Construction of adenoviral vector encoding Calponin-1 siRNA and its effect on human myometrium cells in vitro
- VernacularTitle:调宁蛋白基因siRNA重组腺病毒载体构建及其对体外子宫平滑肌细胞的作用
- Author:
Yonghong GU
;
Changju ZHOU
;
Lingyu HU
;
Qian CHEN
;
Weishe ZHANG
- Publication Type:Journal Article
- Keywords:
myometrium cells;
Calponin-1;
adenovirus vector;
delivery start
- From:
Journal of Central South University(Medical Sciences)
2009;34(12):1231-1237
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of Calponin-1 suppression on human myometrium cells through adenovirus mediated siRNA. Methods Human uterine smooth muscle tissues were digested with enzymes, cultured and confirmed with immunocytochemistry. Aadenovirus siRNA-Calponin-1 plasmid was transfected into primary cultured uterine smooth muscle cells in vitro. The expressions of Calponin-1 mRNA and protein were analyzed by RT-PCR and Western blot, respectively.Results The pAdEasy-pShuttle-U6-Calponin-1 siRNA plasmid was successfully constructed, and Calponin-1 siRNA mediated by recombinant adenovirus resulted in markedly reduced expression of Calponin-1 mRNA and protein in human myometrium cells. The gray values of Calponin-1 mRNA in the uterine smooth muscle cells in the experimental, blank control, and empty vector groups were 316.3±39.2, 1048.5±126.4 and 1027.2±127.5, respectively. The gray values of Calponin-1 protein were 323.3±43.2, 1021.5±143.4, and 1019.2±144.5,respectively. The difference between the experimental group and the blank control group as well as the empty vector group was significant (P< 0.05). There was no significant difference between the empty vector group and the blank control group (P>0.05).Conclusion The pAdEasy-pShuttle-U6-Calponin-1 siRNA plasmid can inhibit the expression of Calponin-1 in human myometrium cells in vitro,which may be a useful approach to determine the role of Calponin-1 in delivery.
