title: hCG-induced endoplasmic reticulum stress triggers apoptosis and reduces steroidogenic enzyme expression through activating transcription factor 6 in Leydig cells of the testis.
abstract: Endoplasmic reticulum (ER) stress generally occurs in secretory cell types. It has been reported that Leydig cells, which produce testosterone in response to human chorionic gonadotropin (hCG), express key steroidogenic enzymes for the regulation of testosterone synthesis. In this study, we analyzed whether hCG induces ER stress via three unfolded protein response (UPR) pathways in mouse Leydig tumor (mLTC-1) cells and the testis. Treatment with hCG induced ER stress in mLTC-1 cells via the ATF6, IRE1a/XBP1, and eIF2α/GADD34/ATF4 UPR pathways, and transient expression of 50 kDa protein activating transcription factor 6 (p50ATF6) reduced the expression level of steroidogenic 3β-hydroxysteroid dehydrogenase Δ5-Δ4-isomerase (3β-HSD) enzyme. In an in vivo model, high-level hCG treatment induced expression of p50ATF6 while that of steroidogenic enzymes, especially 3β-HSD, 17α-hydroxylase/C17-20 lyase (CYP17), and 17β-hydrozysteroid dehydrogenase (17β-HSD), was reduced. Expression levels of steroidogenic enzymes were restored by the ER stress inhibitor tauroursodeoxycholic acid (TUDCA). Furthermore, lentivirus-mediated transient expression of p50ATF6 reduced the expression level of 3β-HSD in the testis. Protein expression levels of phospho-JNK, CHOP, and cleaved caspases-12 and -3 as markers of ER stress-mediated apoptosis markedly increased in response to high-level hCG treatment in mLTC-1 cells and the testis. Based on transmission electron microscopy and H&E staining of the testis, it was shown that abnormal ER morphology and destruction of testicular histology induced by high-level hCG treatment were reversed by the addition of TUDCA. These findings suggest that hCG-induced ER stress plays important roles in steroidogenic enzyme expression via modulation of the ATF6 pathway as well as ER stress-mediated apoptosis in Leydig cells.
authors.affiliation: Center for Food and Nutritional Genomics Research, Kyungpook National University, Daegu, Republic of Korea.
authors.forename: Sun-Ji
authors.initials: SJ
authors.lastname: Park
authors.forename: Tae-Shin
authors.initials: TS
authors.lastname: Kim
authors.forename: Choon-Keun
authors.initials: CK
authors.lastname: Park
authors.forename: Sang-Hee
authors.initials: SH
authors.lastname: Lee
authors.forename: Jin-Man
authors.initials: JM
authors.lastname: Kim
authors.forename: Kyu-Sun
authors.initials: KS
authors.lastname: Lee
authors.forename: In-Kyu
authors.initials: IK
authors.lastname: Lee
authors.forename: Jeen-Woo
authors.initials: JW
authors.lastname: Park
authors.forename: Mark A
authors.initials: MA
authors.lastname: Lawson
authors.forename: Dong-Seok
authors.initials: DS
authors.lastname: Lee
mesh_terms: D015096:3-Hydroxysteroid Dehydrogenases / Q000235:genetics / Q000378:metabolism; D051702:Activating Transcription Factor 6 / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D002460:Cell Line; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007985:Leydig Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D015398:Signal Transduction; D013737:Testis / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D056811:Unfolded Protein Response / Q000187:drug effects
chemical_list: D051702:Activating Transcription Factor 6; D006063:Chorionic Gonadotropin; D015096:3-Hydroxysteroid Dehydrogenases; C511890:3beta-hydroxysteroid dehydrogenase 4, mouse