敲低SQLE基因对奶牛乳腺上皮细胞增殖及凋亡的影响

doi:10.16431/j.cnki.1671-7236.2020.08.019

Abstract

Abstract: In order to explore the effect of SQLE gene on the apoptosis and proliferation of dairy mammary gland cells in vitro,the relative expression of SQLE gene,apoptosis and cycle-related genes of dairy mammary epithelial cells with SQLE gene knockdown were detected by Real-time quantitative PCR.Cells were screened by cycle,apoptosis detection kits and flow cytometry.The results showed that the relative expression of SQLE gene was significantly decreased after siRNA (SQLE siRNA) transfection.The cell proliferation was significantly inhibited (P<0.05).The number of cells in G1 phase significantly decreased (P<0.05),and the proportion of cells in S phase showed an increasing trend (P<0.05).No significant difference in cells in G2 phase.The relative expression of apoptotic gene Fas and proliferation and cycle related genes P27 significantly increased (P<0.05), Cyclin D1,Bcl-2,P21 and Bax genes decreased significantly (P<0.05).In summary,knockdown of SQLE gene could inhibit the proliferation of breast epithelial cells by regulating the expression of related genes.

Key words: cow; SQLE gene; mammary epithelial; proliferation; apoptosis

CLC Number:

Q786

HU Xiangwei, YANG Shuai, YUAN Jing, CHEN Chao, YANG Liguo. Effects of SQLE Gene Knock-down on Proliferation and Apoptosis of Dairy Cow Mammary Epithelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(8): 2510-2517.

