|
|
|
| Effects of Transcription Factor ID1 on Proliferation and Apoptosis of Sheep (Ovis aries) Follicular Granulosa Cells |
| ZHANG Pei-Ying, SONG Peng-Yan, ZHOU Rong-Yan*, FU Qiang, ZHOU Ying, DING Hong |
| College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, China |
|
|
|
|
Abstract Inhibitor of DNA binding 1 (ID1) is widely present in the body, and affects cell proliferation and apoptosis. To reveal the role of transcription factor ID1 in ovarian follicular granulosa cells of sheep (Ovis aries), the eukaryotic overexpression vector pcDNA3.1-ID1 was constructed and ID1 RNA interference fragment (siID1) was designed and synthesized. The expression level of ID1 gene was detected by qRT-PCR after 24 h of isolated and cultured ovarian follicular granulosa cells treated with follicle-stimulating hormone (FSH). The results showed that the expression of ID1 gene was extremely significantly decreased after FSH treatment (P<0.01). CCK-8 assay was used to detect the proliferation of granulosa cells within 5 d after transfection with pcDNA3.1, pcDNA3.1-ID1, siNC and siID1 in granulosa cells. After 48 h of transfection, JC-1 assay was used to detect the apoptosis of granulosa cells and the expression levels of proliferation and apoptosis genes were detected by qRT-PCR. The result showed that the survival rate of ID1 overexpressed granulosa cells was reduced, the green fluorescence of JC-1 was enhanced, the expression levels of proliferation-related genes cyclin dependent kinase 2 (CDK2)、cyclin dependent kinase 4 (CDK4) and cyclin D2 (CCND2) were significantly down-regulated (P<0.05), the expression of apoptosis-related gene B-cell lymphoma-2 (Bcl2) was significantly decreased (P<0.05), Bcl2-associated X protein (Bax) gene expression was significantly increased (P<0.05), and caspase-3 (Casp3) gene expression was extremely significantly increased (P<0.01). The higher survival rate of granulosa cells and the lower green fluorescence of JC-1 were found in cells with inhibited ID1 expression. The expression levels of proliferation-related genes CDK2 and CCND2 were significantly increased (P<0.01), CDK4 gene was significantly increased (P<0.05), the expression of Bcl2 was significantly increased (P<0.01), and the expression of Bax was significantly decreased (P<0.05). In summary, transcription factor ID1 inhibited the proliferation and promoted the apoptosis of sheep granulosa cells. This study provides a basis for exploring the biological functions of ID1 in follicular granulosa cells and revealing its role in ovarian follicle development.
|
|
Received: 22 April 2022
|
|
|
|
Corresponding Authors:
* rongyanzhou@126.com
|
|
|
|
[1] 蔡循, 陈国强, 陈竺, 等. 2001. 线粒体跨膜电位与细胞凋亡[J].生物化学与生物物理进展, 28(01): 3-6.
(Cai X, Chen G Q, Chen Z, et al. 2001. Mitochondrial trans-membrane potential and cell apoptosis[J]. Progress in Biochemistry and Biophysics, 28(01): 3-6.)
[2] 陈贝贝, 于丽波, 孙文洲, 等. 2015. 沉默 Id1 基因表达对卵巢癌细胞生长影响的实验研究[J]. 现代肿瘤医学, 23(14): 1959-1962.
(Chen B B, Yu L B, Sun W Z, et al. 2015. The study of ovarian cancer of Id1 gene expres-sion in vivo and in vitro[J]. Journal of Modern Oncology,23(14): 1959-1962.)
[3] 迭小红, 罗庆, 毕杨, 等. 2015. siRNA 沉默 Id1 基因对小鼠骨肉瘤细胞增殖、迁移及凋亡的影响[J]. 中华小儿外科杂志, 36(03): 220-225.
(Die X H, Luo Q, Bi Y, et al. 2015. Effect of Id1 knockdown by small interfering RNA on cell proliferation, migration and apoptosis of mouse osteosarcoma cells[J]. Chinese Journal of Pediat-ric Surgery, 36(03): 220-225.)
[4] 董姿杏, 仇超, 康权, 等. 2017. 靶向抑制 Id1 基因调控 Wnt 信号通路促进骨肉瘤 MG63 细胞凋亡[J]. 肿瘤, 37(09):909-916.
(Dong Z X, Qiu C, Kang Q, et al. 2017. Inter-ference of Id1 gene expression promotes apoptosis of os-teosarcoma MG63 cells by regulating Wnt signaling pathway[J]. Tumor, 37(09): 909-916.)
[5] 蒋秀敏, 刘雨生, 许波. 2015. miR-483-5p 通过靶基因 ERK1 调控人类颗粒细胞增殖凋亡平衡[J]. 安徽医科大学学报, 50(11): 1639-1644.
