科研之友
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摘要
Background: To determine in vitro if indomethacin inhibits proliferation and induces apoptosis in human retinoblastoma cell line Y79, and to explore possibly involved signaling pathways. Methods:The human retinoblastoma cell line Y79 was cultured with indomethacin at various concentrations (0, 25, 50, 100,200 and 400 mu mol/l). The effect of indomethacin on cell proliferation and apoptosis was examined by the Cell Counting Kit-8 and TUNEL test, respectively. The mRNA level of survivin, beta-catenin and BcI-2 was detected by RTPCR. The protein level of survivin was measured by ELISA. Western blot was used to analyze beta-catenin, nuclear factor (NF)-kappa B/p65, phosphorylated Akt (pAkt) and total Akt (tAkt) expression in cultured cells. Results: Indomethacin treatment inhibits proliferation (at concentrations from 25 to 400 [mu mol/l) and induces apoptosis (at concentrations from 100 to 400 mu mol/l) of human retinoblastonna cell line Y79 in a dose-dependent manner. RT-PCR showed that the mRNA expression of BcI-2 (F = 20.497; p < 0.001) and of beta-catenin (F = 14.835; p < 0.001) was significantly different among the treated groups. Survivin mRNA levels remained steady, but its protein levels decreased significantly as measured by ELISA (F = 67.633; p < 0.001). Western blot analysis showed a dose-dependent downregulation of beta-catenin (F = 37.411; p <0.001), NF-kappa B/p65 (F = 16.302; p <0.001) and of pAkt (F = 27.700; p < 0.001) after indomethacin treatment, while tAkt protein expression was steady among the groups. Conclusions:Treatment with indomethacin can potently suppress proliferation and induce apoptosis in the retinoblastoma Y79 cell line. Wnt/beta-catenin, NF-kappa B and Akt/PKB pathways might be implicated in the process.