References

[1] SEIKI S,FRISHMAN W H.Pharmacologic inhibition of squalene synthase and other downstream enzymes of the cholesterol synthesis pathway:A new therapeutic approach to treatment of hypercholesterolemia[J].Cardiology in Review,2009,17(2):70-76.
[2] GILL S,STEVENSON J,KRISTIANA I,et al.Cholesterol-dependent degradation of squalene monooxygenase,a control point in cholesterol synthesis beyond HMG-CoA reductase[J].Cell Metabolism,2011,13(3):260-273.
[3] HOUTEN S M,SCHNEIDERS M S,WANDERS R J,et al.Regulation of isoprenoid/cholesterol biosynthesis in cells from mevalonate kinase-deficient patients[J].Journal of Biological Chemistry,2003,278(8):5736-5743.
[4] WANDERS R J,ROMEIJN G J.Cholesterol biosynthesis,peroxisomes and peroxisomal disorders:Mevalonate kinase is not only deficient in Zellweger syndrome but also in rhizomelic chondrodysplasia punctata[J].Journal of Inherited Metabolic Disease,1998,21(3):309-312.
[5] LIU D,WONG C C,FU L,et al.Squalene epoxidase drives NAFLD-induced hepatocellular carcinoma and is a pharmaceutical target[J].Science Translational Medicine,2018,10(437):eaap9840.
[6] SHI Y,JIANG,H.Discovery of a potent HMG-CoA reductase degrader that eliminates statin-induced reductase accumulation and lowers cholesterol[J].Nature Communications,2018,9(1):1-13.
[7] SUI Z H,ZHOU J H,CHENG Z J,et al.Squalene epoxidase (SQLE) promotes the growth and migration of the hepatocellular carcinoma cells[J].Tumor Biology,2015,36(8):6173-6179.
[8] 李奎,康相涛,刘英,等.固始鸡免疫器官内细胞凋亡基因Fas和FasL的动态表达[J].中国农业科学,2008,41(5):1489-1496. LI K,KANG X T,LIU Y,et al.Dynamic expression of apoptosis genes Fas and FasL in immune organs of Gushi chickens[J].Scientia Agricultura Sinica,2008,41(5):1489-1496.(in Chinese)
[9] 高薇,侯微,李伟,等.细胞凋亡机制研究进展[J].中国畜牧兽医,2014,41(10):150-156. GAO W,HOU W,LI W,et al.Research progress on cell apoptosis mechanism[J].China Animal Husbandry & Veterinary Medicine,2014,41(10):150-156.(in Chinese)
[10] 张清峰.Fas/FasL系统调控公牛精子发生的研究进展[J].中国畜牧兽医,2009,36(8):99-102. ZHANG Q F.The research progress of Fas/FasL system expands bull spermatogenesis[J].China Animal Husbandry & Veterinary Medicine,2009,36(8):99-102.(in Chinese)
[11] HNIT SS,XIE C,YAO M,et al.p27 Kip1 signaling:Transcriptional and post-translational regulation[J].International Journal of Biochemistry & Cell Biology,2015,68:9-14.
[12] VALLONTHAIEL A G,SINGH M K,DINDA A K,et al.Prognostic significance of cytoplasmic p27 in oral squamous cell carcinoma[J].Journal of Oral Pathology & Medicine,2016,45(7):475-480.
[13] CHEN G,CHENG Y,ZHANG Z,et al.Prognostic significance of cytoplasmic p27 expressioin human melanoma[J].Cancer Epidemiology Biomarkers Prevention,2011,20(10):2212-2221.
[14] LIU Y,TEMPLETON D M.Iron-loaded cardiac myocytes stimulate cardiac myofibroblast DNA synthesis[J].Molecular & Cellular Biochemistry,2006,281(1-2):77-85.
[15] WU W J,LIU Q Q,LIU Y X,et al.Dixdc1 targets Cyclin D1 and p21via PI3K pathway activation to promote Schwann cell proliferation after sciatic nerve crush[J].Biochemical & Biophysical Research Communications,2016,478(2):956-963.
[16] LIAO K,LI J,WANG Z.Dihydroartemisinin inhibits cell proliferation via AKT/GSK3beta/cyclin D1 pathway and induces apoptosis in A549 lung cancer cells[J].International Journal of Clinical & Experimental Pathology,2014,7(12):8684-8691.
[17] CHEN A,HUANG X,XUE Z,et al.The role of p21 in apoptosis,proliferation,cell cycle arrest,and antioxidant activity in UVB-irradiated human HaCaT keratinocytes[J].Medical Science Monitor Basic Research,2015,21:86-95.
[18] KVANSAKUL M,HINDS MG.The Bcl-2 family:Structures,interactions and targets for drug discovery[J].Apoptosis,2015,20(2):136-150.
[19] ESKES R,ANTONSSON B,OSEN-SAND A,et al.Bax-induced cytochrome C release from mitochondria is independent of the permeability transition pore but highly dependent on Mg²⁺ ions[J].The Journal of Cell Biology,1998,143(1):217-224.
[20] VYSSOKIKH MY,ZOROVA L,ZOROV D,et al.Bax releases cytochrome c preferentially from a complex between porin and adenine nucleotide translocator.Hexokinase activity suppresses this effect[J].Molecular Biology Reports,2002,29(1-2):93-96.
[21] MAHDAVI S,KHODARAHMI P,ROODBARI N H.Effects of cadmium on Bcl-2/Bax expression ratio in rat cortex brain and hippocampus[J].Human & Experimental Toxicology,2018,37(3):321-328.
[22] ZHANG M Q,GAO J L,LIAO X D,et al.miR-454 regulates triglyceride synthesis in bovine mammary epithelial cells by targeting PPAR-γ[J].Gene,2019,691(7):1-7.
[23] SEAGROVES T N,LYDON J P,HOVEY R C,et al.C/EBPbeta (CCAAT/enhancer binding protein) controls cell fate determination during mammary gland development[J].Molecular Endocrinology,2000,14(3):359-368.
[24] COSTA A,LOPEZ-VILLALOBOS N,SNEDDON N W,et al.Invited review:Milk lactose-current status and future challenges in dairy cattle[J].Journal of Dairy Science,2019,102(7):5883-5898.
[25] GENG Y,YU Q,SICINSKA E,et al.Deletion of the p27Kip1 gene restores normal development in cyclin D1-deficient mice[J].Proceedings of the National Academy of Sciences of the United States of America,2001,98(1):194-199.
[26] INMAN J L,ROBERTSON C,MOTT J D,et al.Mammary gland development:Cell fate specification,stem cells and the microenvironment[J].Development,2015,142(6):1028-1042.
[27] ZAVIZION B,POLITIS I,GOREWIT R C.Bovine mammary myoepithelial cells.1.isolation,culture,and characterization[J].Journal of Dairy Science,1992,75(12):3367-3380.
[28] TONG H L,LI Q Z,GAO X J,et al.Establishment and characterization of a lactating dairy goat mammary gland epithelial cell line[J].Vitro Cellular & Developmental Biology Animal,2012,48(3):149-155.
[29] 王洋,金一.siRNA及其运载系统研究进展[J].中国畜牧兽医,2017,44(5):1281-1288. WANG Y,JIN Y.Research progress on small interfering RNA and its delivery systems[J].China Animal Husbandry & Veterinary Medicine,2017,44(5):1281-1288.(in Chinese)
[30] 丁旭娜,王凤龙,丁玉林,等.奶牛乳腺上皮细胞和成纤维细胞的体外培养与鉴定[J].黑龙江畜牧兽医,2012,5:1-5. DING X N,WANG F L,DING Y L,et al.Identification and in vitro culture of bovine mammary epithelial cells and fibroblast[J].Heilongjiang Animal Science and Veterinary Medicine,2012,5:1-5.(in Chinese)
[31] ZHANG J Y,ZHANG F,QUN C,et al.Critical protein GAPDH and its regulatory mechanisms in cancer cells[J].Cancer Biology & Medicine,2015,12(1):10-22.
[32] QIN Y,ZHANG Y,TANG Q,et al.SQLE induces epithelial-to-mesenchymal transition by regulating of miR-133b in esophageal squamous cell carcinoma[J].Acta Biochimica et Biophysica Sinica,2017,49(2):138-148.
[33] KIM J H,KIM C N,KANG D W.Squalene epoxidase correlates E-cadherin expression and overall survival in colorectal cancer patients:The impact on prognosis and correlation to clinicopathologic features[J].Journal of Clinical Medicine,2019,8(5):632.
[34] MAHONEY C E,PIRMAN D,CHUBUKOV V,et al.A chemical biology screen identifies a vulnerability of neuroendocrine cancer cells to SQLE inhibition[J].Nature Communications,2019,10:96.

