title: [The effect of cigarette smoke extract on the proliferation of human airway smooth muscle cells sensitized by serum from bronchial asthmatic patients].
abstract: OBJECTIVE
  To study the effect of cigarette smoke extract (CSE) on the proliferation of human airway smooth muscle cells (HASMCs) sensitized by serum from asthmatic patients and the underlying mechanisms.
  
  
  METHODS
  HASMCs were cultured from primary generation. Cells between passage 4 and 8 were used in the study. HASMCs were sensitized by 10% serum from asthmatic patients and were divided into an asthmatic serum group, an asthmatic serum + CSE group, an asthmatic serum + GW8510 (inhibitor of cyclin-dependent kinase-4) group and an asthmatic serum + CSE + GW8510 group. Non-asthmatic human serum treated HASMCs served as the control. The proliferation of HASMCs was examined by cell cycle analysis, MTT colorimetric assay and [(3)H] thymidine incorporation. The expression of cyclinD(1) was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting.
  
  
  RESULTS
  The percentage of S + G(2)/M phase, the absorbance (A) value and the DNA synthesis value in asthmatic serum group were significantly increased compared with those of the control group (q = 6.25, 5.61, 6.82, respectively, all P < 0.01). The percentage of S + G(2)/M phase, the absorbance (A) value and DNA synthesis value in the asthmatic serum group were (21.4 ± 1.1)%, 0.392 ± 0.124 and 2669 ± 138, respectively. Their value in the asthmatic serum + CSE group were (33.3 ± 1.3)%, 0.612 ± 0.201 and 3552 ± 303, respectively, which were significantly increased compared with those of the asthmatic serum group (q = 5.67, 6.32, 5.56, respectively, all P < 0.01). Their value in the asthmatic serum + GW8510 group were (14.7 ± 1.4)%, 0.301 ± 0.097 and 1812 ± 109, respectively, which were significantly decreased compared with those of the asthmatic serum group (q = 6.02, 5.53, 5.79, respectively, all P < 0.01). The ratios of A value of cyclinD(1) mRNA and the expression of cyclinD(1) protein in the asthmatic serum group were 0.291 ± 0.112 and 0.186 ± 0.002, respectively. The ratios of A value in the asthmatic serum + CSE group were 0.521 ± 0.102 and 0.312 ± 0.002, respectively, which were significantly increased compared with those of the asthmatic serum group (q = 12.09, 9.26, respectively, all P < 0.01). The ratios of A value in the asthmatic serum + GW8510 group were 0.223 ± 0.038 and 0.150 ± 0.002, respectively, which were significantly decreased compared with those of the asthmatic serum group (q = 6.86, 5.60, respectively, all P < 0.01).
  
  
  CONCLUSIONS
  HASMCs sensitized by serum from asthmatic patients showed accelerated proliferation after intervention by CSE, with increased expression of cyclinD(1). CSE may increase the proliferation of HASMCs sensitized by serum from asthmatic patients via regulating cyclinD expression.
authors.affiliation: Department of Respiratory Medicine, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: Xiao-yu
authors.initials: XY
authors.lastname: Zhang
authors.forename: Li-jun
authors.initials: LJ
authors.lastname: Ma
authors.forename: Yong-jian
authors.initials: YJ
authors.lastname: Xu
authors.forename: Xian-sheng
authors.initials: XS
authors.lastname: Liu
authors.forename: Zhen-xiang
authors.initials: ZX
authors.lastname: Zhang
mesh_terms: D000328:Adult; D001249:Asthma / Q000097:blood / Q000378:metabolism* / Q000473:pathology*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D019938:Cyclin D1 / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D012137:Respiratory System; D044967:Serum / Q000737:chemistry; D015398:Signal Transduction; D012906:Smoke / Q000009:adverse effects*; D014026:Nicotiana / Q000009:adverse effects*; D055815:Young Adult
chemical_list: C530104:CCND1 protein, human; D012906:Smoke; D019938:Cyclin D1