The Inhibitory Effect Of Jianpi Yangwei Decoction And Its Decomposed Formulas On BGC823/5-Fu Cell Growth

Objective1.To identificate the drug resistance of resistant cell line BGC823/5-Fu2.To observe the growth inhibitory effect of the decomposed formulas of Jianpi Yangwei Decoction on BGC823/5-Fu cells3.To observe the growth inhibitory effect of Jianpi Yangwei Decoction containing serum on BGC823/5-Fu cells4.To observe the inhibitory rate of Jianpi Yangwei Decoction and its decomposed formulas on Xenotransplantation of human gastric cancer in BGC823/5-Fu cells in nude miceMethods1.Recoverd the Human gastric cancer-resistant BGC823/5-Fu cells which were induced by low-dose shock in the early stage.The drug resistance RI was detected by MTT assay,and drug resistance was defined.2.MTT assay was used to determine the growth inhibition on the resistant cells BGC823/5-Fu and ordinary BGC823 cells by the decomposed formulas of Jianpi Yangwei Decoction in different concentrations after 48h(fructus amomi-costustoot;pericarpium citri reticulatae-rhizoma pinellinae praeparata;rhizoma sparganii-curcuma zedoary-salvia chinensis).Compared the IC50 of ordinary BGC823 cells with drug-resistant cells BGC823/5-Fu by drawing groth curve and calculating the IC50 of each decomposed formulas.3'Hoechst staining was used to observe the apoptosis of BGC823/5-Fu cells induced by the disassembled of Jianpi Yangwei Decoction.Each group were given medicine for fructus amomi-costustoot(2mg/ml;4mg/ml,6mg/ml),pericarpium citri reticulatae-rhizoma pinellinae praeparata(5mg/ml,15mg/ml,25mg/ml),rhizome sparganii-curcuma zedoary-salvia chinensis(0.25mg/ml,0.5mg/ml,lmg/ml)and control group(no drug),reaction time was 48h.4.Male Sprague-Dawley rats were given Jianpi Yangwei Decoction(2.4,1.2,0.6g/ml)by gavaging administrationg for 14 days and the blood was taken from femoral artery to prepare drug-containing serum.MTT assay was used to examine the growth inhibition of drug-resistant cells BGC823/5-Fu by different concentrations of Jianpi Yangwei Decoction containing serum,then,compared with ordinary BGC823 cells.5.By subcutaneous tumorigenesis test,observe the effects of Jianpi Yangwei Decoction and the disassembled of fructus amomi-costustoot on the suppression of tumor size in BALB/C nude mice.BALB/C nude mice were divided into control group(saline),5-Fu group(20mg/kg,intraperitoneal injection),Jianpi Yangwei Decoction group(28g/kg),fructus amomi-costustoot(2.4g/kg),Jianpi Yangwei Decoction combined with 5-Fu group(Jianpi Yangwei Decoction:28g/kg,5-Fu:20mg/kg),fructus amomi-costustoot combined with 5-Fu(fructus amomi-costustoot:2.4g/kg,5-Fu:20mg/kg)and observed the inhibitory effect of tumor size on Xenotransplantation of human gastric cancer in BGC823/5-Fu cells in nude mice by using Jianpi Yangwei Decoction,fructus amomi-costustoot or combined with the 5-Fu.Results1.The drug resistance of resuscitated drug-resistant cell BGC823/5-Fu was 10.45.2.Jianpi Yangwei Decoction and its decomposed formulas had inhibitory effects on the growth of drug-resistant BGC823/5-Fu cells and ordinary BGC823 cells after 48h.The IC50 of pericarpium citri reticulatae-rhizoma pinellinae praeparata on two cell lines respectively were(22.18±0.94)mg/ml and(21.48±0.47)mg/ml,the IC50 of fructus amomi-costustoot on two cell lines were(6.69±0.12)and mg/ml(6.46±0.15)mg/ml,the IC50 of rhizoma sparganii-curcuma zedoary-salvia chinensis on two cell lines of IC50 were(1.98±0.13)and(1.48±0.13)mg/ml.By comparison of IC50 on drug-resistant BGC823/5-Fu cells and ordinary BGC823 cells,we found that the pericarpium citri reticulatae-rhizoma pinellinae praeparata and fructus amomi-costustoot had no statistical difference(P>0.05),rhizoma sparganii-curcuma zedoary-salvia chinensis showed a statistically significant difference(P<0.05)compared with the IC50 of BGC823/5-Fu cells to BGC823 cells(P<0.05),but the difference was small.3.Apoptosis Hoechst staining experiments showed that decomposed formulas stimulated the drug resistant BGC823/5-Fu cells after 48h,the nuclei of control group were blue,the nucleus of group of amomi-costustoot(2mg/ml,4mg/ml,6mg/ml),pericarpium citri reticulatae-rhizoma pinellinae praeparata(5mg/ml,15mg/ml,drug 25mg/ml),rhizoma sparganii-curcuma zedoary-salvia chinensis(0.25mg/ml,0.5mg/ml,lmg/ml)were densely stained,or presented a chunky dense stain.With the incasing drug concentration on BGC823/5-Fu cells of pericarpium citri reticulatae-rhizoma pinellinae praeparata and amomi-costustoot,the nucleus were more densely stained.Although the group of rhizoma sparganii-curcuma zedoary-salvia chinensis may make the nucleus were densely stained,but the dense nucleus did'nt increased with the increasing of the concentration.4.The growth of BGC823/5-Fu cells and ordinary BGC823 cells were inhibited by the Jianpi Yangwei Decoction containing serum,and the inhibition rate was up-regulated with the increase of the concentration.The inhibition on BGC823/5-Fu cells and BGC823 by using middle concentration and high concentration of Jianpi Yangwei Decoction containing serum was no significant difference(P>0.05).5.Animal experimental results:Jianpi Yangwei Decoction and fiructus amomi-costustoot can inhibit the tumor size on Xenotransplantation of human gastric cancer in BGC823/5-Fu cells in nude mice,the inhibitory rate was 36.3%and 32.35%,which was higher than the group of 5-Fu(the inhibitory rate was 19.01%);Chinese medicine can reduce the tumor inhibition rate when it was used with 5-Fu(the group of Jianpi Yangwei Decoction +5-Fu:55.19%;the group of fuctus amomi-costustoot+5-Fu:48.72%)ConclusionTested again for the drug resistance of the gastric cancer-resistance cell line BGC823/5-Fu which was established in the early stage of resuscitation,the RI was 10.45.The inhibition of BGC823/5-Fu cells by pericarpium citri reticulatae-Rhizoma Pinellinae Praeparata and fructus amomi-costustoot was induced by apoptosis,and it was not related to the resistance to 5-Fu.The inhibition of BGC823/5-Fu cells by Rhizoma sparganii-curcuma zedoary-salvia chinensis was not related to the resistance to 5-Fu,and it did not cause apoptosis.We found that the inhibitory effect of Jianpi Yangwei Decoction containing serum on BGC823/5-Fu cells was rised with increasing concentrations by MTT assay,and was not related to its resistance to 5-Fu.In vivo experiments with Jianpi Yangwei Decoction and fructus amomi-costustoot showed that both can inhibit the growth of tumors,and Chinese medicine can increase the inhibition of 5-Fu with a small influence of the weight of Xenotransplantation of human gastric cancer in BGC823/5-Fu cells in nude mice.