Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences · 2009
[Construction of adenoviral vector encoding Calponin-1 siRNA and its effect on human myometrium cells in vitro].
Аннотация на английскомСтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
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Gu Yonghong, Zhou Changju, Hu Lingyu, Chen Qian, Zhang Weishe
Аннотация
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. Adenovirus 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.
METHODS: Human uterine smooth muscle tissues were digested with enzymes, cultured and confirmed with immunocytochemistry. Adenovirus 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.