Heat shock protein 10 regulated apoptosis of mouse ovarian granulosa cells.
OBJECTIVE
To study the roles of heat shock proteins10 (HSP10) in the regulation of mouse ovarian granulose cell (GC) apoptosis, and to further define the possible roles of HSP10 in the development of polycystic ovary syndrome (PCOS).


METHODS
Mouse HSP10 small interfering RNA (siRNA) and recombinant adenoviruses overexpressing HSP10 were constructed and subsequently transfected into cultured mouse ovarian GCs. After an infection period of 48 h, the expression levels of the HSP10 gene in mouse GCs were confirmed by Western blot. The GCs were also assessed for apoptosis using flow cytometry and the TUNEL assay. Apoptosis of GCs overexpressing HSP10 was assessed by flow cytometry after cisplatin treatment.


RESULTS
Compared with control group, the expression of HSP10 was decreased in mouse GCs infected with AdCMV-siRNA/HSP10, whereas mouse GCs infected with AdCMV-HSP10 showed increased HSP10 expression p < 0.05. Knock-down of HSP10 in mouse GCs significantly increased apoptosis (p < 0.05), whereas overexpression of HSP10 significantly suppressed apoptosis induced by cisplatin (p < 0.05).


CONCLUSIONS
In the present primary study, we have successfully employed recombinant adenovirus technologies to modulate HSP10 gene expression in mouse GCs, and examined the effects on apoptosis. Our experiments have demonstrated that knock-down of HSP10 induces apoptosis of mouse ovarian GCs, whereas overexpression of HSP10 suppresses apoptosis. These findings suggested that HSP10 may play a role in the regulation of apoptosis of mouse ovarian GCs.
Ling
Jing
J
Center of Clinical Reproductive Medicine, First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Zhao
Kaokao
K
Cui
Yu-Gui
YG
Li
Ying
Y
Wang
Xinru
X
Li
Mei
M
Xue
Kai
K
Ma
Xiang
X
Liu
Jia-Yin
JY
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D015153:Blotting, Western; D018835:Chaperonin 10 / Q000235:genetics / Q000502:physiology*; D002945:Cisplatin / Q000494:pharmacology; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression; D020868:Gene Silencing; D006107:Granulosa Cells / Q000502:physiology*; D020287:In Situ Nick-End Labeling; D051379:Mice; D011085:Polycystic Ovary Syndrome / Q000209:etiology; D034741:RNA, Small Interfering / Q000235:genetics; D011994:Recombinant Proteins; D014162:Transfection
D018835:Chaperonin 10; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D002945:Cisplatin