Studies On Paclitaxel On Canine Mammary Tumors Chmp And Chmm Cell Cycle Regulatory Factors

Canine mammary tumors is a neoplastic diseases in clinical veterinary. Canine breast cancer has many likenesses with human breast cancer, as an excellent model for human breast cancer. The research showed that there is a large proportion of malignant tumors which invasion to other organs in clinical diagnosed. Tumor occurrence, development and transfer are related to the abnormal regulation of cell cycle network closely. The study in-depth of it will open up a new idea to tumor control and treatment. It can provide a new role of target not only for tumor immunology therapy and anti-cancer drug development but also for the various key points which are in the cell cycle regulatory network to control cell growth, division to prevent the proliferation of cancer cells. Taxol was developed in the late20th century, it is an anti-cancer natural drug, and it has been used in the clinical treatment of tumors widely. The mechanism of paclitaxel in the primary and metastatic tumors has not yet been report. In this experiment, a specific series of studies to solve this problem has been done.The natural occurring canine mammary tumors in primary and metastasis cell lines vitro CHMp and vitro CHMm are collected, by the concentration of Oμmol/L,0.1μmol/L,1μmol/L,10μmol/L paclitaxel for Oh,12h,24h,48h,72h respectively. Using the LightCycler2.0real-time quantitative PCR instrument and SYBR dyemethod method to test CyclinDl mRNA, p27kip mRNA, PCNA mRNA, bcl-2mRNA, p53mRNA in vitro CHMp and vitro CHMm. Used the LightCycler Software4.1to analysis the experiment data, plotted the standard curve, obtained the amplification efficiency of the reference and target gene, calculated the control group CT, blank group CT and the experimental group CT.The result showed that:paclitaxel can induce the primary and metastic canine mammary tumors vitro CHMp and CHMm apoptosis. In CHMp, CyclinDl mRNA was lowered, significant different after24h (P<0.01), but in CHMm, significant different after1μmol/L paclitaxel12h (P<0.01).In CHMp, p27kip mRNA was increased, significant different in1μmol/L paclitaxel for12h (P<0.05), and for48h p27kip mRNA was increased to the highest. In CHMm, significant different after12h (P<0.01), and0.1μmol/Lpaclitaxel for72hours was increased to the highest.In CHMp and CHMm, PCNA mRNA was lowered by the concentration and time of paclitaxel, significant different among each group (P<0.01), it was the concentration-time effect. In CHMp, bcl-2mRNA was lowered, significant different after24h (P<0.05), and paclitaxel can make it to the lowerest. But in CHMm, bcl-2mRNA lowed significantly when1μmol/L paclitaxel for12-24h.In CHMp, p53mRNA was increased when0.1μmol/L paclitaxel for48h, and difference was significant (P<0.01). In CHMm,1μmol/L24h was more than0.01μmol/L48h, and the difference was significant (P<0.01).