647 Investigation of Role of Different PI3K Isoforms in Breast Cancer

Abstract

division are linked with circadian system. In addition core clock factors not only participate in transcription regulation of cell cycle related genes, but also can interact directly with the cell cycle pathway. Our aim was to characterize gene expression pattern in mice Leydig tumor cell lines: MLTC-1, I-10 compared to normal mouse Leydig cells TM3, and in mice testes, vas deferens, epididymis and prostate. Quantitative Real TimePCR (qPCR) was assessed to determine Cyp19, Per1, Bmal, Clock, Cry1, Cry2 mRNAlevels. Material and Methods: All cells were cultured inthe medium under conditions recommended by manufacturer ATCC. To induce and synchronize molecular oscillator in cultured cells standard growth medium was exchanged with serumrich medium (50%). After 2 hr serum rich medium was replaced with serumfree cell adequate medium. Male C57B1/6J mice aged 10−15 weeks were fed ad libitum and maintained at 12:12 LDc ycle. Total RNA from cultured cells and mice tissues was extracted byTriReagent and converted to cDNA. qPCR was assessed to determine mRNA levels. Results: Our results show rhythmic expressionof core clock genes in mice reproductive tract, as well as in Leydig cultured cells. Moreover expression manner of those genes differs in immortalized and tumor cell lines. In all of studied parts of male reproductive tract of mice the expression of Cyp19 gene is rhythmic. Although expression pattern differs among investigated tissues. Cyp19 also shows rhythmic expression in Leydig cell lines, and consequently tumor cells exhibit altered expression profile compared to normal cells. Conclusions: Our data may provide new evidence that the core molecular oscillator and aromatase Cyp19 gene are linked to mechanism of cancer development.