2022. LIU J.Circ_0000231 mediated malignant biological behavior and paclitaxel resistance of ovarian cancer cells through miR-140 binding with RAP1B[D].Shenyang:China Medical University,2022.(in Chinese)

Screening and Analysis of circRNAs Differentially Expressed in Tamoxifen-resistant Canine Mammary Gland Tumor Cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 13

Recommended Articles

Metrics

Comments

[1]	CHEN Yanxi, WANG Chen, LUO Yuan, ZHOU Yuancheng, XU Zhiwen, PENG Yuanyi, SONG Zhenhui, LIU Xiao. circRNA Expression Profile Analysis of PK-15 Cells Response to Seneca Virus A Infection [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 280-289.
[2]	LIU Rui, LI Jiangling, DU Huarui, YANG Chaowu, CHEN Jialei, ZHAO Sujun, WANG Qiushi. Identification of Immune-Related circRNA of Thymus in Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2022-2032.
[3]	WANG Dong, LIANG Jiayue, WANG Xiaobing, FENG Juan, ZHANG Ru, XIA Huiyan, XIAO Hongmei. Research Progress of Non-coding RNA in the Development of Hair Follicles of Cashmere Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1817-1827.
[4]	WEI Wenzhuo, ZHENG Chao, WU Yan, LIANG Zhenhua, PI Jingsong, DU Jinping, LI Chengfeng, ZHANG Hao. Characteristic Analysis of circ-ZNF326 in Laying Ducks, Construction of Overexpression Vector and Its Effect on Proliferation of Small Intestinal Epithelial Cells [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 837-844.
[5]	LIU Zhuang, HE Ruyue, CAO Yang, LI Cunyuan, ZHANG Yunfeng, HU Shengwei. Effects of circRNA-Zfp609 on Proliferation and Differentiation of C2C12 Myoblasts [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4535-4544.
[6]	NI Ligang, ZHAO Xuting, WANG Xiaoyan, XU Pan, GAN Yuan, JIN Wen. Screening and Analysis of Differentially Expressed circRNAs in the Lung Tissue of Jiangquhai pigs Infected with Mycoplasma hyopneumoniae [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(6): 2212-2220.
[7]	LIN Zekun, ZHUANG Xiaona, LUO Junyi, CHEN Ting, XI Qianyun, ZHANG Yongliang, SUN Jiajie. Effects of Non-coding RNAs on Skeletal Muscle Development in Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3595-3603.
[8]	LIU Rui, LI Jiangling, CHEN Xiaohui, ZHAO Sujun, WANG Qiushi, HE Zhiping, WANG Yuping. Identification of circRNA Related to Male Reproduction in Pig Testis [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3660-3669.
[9]	XING Baosong, WANG Jing, CHEN Junfeng, REN Qiaoling, MA Qiang, GUO Hongxia, PAN Chuanying, CAO Hai. Effect of miR-199a-5p on Adipogenesis and Prediction and Analysis of Its Target Genes [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 55-63.
[10]	ZHANG Xiao, RAN Xueqin, NIU Xi, HUANG Shihui, WANG Jiafu. Expression and Function Analysis of Circular RNA in Three Tissues of Xiang Pig [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(12): 3870-3881.
[11]	BAI Fengting, LI Lin, CHEN Junhao, ZHAO Bei, PENG Xinyu, WANG Yanhong, FANG Xingtang, ZHANG Chunlei, SONG Chengchuang. Research Progress on Non-coding RNA and Skeletal Muscle Development [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(11): 3584-3594.
[12]	XIE Yueqin, CHEN Ting, LUO Junyi, XI Qianyun, ZHANG Yongliang, SUN Jiajie. Mechanism of circRNA and Its Effect on Development of Animal Muscles [J]. , 2018, 45(8): 2270-2275.
[13]	ZHENG Yuanyuan, HUI Taiyu, GUO Dan, GUO Suling, WANG Yiru, LIU Yuhao, WANG Zicheng, WANG Yingxuan, HUANG Lifei, BAI Wenlin, WANG Zeying. Research Progress on Cashmere Fineness Candidate Genes Regulating by Non-coding RNA [J]. , 2018, 45(11): 3176-3184.