title: CYP1A1 and CYP1A2 expression levels are differentially regulated in three-dimensional spheroids of liver cancer cells compared to two-dimensional monolayer cultures.
abstract: Compared to two-dimensional (2D) monolayer cultures, three-dimensional (3D) tumor cell culture models are thought to be structurally more similar to the in vivo tumor microenvironment. We investigated the regulation of the expression of genes encoding the drug-metabolizing enzymes CYP1A1 and CYP1A2 in 3D spheroids comprised of cells of the human hepatocellular carcinoma cell JHH1, Huh7, and HepG2. Expression of CYP1A1 and CYP1A2 in the spheroids was higher than that in 2D cultured cells. Expression of CYP1A1 and CYP1A2 is regulated by aryl hydrocarbon receptor (AhR) in 2D cultured cells. Knockdown of AhR in spheroids suppressed CYP1A1 expression; however, CYP1A2 expression levels remained unchanged. Moreover, we found that pregnane X receptor (PXR) likely regulated CYP1A2 expression in JHH1, HepG2, and Huh7 spheroids and that CYP1A1 expression in JHH1 and Huh7 3D spheroids is regulated not only by AhR but also by PXR. It is well known that gene expression levels are different between 3D spheroids and 2D monolayer cultured cells, and our results indicate that the regulation of gene expression also varies between the two culture conditions. Taken together, these results underlie a novel finding regarding the regulation of drug-metabolizing enzyme expression in liver cancer cells growing as 3D spheroids.
authors.affiliation: Pharmacodynamics and Molecular Genetics, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuda, Yahaba-CHO, Siwa-Gun, Iwate 028-3694, Japan. Electronic address: jterashi@iwate-med.ac.jp.
authors.forename: Jun
authors.initials: J
authors.lastname: Terashima
authors.affiliation: Pharmacodynamics and Molecular Genetics, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuda, Yahaba-CHO, Siwa-Gun, Iwate 028-3694, Japan; Department of Pharmacy, Iwate Medical University Hospital, 19-1 Uchimaru, Morioka, Iwate 020-8505, Japan.
authors.forename: Shinpei
authors.initials: S
authors.lastname: Goto
authors.affiliation: Pharmacodynamics and Molecular Genetics, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuda, Yahaba-CHO, Siwa-Gun, Iwate 028-3694, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Hattori
authors.affiliation: Pharmacodynamics and Molecular Genetics, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuda, Yahaba-CHO, Siwa-Gun, Iwate 028-3694, Japan.
authors.forename: Sawaka
authors.initials: S
authors.lastname: Hoshi
authors.affiliation: Pharmacodynamics and Molecular Genetics, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuda, Yahaba-CHO, Siwa-Gun, Iwate 028-3694, Japan.
authors.forename: Midori
authors.initials: M
authors.lastname: Ushirokawa
authors.affiliation: Department of Pharmacy, Iwate Medical University Hospital, 19-1 Uchimaru, Morioka, Iwate 020-8505, Japan.
authors.forename: Kenzo
authors.initials: K
authors.lastname: Kudo
authors.affiliation: Pharmacodynamics and Molecular Genetics, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuda, Yahaba-CHO, Siwa-Gun, Iwate 028-3694, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Habano
authors.affiliation: Pharmacodynamics and Molecular Genetics, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuda, Yahaba-CHO, Siwa-Gun, Iwate 028-3694, Japan.
authors.forename: Shogo
authors.initials: S
authors.lastname: Ozawa
mesh_terms: D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics / Q000378:metabolism*; D019388:Cytochrome P-450 CYP1A2 / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000201:enzymology* / Q000235:genetics; D000077297:Pregnane X Receptor; D034622:RNA Interference; D018336:Receptors, Aryl Hydrocarbon / Q000235:genetics / Q000378:metabolism; D011987:Receptors, Steroid / Q000235:genetics / Q000378:metabolism; D018874:Spheroids, Cellular; D014162:Transfection
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D019363:Cytochrome P-450 CYP1A1; D019388:Cytochrome P-450 CYP1A2; D000077297:Pregnane X Receptor; D018336:Receptors, Aryl Hydrocarbon; D011987:Receptors, Steroid; C111235:AHR protein, human; C569463:CYP1A1 protein, human; C504573:CYP1A2 protein, human
keywords: 3D culture; Aryl hydrocarbon receptor; CYP1A1; CYP1A2; Liver cancer; Pregnane X receptor; Spheroid