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[Cyclin D1 is involved in human pulmonary artery smooth muscle cells proliferation and migration induced by cigarette smoke extract.] [Ariticle in Chinese]

XIANG Min, XU Yong-Jian^*, LIU Xian-Sheng, ZENG Da-Xiong

Department of Respiratory Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

Abstract

The present study was aimed to investigate the role of cyclin D1 in human pulmonary artery smooth muscle cells (HPASMCs)proliferation and migration induced by cigarette smoke extract (CSE). The eukaryotic expression vector of antisense cyclin D1 gene(pIRES2-EGFP-ascyclin D1) was recombinated. The recombinant and empty vector were separately transfected into normal HPASMCsusing liposome. Then the cells were treated with or without 5% CSE. The cells were randomly divided into six groups: control group,vector group, antisense cyclin D1 group, 5% CSE group, vector+5% CSE group and antisense cyclin D1+5% CSE group. Theexpressions of cyclin D1 mRNA and protein were detected by real-time fluorescence RT-PCR and Western blot, respectively. Theproliferation of HPASMCs was examined by cell cycle analysis, MTT assay and proliferation cell nuclear antigen (PCNA) immunocytochemicalstaining. The migration of HPASMCs was measured by Transwell cell test. The results showed that the eukaryoticexpression vector of antisense cyclin D1 gene was constructed and transfected into HPASMCs successfully. The cyclin D1 mRNA andprotein levels in antisense cyclin D1 group were significantly lower than those in control group (P<0.05). In 5% CSE group, the cyclinD1 mRNA and protein levels were elevated significantly compared with those in control group (P<0.05), and the indicators of cell proliferation and migration were increased as well (P<0.05). The cyclin D1 mRNA and protein levels, the indicators of cell proliferationand migration in antisense cyclin D1+5% CSE group were remarkably lower than those in 5% CSE group (P<0.05). These resultssuggest that CSE could promote HPASMCs proliferation and migration through up-regulation of cyclin D1 expression. PIRES2-EGFPascyclinD1 could attenuate CSE-induced proliferation and migration of HPASMCs by suppressing the expression of cyclin D1, whichimplicates that cyclin D1 might be involved in the process of HPASMCs proliferation and migration stimulated by CSE.

Key words: tobacco; cyclin D1; pulmonary artery; smooth muscle cells; cell proliferation; cell migration

Received: 2010-01-14　　Accepted: 2010-02-23

Corresponding author: 徐永健　　E-mail: yjxu@tjh.tjmu.edu.cn

Citing This Article：

XIANG Min, XU Yong-Jian, LIU Xian-Sheng, ZENG Da-Xiong. [Cyclin D1 is involved in human pulmonary artery smooth muscle cells proliferation and migration induced by cigarette smoke extract.] [Ariticle in Chinese] . Acta Physiol Sin 2010; 62 (2): 156-162 (in Chinese with English abstract).