TCAP knockdown by RNA interference inhibits myoblast differentiation in cultured skeletal muscle cells.
Null mutation of titin-cap (TCAP) causes limb-girdle muscular dystrophy type 2G (LGMD2G). LGMD2G patients develop muscle atrophy, and lose the ability to walk by their third decade. Previous findings suggest that TCAP regulates myostatin, a key regulator of muscle growth. We tested the hypothesis that TCAP knockdown with RNA interference will lead to differential expression of genes involved in muscle proliferation and differentiation, impairing muscle cell growth. mRNA from cultured cells treated with TCAP siRNA duplex constructs was analyzed using Northern blots and real-time RT-PCR. siRNA treatment decreased TCAP mRNA expression in differentiating muscle cells. Significant (p<0.05) decreases in mRNA were observed for myogenic regulatory factors. siRNA treatment also prevented development of the normal phenotype of muscle cells. Our findings suggest that TCAP knockdown with RNA interference alters normal muscle cell differentiation.
Markert
Chad D
CD
Department of Neurology and Wake Forest Institute for Regenerative Medicine, Wake Forest University Health Sciences, Medical Center Boulevard, Winston-Salem, NC 27157, USA.
Ning
Jie
J
Staley
Jerry T
JT
Heinzke
Laura
L
Childers
Charles K
CK
Ferreira
J Andries
JA
Brown
Marybeth
M
Stoker
Aaron
A
Okamura
Carol
C
Childers
Martin K
MK
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D049109:Cell Proliferation; D064211:Connectin; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000235:genetics / Q000502:physiology*; D049288:Muscular Dystrophies, Limb-Girdle / Q000235:genetics / Q000473:pathology; D017570:MyoD Protein / Q000235:genetics; D032448:Myoblasts, Skeletal / Q000166:cytology / Q000378:metabolism*; D018008:Myogenin / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D064211:Connectin; D009124:Muscle Proteins; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; C503801:Myog protein, mouse; D018008:Myogenin; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C577294:Tcap protein, mouse; D007335:Insulin-Like Growth Factor II