| Objective: To investigate the regulatory effect of OVCA1 on proliferation and expression of cyclinD1,p16,NF-κB in human ovarian carcinoma cell line A2780 in vitro.Method: The ovarian carcinoma cell line A2780 (A2780) and OVCA1 stably expressing A2780 were cultured in vitro. The effect of OVCA1 on proliferation in A2780 was detected by colony formatting cell culture test. The regulatory effct of OVCA1 on cyclinD1,p16 expression at gene level was detected by reverse transcription-polymerase chain reaction (RT-PCR). The regulatory effect of OVCA1 on cyclinD1,p16,NF-κB expression at protein level was detected by immunocytochemistry (ICC). Meanwhile, the regulatory effect of OVCA1 on cyclinD1,p16 expressotion at protein level was also detected by immunofluorescence (IMF).Result: 1. After cultured 3 days, we can see the liferation rate of A2780-OVCA1 is lower than A2780. The colony formatting percent of A2780-OVCA1 was obviously lower than that of A2780 (P<0.01). 2. Analysis by RT-PCR on the expression of intracellular cyclinD1 mRNA and p16 mRNA show the expression of cyclinD1 mRNA was down regulated (P<0.01), and the expression of p16 mRNA was up regulated (P<0.01). 3. Analysis by ICC and/or IMF on the expression of intracellular cyclinD1 protein and p16 protein show the expression of cyclinD1 protein was down regulated (P<0.01), and the expression of p16 protein was up regulated (P<0.01). 4. there was no correlation between the expression of cyclinD1 protein/mRNA and p16 protein/mRNA (P>0.05). 5.The inhibition of cyclinD1 by OVCA1 in A2780 cell at protein and mRNA level has no difference(P>0.05); That of p16 on protein level is 38.9152±2.8227%,which is lower than 91.3944±2.4942% on mRNA level (P<0.01). 6. The expression of NF-κB protein in cell cytoplasm and nucleus had no... |
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