title: Interference RNA-based silencing of endogenous SMAD4 in porcine granulosa cells resulted in decreased FSH-mediated granulosa cells proliferation and steroidogenesis.
abstract: FSH plays a critical role in granulosa cell (GC) proliferation and steroidogenesis through modulation by factors including bone morphogenetic proteins family, which belongs to transforming growth factor β (TGFB) superfamily. TGFBs are the key factors in maintaining cell growth and differentiation in ovaries. However, the interaction of FSH and TGFB on the GCs' proliferation and steroidogenesis remains to be elucidated. In this study, we have investigated the role of SMAD4, a core molecule mediating the intracellular TGFB/SMAD signal transduction pathway, in FSH-mediated proliferation and steroidogenesis of porcine GCs. In this study, SMAD4 was knocked down using interference RNA in porcine GCs. Our results showed that SMAD4-siRNA causes specific inhibition of SMAD4 mRNA and protein expression after transfection. Knockdown of SMAD4 significantly inhibited FSH-induced porcine GC proliferation and estradiol production and changed the expression of cyclin D2, CDK2, CDK4, CYP19a1, and CYP11a1. Thus, these observations establish an important role of SMAD4 in the regulation of the response of porcine GCs to FSH.
authors.affiliation: College of Animal Science and Technology, Nanjing Agricultural University, Weigang 1, Nanjing 210095, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Xia
authors.initials: X
authors.lastname: Chen
authors.forename: Xinxiu
authors.initials: X
authors.lastname: Li
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Haiyan
authors.initials: H
authors.lastname: Zhang
authors.forename: Yu
authors.initials: Y
authors.lastname: He
authors.forename: Jingjing
authors.initials: J
authors.lastname: Wang
authors.forename: Yongyan
authors.initials: Y
authors.lastname: Zhao
authors.forename: Baole
authors.initials: B
authors.lastname: Zhang
authors.forename: Yinxue
authors.initials: Y
authors.lastname: Xu
mesh_terms: D000818:Animals; D001141:Aromatase / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism; D015536:Down-Regulation; D004958:Estradiol / Q000096:biosynthesis*; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism*; D018816:Genes, cdc; D006107:Granulosa Cells / Q000378:metabolism*; D011374:Progesterone / Q000096:biosynthesis; D034622:RNA Interference; D011962:Receptors, FSH / Q000378:metabolism; D015398:Signal Transduction; D051901:Smad4 Protein / Q000235:genetics / Q000378:metabolism*; D013552:Swine; D014162:Transfection
chemical_list: D011962:Receptors, FSH; D051901:Smad4 Protein; D011374:Progesterone; D004958:Estradiol; D005640:Follicle Stimulating Hormone; D001141:Aromatase; D002786:Cholesterol Side-Chain Cleavage Enzyme