Effect of ciglitazone on adipogenic transdifferentiation of bovine skeletal muscle satellite cells

( Junfang Zhang ),( Qiang Li ),( Yan Yan ),( Bin Sun ),( Ying Wang ),( Lin Tang ),( Enze Wang ),( Jia Yu ),( Kim Margarette Corpuz Nogoy ),( Xiangzi Li ),( Seong-Ho Choi ) 한국축산학회 2021 한국축산학회지 Vol.63 No.4

Ciglitazone is a member of the thiazolidinedione family, and specifically binds to peroxisome proliferator-activated receptor-γ (PPARγ), thereby promoting adipocyte differentiation. We hypothesized that ciglitazone as a PPARγ ligand in the absence of an adipocyte differentiation cocktail would increase adiponectin and adipogenic gene expression in bovine satellite cells (BSC). Muscle-derived BSCs were isolated from six, 18-month-old Yanbian Yellow Cattle. The BSC were cultured for 96 h in differentiation medium containing 5 μM ciglitazone (CL), 10 μM ciglitazone (CM), or 20 μM ciglitazone (CH). Control (CON) BSC were cultured only in a differentiation medium (containing 2% horse serum). The presence of myogenin, desmin, and paired box 7 (Pax7) proteins was confirmed in the BSC by immunofluorescence staining. The CL, CM, and CH treatments produced higher concentrations of triacylglycerol and lipid droplet accumulation in myotubes than those of the CON treatment. Ciglitazone treatments significantly increased the relative expression of PPARγ, CCAAT/enhancer-binding protein alpha (C/EBPα), C/EBPβ, fatty acid synthase, stearoyl-CoA desaturase, and perilipin 2. Ciglitazone treatments increased gene expression of Pax3 and Pax7 and decreased expression of myogenic differentiation-1, myogenin, myogenic regulatory factor-5, and myogenin-4 (p < 0.01). Adiponectin concentration caused by ciglitazone treatments was significantly greater than CON (p < 0.01). RNA sequencing showed that 281 differentially expressed genes (DEGs) were found in the treatments of ciglitazone. DEGs gene ontology (GO) analysis showed that the top 10 GO enrichment significantly changed the biological processes such as protein trimerization, negative regulation of cell proliferation, adipocytes differentiation, and cellular response to external stimulus. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that DEGs were involved in the p53 signaling pathway, PPAR signaling pathway, biosynthesis of amino acids, tumor necrosis factor signaling pathway, non-alcoholic fatty liver disease, PI3K-Akt signaling pathway, and Wnt signaling pathway. These results indicate that ciglitazone acts as PPARγ agonist, effectively increases the adiponectin concentration and adipogenic gene expression, and stimulates the conversion of BSC to adipocyte-like cells in the absence of adipocyte differentiation cocktail.

Ciglitazone, in Combination with All trans Retinoic Acid, Synergistically Induces PTEN Expression in HL-60 Cells

Lee, Seung-ho,Park, Chul-Hong,Kim, Byeong-Su 충남대학교 형질전환복제돼지연구센터 2007 논문집 Vol. No.10

Peroxisome proliferator-activated receptor-gamma(PPARγ)는 DNA와 결합하기 위해 retinoid-X receptor(RXR)와 heterodimer를 형성해야만 한다. 그리고 전사에 대한 최대활성은 수용체에 대한 리간드 특이성에 의하는 것으로 생각되고 있다. 활성화된 PPARγ와 PPARγ 리간드는 종양억제 PTEN의 조절을 통해 종양세포의 성장에 영향으 끼치게 된다. 본 연구의 목적은 PPARγligand, ciglitazone그리고 RXR ligand로 동시에 자극하였을 때 급성전골수성백혈병(APL)세포에 대해 이들이 함께 PTEN upregulate를 조절할 수 있는지를 결정하기 위함이다. 그리고 이들 세포의 성장과 분화주기에 대해 강력한 억제 능이 있는지를 결정하고자 하였다. 즉, 사람의 백혈병세포주인 HL-60세포에 all-trans-retinol과 ciglutazone을 노출시킨 뒤 PTEN 발현에 대한 측정을 위해 RT-PCR법으로 PTEN mRNA 발현 정도를 확인하고 western blot으로 분석하였다. 세포주기의 분석은 propidium iodide(PI) 염색법과 FACScan으로 분석하였고, HL-60 cells에서 PPARγligand, ciglitazone, 그리고 RXR lignd, retinoic acid 그리고 upregulated PTEN 발현에 대한 time-and dose-dependent 방법으로 각각 확인하였던 바 ciglitazone과 retinoic acid를 동시 조합하여 처치하였을 때 유의적인 효과를 인정할 수 있었다. 더욱이 이들 혼합 물질은 세포의 성장과 G₁phase를 동시 억제하는 능력이 있었다. 그러므로 PPARγ의 활성에 있어 RXR heterodimer가 사람의 백혈병세포에 대한 조절 경로로서 존재하며, PTEN의 upregulation을 통해 백혈병을 조절하기 때문에 백혈병의 예방 및 치료 접근에 PPARγ와 RXR ligands가 중요한 역할을 할 것이다. Peroxisome proliferator-activated receptor-gamma (PPARγ) must form a heterodimer with the retinoid-X receptor (RXR) to bind DNA, and its transcriptional activity is thought to be maximized by ligands specific for either receptor. Activated PPARγ and PPARγ ligands may influence tumor growth through regulation of the tumor suppressor PTEN. Our aim in this study was to determine whether co-stimulation with the PPARγ ligand, ciglitazone, and RXR ligand can synergistically upregulate PTEN in human acute promyelocytic leukemia (APL) cells and consequently potentate the inhibition of cell growth and cell cycle progression of these cells. Human leukemia cell line, HL-60 cells were exposed to all-trans-retinol and ciglutazone. The PTEN expression was measured as the level of PTEN mRNA expression by RT-PCR and as the level of PTEN expression by western blot analysis. Cell cycle analysis was carried out by a propidium iodide (PI) staining method and analyzed with a FACScan. The PPARγ ligand, ciglitazone, and the RXR ligand, retinoic acid, upregulated PTEN expression by HL-60 cells in time- and dose-dependent manners, respectively. This was significantly enhanced by a combination of both ciglitazone and retinoic acid. Moreover, these compounds synergistically induced arrests of both cell growth and the G₁phase of the cell cycle. Thus, the activation of the PPARγ: RXR heterodimer may represent a regulatory pathway for human leukemia cells and there may be important roles for PPARγ and RXR ligands in prophylactic and therapeutic approaches for controlling leukemia through the upregulation of PTEN.