Effects of SQLE Gene Knock-down on Proliferation and Apoptosis of Dairy Cow Mammary Epithelial Cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHONG Yueyun, YANG Shuzhan, CHEN Feiyue, LU Zhier, LI Bingxin, LI Wanyan, TIAN Yunbo, HUANG Yunmao, XU Danning, CAO Nan. Effects of LPS on Apoptosis of Goose Embryo-derived Hepatocytes ROS/p38/MAPK Signaling Pathway [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2224-2232.
[2]	XIAO Peng, SHANG Jianghua, YANG Chunyan, LI Mengqi, DUAN Anqin, MA Xiaoya, FENG Chao, HUANG Chenqian, ZHANG Bo, ZHOU Jinchen, WEI Kelong, ZHENG Wei, ZHENG Haiying. Effects of Alpha-linolenic Acid on Buffalo Ovarian Granulosa Cells Cultured in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2380-2387.
[3]	LI Can, LI Chuchu, ZENG Wei, ZHANG Ning, JI Chunxiao, CHEN Tao. Recombinant Expression and Function Study of RegⅢγ Protein in Porcine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2414-2426.
[4]	FAN Yimeng, WEI Yuanyuan, WANG Huiru, GA yu, ZHANG Yannan, ZHAO Qingyu, HAO Zhihui. Potential Mechanisms of Chinese Veterinary Medicine Compound Rukang Granules in the Treatment of Dairy Cow Mastitis Based on Network Pharmacology and Molecular Docking Technology [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2507-2517.
[5]	HU Honglian, LI Chao, CHEN Liqing, SONG Liwen, ZHAO Meng, ZHANG Jianxia, WANG Dian, GAO Min. Effects of Dietary β-carotene Supplementation on Lactation Performance, Blood Routine,Serum Biochemical and Immune Indexes of Dairy Cows during the Lactating Peak Period [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1375-1384.
[6]	GUO Yaru, LI Kexin, WANG Li, LIU Jinyu, SHI Lei, REN Youshe. Effects of Selenium on Lead-Induced Apoptosis of Mouse Leydig Cells in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1404-1411.
[7]	LI Dan, ZHANG Haisen, WANG Yiqun, GAO Dengke, ZHAO Hongcong, LI Chao, JIN Yaping, CHEN Huatao. Eukaryotic Expression Vector Construction,Bioinformatics and Tissue Expression Profiles Analysis of CRY1 Gene in Dairy Cows [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 870-881.
[8]	KONG Weiyi, WANG Feifei, YUE Kang, QIN Junjie, ZHU Hao, GUO Yansheng. Effect of Guiqi Yimu Oral Liquid on Plasma Metabolites in Postpartum Dairy Cows Based on UPLC-MS/MS Metabolomics Technology [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 913-923.
[9]	HUANG Xiaoqian, YUAN Junzhao, LIU Jinhe, ZOU Hui, GU Jianhong, YUAN Yan, LIU Xuezhong, LIU Zongping, BIAN Jianchun. Protective Effect of Honokiol on Cadmium-induced Renal Injury in Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1007-1015.
[10]	ZHEN Zhen, WANG Mei, WANG Yimin, HU Debao, ZHANG Linlin, LI Xin, GUO Yiwen, GUO Hong, DING Xiangbin. Effect of RNA Methylation Transfer Enzyme METTL3 on the Proliferation and Myogenic Differentiation of Bovine Skeletal Muscle Satellite Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1025-1036.
[11]	ZHONG Huachen, WANG Lifang, GUO Chenyang, LIU Jialin, SONG Jie. Isolation,Identification and Drug Resistance Analysis of Pathogenic Bacteria from Milk Samples and Environment of Mastitis in Inner Mongolia [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 817-826.
[12]	HE Yue, WANG Hui, CHEN Ruipeng, YU Zhixue, TANG Xiangfang, GUO Yuming, XIONG Benhai. Research Progress on Optical and Electrochemical Biosensors for the Diagnosis of Important Diseases in Dairy Cows [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 827-837.
[13]	TIAN Shuyue, SHEN Yingchao, WANG Xisheng, WANG Min, YI Minna, Tseweendolmaa, TAO Li, ZHAO Bilig, Manglai, Gerelchimeg. Study on Establishment of Immortalized Cell System of Equine Fetal Fibroblasts in Mongolian Horse [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 46-57.
[14]	PAN Siyao, ZHANG Dandan, LIU Quanhui, WANG Guodong, LYU Danwei, HUANG Ben. Effects of Inhibition of miR-142-3p on Lactation Function of Chemical Induced Mammary Epithelial Cells in Dairy Goats [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 58-66.
[15]	ZENG Zhiqin, LOU Wenqi, XU Jingyi, MA Yehua, ZHANG Yongfu, REN Xiaoli, CHEN Shaokan, YAN Lei, GUO Gang, ZHANG Zhen, WANG Yachun. Analysis of Impact Factors and Phenotype Prediction for Transforming Progesterone Concentration in Chinese Holstein Cows During Lactation [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 186-197.