| OBJECTIVE:To investigate the effect of proto-oncogene c-erbB2 on the initiation of primordial follicles and its role in mediating the important primordial follicles'promotor EGF in this process by using RNA interference.METHODS:.Culture neonatal rat ovary in vitro and transfect the ovary with siRNA targeting proto-oncogene c-erbB2 mediated by metafactene then measure the expression of c-erbB2 by RT-PCR,and western blot in-synchronism. observe the development of primordial follicles.RESULTS:1. RT-PCR showed that c-erbB2mRNA were expressed in rat ovary. The sequence of the product amplyfied by RT-PCR is matched 100% with the sequence of c-erbB2 in Gnebank.2. After siRNA transfecting, RT-PCR showed that the levels of c-erbB2 mRNA expression were reduced significantly(,P<0.05). The inhibition rate is 49.6%,46.7% and 82.6% respectively. Meanwhire western blot showed C-erbB2 protein expression also reduced(P<0.05).3. Choose the one siRNA of the most effective then transfect it into cultured ovaries,After 8 days′culture ,the result showed that most primordial follicles have been initiated to be primary follicles in control group ;Compared to control group, ovaries treated with siRNA, the numbers of primary and second follicles were decreased significantly (P<0.01);The group treated with EGF which numbers of primordial follicle decreased while the second follicles increased significantly (P<0.01)There is no difference between control group and group treated with EGF +siRNA.CONCLUSIONS:1. The expression of proto-oncogene c-erbB2 are presented in rat ovary 2. c-erbB2 si RNA can supress the expression of the target gene c-erbB2 specifically.3.The development of primordial follicles were supressed as the expression of proto-oncogene c-erbB2 was supressed .EGF can promote the development of primordial follicles while siRNA can block this action by supressing the expression of c-erbB2 .
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