Experimental Study On The Effect Of Qingdu Suppository On Cancer Of The Cervix Prevention And Cure

Objact :To investigate the effect of the Serum pharmacological effect Qingdu Suppository on the SiHa cell growth in vitro.To approach the mechanism of action on Cell biology and Molecular biology level which Qingdu Suppository inhibiting the SiHa cell growth.Method:1. the Serum Containing Qingdu Suppository preparation :In accordance with the methode of Liyikui's to extract the Serum Containing Qingdu Suppository and Blank Serum , Baofukang Suppository , Inf-αcontrolling group . Inactivate the Serum by waterbath at 56℃for half an hour. Store in -70℃Refrigerator. The Serologic pharmacological method.2. the SiHa cell growth in vitro:After different density and different Serum treating to SiHa cells 72h , we evaluated photoabsorption value ,calculating inhibition ration and getting quantity and effect data .After treating SiHa cells by Serum 24,48,72,96h, we evaluated photoabsorption value ,calculating inhibition ration and getting quantity and effect data .3. To detect the effect of cell cycle and apoptosis to SiHa cells treated with Serums by Flow cytometry After 8% density Serum treating SiHa cells 72h ,fixing cells ,digeting half an hour by RNAase ,dyeing 1h by PI away from light ,we detected it , anayzeed cell cycle by Modi Fit Sofeware and draw a DNA profile . 4. To detect the effect of regulating to p53-mRNA and MDM2-mRNA expression of SiHa cells te reated with Serums by RT-PCR Semiquantitative detection methode:After 8% density Serums treating SiHa cells , we extracted RNA , did RT-PCR reaction ,collected image by electrophoresis and gel image analytical apparatus and anayaed gray scale.Results:1. The Suppress rate of cells treated with Qingdu Suppository Serum were Significantly higher in 8% -72h group than those in the control group treated with Rat Serum,and those treated with IFN Serum were also significantly higher than those in the control group treated with Rat Serum .2. Cell cycle and apoptosis : 2.1 Cell ratio had Significantly higher at G0-G1 phase by treating with 8% -72hr OR-group Qingdu Suppository Serum ,on the other hand that Cell ratio had Significantly lower at S phase by treating with 8% -72hr group Qingdu Suppository Serum. But there was no obviously change at each phase by treating with 8% -72h V-group Qingdu Suppository Serum.2.2 The 8% -72h OR-group Qingdu Suppository Serum accelerated SiHa cell apoptosis,and so did 8% -72hr OR-group IFN Serum . But there no apoptosis peak appear in V-group Serums.3. p53 and MDM2 mRNA expression:3.1 The suppress rate of MDM2 gene expression of cells treated with were Significantly lower in than those in the control group treated with Rat Serum,and those treated with IFN Serum were also Significantly lower than those in the control group treated with Blank Rat Serum .3.2 The Suppress rate of p53 gene expression of cells treated with Qingdu Suppository Serum were significantly higher in 8% -72h group than those in the control group treated with Rat Serum ,and those treated with IFN Serum were also significantly lower than those in the control group treated with Rat Serum .ConcluSion: The Qingdu Suppository Serum suppress tumor by direct repression the cervical cancer cell proliferation , regulate cell cycle, accelerated SiHa cell apoptosis and also regulate the MDM2 - p53 feedback loop molecule mechanism.