(Jiang X M, Liu Y S, Xu B.2015. Characterization of miRNA-483-5p anfd targeted gene ERK1 in the regulation proliferation-apoptosis bal-ance of human granulosa cells[J]. Acta Universitatis Me-dicinalis Anhui, 50(11): 1639-1644.)
[6] 李曼曼, 于昊, 薛洋, 等. 2020.生殖激素对山羊颗粒细胞孕酮合成的影响[J]. 中国兽医学报, 40(8):1652-1659.
(Li M M, Yu H, Xue Y, et al. 2020. Effects of reproductive hormones on progesterone synthesis in granulosa cells of goat[J]. Chinese Journal of Veterinary Science, 40(8):1652-1659.)
[7] 李晓军, 秦浚川. 2004. 细胞分化抑制因子(Id)研究进展[J].生物化学与生物物理进展, (10): 865-869.
(Li X J, Qin J C.2004. Progress in cellular inhibitor of differentiation (Id)[J]. Progress in Biochemistry and Biophysics, (10): 865-869.)
[8] 李暄, 佟俊硕, 张大崇, 等. 2021. miRNA 调节卵泡颗粒细胞凋亡及其机制的研究进展[J]. 中国畜牧兽医, 48(12):4429-4441.
(Li X, Tong J S, Zhang D C, et al. 2021. Re-search advances on miRNA in regulating the apoptosis of follicular granulosa cells and its mechanism[J]. China Animal Husbandry & Veterinary Medicine, 48(12):4429-4441.)
[9] 李友炳, 刘豫瑞, 庄则豪. 2004. D 蛋白与细胞分化、细胞周期调控及肿瘤的发生[J]. 福建医科大学学报, (03):358-360.
(Li Y B, Liu Y R, Zhuang Z H. 2004. D pro-tein is involved in cell differentiation, cell cycle regulation and tumorigenesis[J]. Journal of Fujian Medical University, (03): 358-360.)
[10] 夏曦, 王红. 2015. Id1 和 E2-2 参与血管内皮损伤修复机制的研究进展[J]. 医学综述, 21(13): 2308-2310.
(Xia X, Wang H.2015. Research progress in mechanism of Id1 and E2-2 repair of injured involved endothelial[J]. Medi-cal Recapitulate, 21(13): 2308-2310.)
[11] 徐庚全, 樊江峰, 杨世华, 等. 2015. FSH、LH 对体外培养的牦牛卵泡颗粒细胞凋亡及 E2、P 分泌功能的影响[J]. 畜牧兽医学报, 46(06): 932-939.
(Xu G Q, Fan J F, Yang S H, et al. 2015. Effects of FSH, LH on apoptosis and E2, P secretion of yak's granulosa cells cultured in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 46(06):932-939.)
[12] 赵文文, 朱婧颖, 韩颖颖. 2014. CCND 基因家族成员分子进化分析[J]. 医学分子生物学杂志, 11(3): 144-149.
(Zhao W W, Zhu J Y, Han Y Y.2014. Molecular evolution analysis of CCND gene family[J]. Journal of Medi-cal Molecular Biology, 11(3): 144-149.)
[13] Algeciras-Schimnich A, Barnhart B C, Peter M E.2002. Apoptosis-independent functions of killer Caspases[J]. Current Opinion in Cell Biology, 14(6): 721-726.
[14] Baxa D M, Luo X X, Yoshimura F K.2005. Genistein induces apoptosis in T lymphoma cells via mitochondrial dam-age[J]. Nutrition and Cancer, 51(1): 93-101.
[15] Brunelle J K, Letai A.2009. Control of mitochondrial apopto-sis by the Bcl-2 family[J]. Journal of Cell Science, 122(Pt4): 437-441.
[16] Chen D, Forootan S S, Gosney J R, et al. 2014. Increased ex-pression of Id1 and Id3 promotes tumorigenicity by en-hancing angiogenesis and suppressing apoptosis in small cell lung cancer[J]. Genes & Cancer, 5(5-6): 212-225.
[17] Chen Z, Lei L, Wen D, et al. 2019. Melatonin attenuates pal-mitic acid-induced mouse granulosa cells apoptosis via endoplasmic reticulum stress[J]. Journal of Ovarian Re-search, 12(1): 43.
[18] Clark B J, Stocco D M.2009. Expression of the steroidogenic acute regulatory (StAR) protein: A novel LH-induced mitochondrial protein required for the acute regulation of steroidogenesis in mouse leydig tumor cells[J]. Endo-crine Research, 21(1-2): 251-257.
[19] Communal C, Sumandea M, Tombe P D, et al. 2002. Functional consequences of caspase activation in cardiac my-ocytes[J]. Proceedings of the National Academy of Sci-ences of the USA, 99(9): 6252-6252.
[20] Epping J J.2001. Oocyte control of ovarian follicular develop-ment and function in mammals[J]. Reproduction, 122(6): 829.
