PKC promotes proliferation of airway smooth muscle cells by regulating cyclinD1 expression in asthmatic rats.
OBJECTIVE
To determine whether protein kinase C (PKC) has any effect on the expression of cyclinD1, a key regulator of growth control and G1/S transition, and to investigate the underlying molecular mechanisms of PKC involving the remodeling of the asthmatic airway smooth muscle (ASM).


METHODS
The treatment of synchronized ASM cells from asthmatic rats with PKC-specific agonist phorbol 12-myristate 13-acetate (PMA) and antagonist 2-{1-[3-(amidinothio) propyl]-1Hindol-3-yl}-3-(1-methylindol-3-yl) maleimide methanesulfonate salt (Ro31-8220) was followed by the proliferation assay. PKCalpha and cyclinD1 expressions in ASM cells (ASMC) were detected by RT-PCR and Western blotting. The relation between PKCalpha and cyclinD1 was assessed by linear regression analysis. The effect of the construct recombinant plasmid pcDNA3.1-antisense cyclinD1 (pcDNA3.1-ascyclinD1) on the proliferation of ASMC was found to be induced by PMA.


RESULTS
The data showed phorbol ester-dependent PKCalpha promoted the proliferation of ASMC. The closely-positive correlation existed between the expression of PKCalpha and cyclinD1 at the transcriptional (r=0.821, P<0.01) and translational (r=0.940, P<0.01) levels. pcDNA3.1-ascyclinD1 could inhibit the proliferation of ASMC. pcDNA3.1-ascyclinD1 almost completely attenuated the PMA-induced proliferation effect as Ro31-8220+pcDNA3.1.


CONCLUSIONS
The proliferation of ASMC by PKC might by regulated by the cyclinD1 expression in asthmatic rats.
Qiao
Li-fen
LF
Department of Respiratory Medicine, Huazhong University of Science and Technology, Wuhan 430030, China.
Xu
Yong-jian
YJ
Liu
Xian-sheng
XS
Xie
Jun-gang
JG
Wang
Jin
J
Du
Chun-ling
CL
Zhang
Jian
J
Ni
Wang
W
Chen
Shi-xin
SX
D000818:Animals; D001249:Asthma / Q000473:pathology*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D019938:Cyclin D1 / Q000096:biosynthesis* / Q000235:genetics; D016172:DNA Fingerprinting; D005786:Gene Expression Regulation; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000473:pathology*; D010957:Plasmids / Q000235:genetics; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011493:Protein Kinase C / Q000502:physiology*; D051381:Rats; D011914:Rats, Inbred BN
D018809:Proliferating Cell Nuclear Antigen; D019938:Cyclin D1; D011493:Protein Kinase C