Effect of Cyclin A on origin recognition complex-1 expression of rat vascular smooth cells
10.3760/cma.j.issn.1008-6315.2014.09.001
- VernacularTitle:细胞周期蛋白A对培养大鼠血管平滑肌细胞起始识别复合物1表达水平的影响
- Author:
Minghong JIANG
;
Maoqin SHU
- Publication Type:Journal Article
- Keywords:
Cyclin A;
Vascular smooth muscle cells;
Origin recognition complex 1
- From:
Clinical Medicine of China
2014;30(9):897-901
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of Cyclin A on origin recognition complex-1 (ORC1) expression of rat vascular smooth cells (VSMCs).Methods Primary VSMCs of thoracic aorta in rats was obtained by the adherence method of tissue culture.The synchrony of cell was obtained by the method of double-thymidine block.In different cell cycles of VSMCs,the expression of ORC1 mRNA was determined by RT-PCR and the protein expression of ORC1 was observed by flow cytometry.Results Synchronized VSMCs were obtained and identified by the methods of double-thymidine block,colchicine treatment and serum starvation.Synchronized growth was monitored by flow cytometry.All the synchronized VSMC's distribution ration was (89.22±3.54) ％ at G0/G1 phase,(66.74 ±7.16)％ at G1/S phase,(63.24 ±4.06)％ at S phase and (51.64 ± 11.18)％ at G2/M phase and there was statistically significant difference compared with other phase(P ＜0.01).There was no significant effect of Cyclin A on ORC1 mRNA expression at a quiescent stage of VSMC.At G2/M phase peaked ORC1 was (52.133 3 ± 2.122 1)％,at G1/S phase was(10.916 7 ± 0.531 1)％,at S phase was (7.656 7 ± 0.412 4)％,and there was statistically significant difference (P ＜0.01).At G2/M phase ORC1 downed to the lowest point was (1.276 7 ± 0.161 7) ％,at G1/S phase was (13.371 0 ± 1.057 3)％,at S phase was (3.043 3 ± 0.538 0)％,and there was statistically significant difference (P ＜ 0.01),suggesting that Cyclin A might prevent ORC1 binding the chromatin of VSMCs.Conclusion Cyclin A may be an important regulative factor at the initiation of ORC1 in VSMCs.