PPAR-gamma agonist, ciglitazone, increases pigmentation and migration of human melanocytes

Lee, Joong Sun,Choi, You Mi,Kang, Hee Young Munksgaard 2007 Experimental dermatology Vol.16 No.2

<P>Abstract: </P><P>Peroxisome proliferator-activated receptors (PPARs) play an important role in cellular responses. It was reported that three subtypes of PPAR are expressed in human melanocytes. In this study, we investigated the effects of the PPAR-<I>&ggr;</I> agonist, ciglitazone, on pigmentation and migration of human melanocytes. Ciglitazone stimulated the melanin content of cells and cultured skin. This increase in pigmentation was due to the stimulation of tyrosinase activity and expression of tyrosinase and microphthalmia-associated transcription factor protein of the melanocytes. Migration was increased after ciglitazone treatment, which was observed by the Boyden chamber checkerboard analysis and a scratch motility assay. These results suggest the regulatory role of PPAR-<I>&ggr;</I> in pigmentation and migration of human melanocytes.</P>

Ciglitazone, in Combination with All trans Retinoic Acid, Synergistically Induces PTEN Expression in HL-60 Cells

Lee, Seung-Ho,Park, Chul-Hong,Kim, Byeong-Su 한국식품위생안전성학회 2006 한국식품위생안전성학회지 Vol.21 No.3

Inhibition of ERK activity enhances the cytotoxic effect of peroxisome proliferator-activated receptor γ (PPARγ) agonists in HeLa cells

Chang, Ha Kyun,Kim, Dae Seong,Chae, Jung Jun,Kim, Minsun,Myong, Jun-Pyo,Lee, Keun Ho,Lee, Myoung Woo,Park, Tae Chul Elsevier 2017 Biochemical and biophysical research communication Vol. No.

<P><B>Abstract</B></P> <P>In this study, we examined whether the peroxisome proliferator-activated receptor γ (PPARγ) agonists, ciglitazone (CGZ) and troglitazone (TGZ), induce cell death in human cervical cancer HeLa cells. The cells were treated with a range of CGZ or TGZ doses for 24 or 48 h. Low concentrations of CGZ (≤10 μM) or TGZ (≤20 μM) had no effect on cell viability whereas higher doses induced cell death in a time- and dose-dependent manner as evidenced by the detection of activated caspase-3 and PARP cleavage. Treatment with the PPARγ antagonist GW9662 followed by PPARγ agonists did not increase CGZ- or TGZ-induced cell death, indicating that PPARγ agonists induced HeLa cell death independently of PPARγ. Moreover, ERK1/2 activation was observed at a CGZ concentration of 25 μM and a TGZ concentration of 35 μM, both of which induced cell death. To elucidate the role of ERK1/2 activated by the two PPARγ agonists, the effect of U0126, an inhibitor of ERK1/2, on PPARγ-agonist-induced cell death was examined. Treatment with 10 or 20 μM U0126 followed by CGZ or TGZ induced the down-regulation of ERK1/2 activity and a decrease in Bcl-2 expression accompanied by the collapse of mitochondrial membrane potential, which in turn significantly enhanced CGZ- or TGZ-induced apoptotic cell death. Our results suggest that PPARγ agonists are capable of inducing apoptotic cell death in HeLa cells independently of PPARγ and that inhibition of ERK1/2 activity offers a strategy to enhance the cytotoxicity of PPARγ agonists in the treatment of cervical cancer.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The PPARγ agonists CGZ and TGZ induce apoptotic cell death in HeLa cells. </LI> <LI> CGZ or TGZ induces apoptotic cell death independently of PPARγ in HeLa cells. </LI> <LI> Inhibition of ERK1/2 enhances CGZ- or TGZ-induced cell death via the collapse of MMP. </LI> </UL> </P>