[Detection of cell cycle related gene expression in cultured immortalized XXXX lens epithelial cell].

@entity1 @entity1364 @entity19 @entity15308 @entity17342 @entity1014 @entity16433 @entity994
OBJECTIVE: To investigate the gene expression of @entity1364 E, @entity15308 ) and CDK inhibitor ( @entity17342 ) @entity994 during cell cycle of the @entity1 lens epithelial cell (HLEC) for elucidating the molecular mechanism of proliferation of HLEC and the formation of @entity16433 ( @entity16433 ). METHODS: HLE-B3 cell line was cultured in vitro under MEM medium with 15% fetal bovine serum. The cell cycle was analyzed by flow cytometry (FCM) system. The cell cycle was synchronized by double @entity1014 block method. The mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). The total RNA was separated from cells at different phases. Protein expression was detected with immunohistochemistry or western blotting method. The primary antibodies were rabbit anti-human @entity1364 E polyclonal antibody, mouse anti-human @entity994 and @entity15308 monoclonal antibody, respectively. RESULTS: FCM showed that G(1) phase was about 33.5%, S phase cells about 46.1%, and the G(2) phase about @entity19 %, total cells in G(2) and S phase were over 60%. It was suggested that HLE- B3 cells have relatively more capacity of mitosis than normal LEC. The mRNA expression of @entity15308 was found at all phases of cell cycle during synchronization, but @entity1364 E mRNA only expressed during synchronized cells, and not expressed in non-synchronized cells. Only Sphase cells could express very low @entity994 mRNA. There was no any positive signal of @entity1364 E by immunohistochemistry or western blot. @entity15308 protein detected by Western blot expressed at all phases. The @entity994 protein was expressed in a few cells by immunohistochemistry, but @entity994 could not detected by western blotting. These results showed @entity1364 E and @entity15308 were higher expressed than @entity994 in transcriptional level and translational level. CONCLUSIONS: There is mRNA expression of @entity1364 E, @entity15308 and @entity994 during the cell cycle in HLE-B3 cell line, but it means the imbalance of the association of @entity17342 . These results suggest that the imbalance expression of @entity1364 , CDK and @entity994 be the molecular mechanism of @entity16433 .
