| Objective:To ananlyze the inhibitory action of paclitaxel liposome to breast cancer cell (MCF-7) and ovarian cancer cell (SKOV-3), observe the time-concentration effect of this drug and explore the mechanism of paclitaxel liposome effecting on the tumor cell in order to provide basal confirm for clinical application on breast and ovarian cancer.Methods:MCF-7 and SKOV-3 cells were cultured and divided groups, added with different concentration of paclitaxel liposome and observed the time-concentration effect of the drug by MTT. The application of flow cytometry analysed the inhibitory rate of cell and the cell cycle of each group and the expressions of bcl-2 and bax proteins were detected by western-blot to explore the mechanism of the drug effects on above two cell lines.Results:1. The best planting density of the cell lines MCF-7 and SKOV-3 for the 96 well plate was 4×104/ml.2. When the planting density was identical, MCF-7 cells grew faster than SKOV-3 cells, and the growth tendency of the two cell lines coincided with the S growth curve.3. Used MTT colorimetric to screen the effective concentration and time of paclitaxel liposome, which demonstrated that the drug could inhibit MCF-7 cells and SKOV-3 cells and the inhibition rate showed time-concentration-dependent effect. Under the same concentration of paclitaxel liposome, the inhibition rate increased along with the time extension; while under the same reaction time, the inhibition rate increased along with the increase of drug concentration. Three concentrations(1μg/ml,5μg/ml,20μg/ml) and two reaction time 24h and 48h were chosen.4. Normal MCF-7 and SKOV-3 cells possessed the characteristics:polygon shape, adherence growth, high breeding speed; had integrated cell membrane and limpid borderline. After be treated with paclitaxel liposome, along with the increasing concentration and prolonged time, cell morphology appeared several changes:cell borderline became round, vacuole generated in cytoplasm, cell membrane disrupted, cells were solved and dead. 5. The analysis of cell cycle with flow cytometry presentated that paclitaxel and paclitaxel liposome could both block MCF-7and SKOV-3 cells at G2/M phase, G0/G1 phase and S phase were obviously decreased after treated with drugs. The cell cycle blocking between two groups (Taxinol and Taxinol liposome) had no significant difference. The changing of drug concentration and reaction time could not influence cell cycle blocking and the blocking effect could improved along with increasing drug concentration and prolonged reaction time.6. The analysis of apoptosis with flow cytometry presentated that cell apoptosis was more obviously along with increasing drug concentration. After statistical analyses by using SPSS 16.0 software, we found that under the same drug concentration, the rate of inhibition increased along with the prolonged reaction time, and there was significant difference between 24h and 48h groups. Under the same reaction time, there was no significant difference during different drug concentration groups. Meanwhile, there was no significant difference about cell blocking between Taxinol and Taxinol liposome groups.7. The western blots results demonstrated that after using paclitaxel liposome to deal with the MCF-7 and SKOV-3 cells, the expression of bcl-2 protein obviously decreased, while the expression of bax protein increased and the value of bax/bal-2 up-regulated.Conclusions:1. Paclitaxel liposome had inhibitory action and time-concentration-dependent effect on breast (MCF-7) and ovarian (SKOV-3) cancer cell line as same as paclitaxel. Therefore,paclitaxel liposome has a good future for patients with breast or ovarian cancer.2. The cell cycle blocking of paclitaxel liposome group was similar with paclitaxel group, and both groups were at G2/M phase.3. Paclitaxel liposome could induce the down-regulation of bcl-2 protein expression and the up-regulation of bax protein expression, which could be one of the drug mechanisms of action.
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