[21] Hogg K, Etherington S L, Young J M, et al. 2010. Inhibitor of differentiation (Id) genes are expressed in the steroido-genic cells of the ovine ovary and are differentially regu-lated by members of the transforming growth factor-be-ta family[J]. Endocrinology, 151(3): 1247-1256.
[22] Irusta G, Parborell F, Peluffo M, et al. 2003. Steroidogenic acute regulatory protein in ovarian follicles of gonado-tropin-stimulated rats is regulated by a gonadotropin-re-leasing hormone agonist[J]. Biology of Reproduction, 68(5): 1577-1583.
[23] King K L, Cidloeski J A.2003. Cell cycle regulation and apop-tosis[J]. Annual Review of Physiology, 60(1): 601-617. Kolupaeva V, Basilico C. 2012. Overexpression of cyclin E/
[24] CDK2 complexes overcomes FGF-induced cell cycle ar-rest in the presence of hypophosphorylated Rb proteins[J]. Cell Cycle, 11(13): 2557-2566.
[25] Kroemer G, Reed J C.2001. Mitochondrial control of cell death[J]. Nature Medicine, 1(5): 513-519.
[26] Laulier C, Lopez B S.2012. The secret life of Bcl-2: Apopto-sis-independent inhibition of DNA repair by Bcl-2 fami-ly members[J]. Mutation Research-Reviews in Mutation Research, 751(2): 247-257.
[27] Li C Y, Jiang S, Yue Y J, et al. 2012. Effect and pathway of Id1 on the cell growth of nasopharyngeal carcinoma[J]. Medical Journal of Chinese People's Liberation Army, 37(6): 468-471.
[28] Liu L, Zhang Z, Xing D.2011. Cell death via mitochondrial apoptotic pathway due to activation of Bax by lysosom-al photodamage[J]. Free Radical Biology and Medicine, 51(1): 53-68.
[29] Mern D S, Hasskarl J, Burwinkel B.2010. Inhibition of Id proteins by a peptide aptamer induces cell-cycle arrest and apoptosis in ovarian cancer cells[J]. British Journal of Cancer, 103(8): 1237-1244.
[30] Norton J D.2000. ID helix-loop-helix proteins in cell growth, differentiation and tumorigenesis[J]. Journal of Cell Sci-ence, 113(Pt 22): 3897-905.
[31] Ola M S, Nawaz M, Ahsan H.2011. Role of Bcl-2 family pro-teins and caspases in the regulation of apoptosis[J]. Mo-lecular & Cellular Biochemistry, 351(1-2): 41-58. Ruznova M B, Benezra R. 2003. Id proteins in development, cell cycle and cancer[J]. Trends in Cell Biology, 8(13): 410-418.
[32] Satriyo P B, Su C M, Huang W C, et al. 2021. 4-Acetylantro-quinonol B downregulates CDK2/CDK4 expression and DNA damage response signaling for triggers pro-grammed cell death in triple negative breast cancer cells[J]. Toxicology and Applied Pharmacology, 422: 115493.
[33] Shin D H, Park J H, Lee J Y, et al. 2015. Overexpression of Id1 in transgenic mice promotes mammary basal stem cell activity and breast tumorigenesis[J]. Oncotarget, 6(19): 17276-17290.
[34] Su Y, Gao L, Teng L, et al. 2013. Id1 enhances human ovarian cancer endothelial progenitor cell angiogenesis via PI3K/Akt and NF- κB/MMP-2 signaling pathways[J]. Journal of Translational Medicine, 11(1): 132.
[35] Tang X R, Ma L Z, Guo S, et al. 2020. High doses of FSH in-duce autophagy in bovine ovarian granulosa cells via the AKT/mTOR pathway[J]. Reproduction in Domestic Animals, 56(2): 324-332.
[36] Tan H Y, Wang N, hen Y T, et al. 2020. ID1 overexpression in-creases gefitinib sensitivity in non-small cell lung can-cer by activating RIP3/MLKL-dependent necroptosis[J]. Cancer Letters, 475(C): 109-118.
[37] Yao P P, Sheng M J, Weng W H, et al. 2019. High glucose causes apoptosis of rabbit corneal epithelial cells involv-ing activation of PERK-eIF2α-CHOP-caspase-12 signal-ing pathway[J]. International Journal of Ophthalmology, 12(12): 1815-1822.
[38] Bi Y L, Yang S Y, Wang H Y, et al. 2021. Follicle-stimulating hormone is expressed in ovarian follicles of chickens and promotes ovarian granulosa cell proliferation[J]. Journal of Integrative Agriculture, 20(10): 9.
[39] Zhao Y, Luo A, Li S, et al. 2016. Inhibitor of differentiation/ DNA binding 1 (ID1) inhibits etoposide-induced apopto-sis in a c-Jun/c-Fos-dependent manner[J]. Journal of Bi-ological Chemistry, 291(13): 6831-4682. |
|
|
|
