Br‐DIF‐1 accelerates dimethyl sulphoxide‐induced differentiation of P19CL6 embryonic carcinoma cells into cardiomyocytes

• Published in: British journal of pharmacology Vol. 165; no. 4; pp. 870 - 879
• Main Authors: Seya, K, Kanemaru, K, Matsuki, M, Hongo, K, Kitahara, H, Kikuchi, H, Oshima, Y, Kubohara, Y, Okumura, K, Motomura, S, Furukawa, K‐I
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.02.2012; Nature Publishing Group
• Subjects: Animals; beating; Biological and medical sciences; Br‐DIF‐1; Calcium Channel Blockers - pharmacology; Calcium Channels, T-Type - physiology; cardiomyocyte differentiation; Cell Differentiation - drug effects; Cell Line, Tumor; Dimethyl Sulfoxide; dimethyl sulphoxide; Gene Expression - drug effects; Hexanones - pharmacology; Medical sciences; mibefradil; Mibefradil - pharmacology; Mice; mouse; Myocytes, Cardiac - cytology; Myocytes, Cardiac - drug effects; Myocytes, Cardiac - physiology; P19CL6; Pharmacology. Drug treatments; Research Papers; siRNA; T‐type Ca2+ channel; Prostaglandin-endoperoxide synthase; Embryo; RNA interference; T-type Ca; Calcium antagonist; channel; dimethyl sulphoxide; Br-DIF-1; Tumor cell; cardiomyocyte differentiation; Mibefradil; P19CL6; Enzyme; beating; T-type calcium channel; Rodentia; Enzyme inhibitor; siRNA; Non steroidal antiinflammatory agent; Gene silencing; Vertebrata; Mammalia; Mouse; Benzimidazole derivatives; Animal; Cardiomyocyte; Dimethyl sulfoxide; Antihypertensive agent; Oxidoreductases; Differentiation; Heart cell
• ISSN: 0007-1188; 1476-5381
• DOI: 10.1111/j.1476-5381.2011.01541.x

BACKGROUND AND PURPOSE Stem cell transplantation therapy is a promising option for treatment of severe ischaemic heart disease. Dimethyl sulphoxide (DMSO) differentiates P19CL6 embryonic carcinoma cells into cardiomyocyte‐like cells, but with low differentiation capacity. To improve the degree of this differentiation, we have assessed several derivatives of the differentiation‐inducing factor‐1 (DIF‐1), originally found in the cellular slime mould Dictyostelium discoideum, on P19CL6 cells. EXPERIMENTAL APPROACH P19CL6 cells were cultured with each derivative and 1% DMSO for up to 16 days. Differentiation was assessed by measuring the number of beating and non‐beating aggregates, and the expression of genes relevant to cardiac tissue. The mechanism of action was investigated using a T‐type Ca2+ channel blocker. KEY RESULTS Of all the DIF‐1 derivatives tested only Br‐DIF‐1 showed any effects on cardiomyocyte differentiation. In the presence of 1% DMSO, Br‐DIF‐1 (0.3–3 µM) significantly and dose‐dependently increased the number of spontaneously beating aggregates compared with 1% DMSO alone, by day 16. Expression of mRNA for T‐type calcium channels was significantly increased by Br‐DIF‐1 + 1% DMSO compared with 1% DMSO alone. Mibefradil (a T‐type Ca2+ channel blocker; 100 nM) and a small interfering RNA for the T‐type Ca2+ channel both significantly decreased the beating rate of aggregates induced by Br‐DIF‐1 + 1% DMSO. CONCLUSIONS AND IMPLICATIONS Br‐DIF‐1 accelerated the differentiation, induced by 1% DMSO, of P19CL6 cells into spontaneously beating cardiomyocyte‐like cells, partly by enhancing the expression of the T‐type Ca2+ channel gene.