Effect Of Phlegm Sanjie On Drug Resistance Cell Lines Of Human Breast Cancer And The Mechanisms

Objective:Study the effect of phlegm Sanjie on resistance-associated protein and apoptosis gene. Elucidate the multidrug resistance reversal mechanism. Provide theoretical and experimental basis for the problem of multidrug resistance.Methods:1. The effect of Phlegm Sanjie on MCF-7/ADR cell multidrug resistance -associated proteins and genes1) To detect resistance reversal effect of Phlegm Sanjie on MCF-7/ADR cell by MTT assay;2) To detect the expression of P-gp on MCF-7/ADR cell by flow cytometry after the serum containing was used;3) To detect the expression of MRP on MCF-7/ADR cell by Western Blot;4) To detect the expression of MDR1 and MRP1 mRNA in MCF-7/ADR cell by RT-PCR assay;2. The proliferation and induction of apoptosis of the Phlegm Sanjie on MCF-7/ADR cell1) By cloning experiment, we observed the effect of Phlegm Sanjie on MCF-7/ADR cell colony formation and calculated cloning efficiency;2) To detect the effect of Phlegm Sanjie on cell cycle by the flow cytometry; 3) After the serum containing was used, we observed the morphological changes by fluorescence microscopy and electron microscopy.4) To detect the effect of Phlegm Sanjie on apoptotic gene Bcl-2 by the flow cytometry;Result:1. The effect of Phlegm Sanjie on MCF-7/ADR cell multidrug resistance -associated proteins and genes1) Phlegm Sanjie serum containing could significantly inhibit the proliferation of MCF-7/ADR cells. And with the increasing of serum containing concentrations and action time, the inhibition enhanced.2) Serum containing could significantly inhibit the expression of P-gp. Compared with the control group, there was statistical significance (P<0.01). Containing serum could enhance the effect of ADM in apoptosis induction.3) ADM could increase the expression of MRP.Serum containing could decrease the expression of MRP.4) MDRl and MRP1 mRNA expressed highly in MCF-7/ADR cells. After different concentrations of serum acted on the MCF-7/ADR cell, the expression of MDRl and MRP1 mRNA decreased. And it showed a concentration-dependent.2. The proliferation and induction of apoptosis of the Phlegm Sanjie on MCF-7/ADR cell1) With the concentration increasing, the cell colony formation rates decreased gradually.2) After serum containing was used, the resistant cell lines were arrested at G0-G1 phase.The proliferation of resistant cell lines were prevented by activating apoptosis. The induction of apoptosis existed dose-effect relation ship.3) Microscopic observation showed that with the increasing of serum containing concentration and action time, the proportion of apoptotic cells increased.4) Serum containing group could significantly down the expression of apoptosis gene Bcl-2. And it showed a dose-dependent (P<0.05). Conclusion:1. Phlegm Sanjie serum containing could act on the resistance-associated proteins and genes of MCF-7/ADR cell. The mechanism may be related to reducing the expression of MDR1and MRP, and increasing the drug concentration in MCF-7/ADR cell.2. Phlegm Sanjie could inhibit tumor proliferation and induce apoptosis.The mechanism may be acting on G1/S node, so that resistant cell lines were arrested at G0-G1 phase. The other mechanism may be decreasing the expression of Bcl-2 and preventing the proliferation of resistant cell lines by activating apoptosis.