| Allium sativum is the the bulb of lily - shallot category plants, it has been used as folkdrugs for several thousand years in China and other countries through out the world. The abstraction and application of all kinds of chemical ingredients from galic has been realized. Using chromatogram - mass spectrum method we can obtain five ingredients, one of which possesses stronger antifungal and antibiotic power and is called diallyl trisulfide (also named Allicin). Its structure can be determined by physics - chemical assays as CH2 = CH - CH2 - S - S - S - CH2 - CH = CH2, molecular formula is C6H10S3. It has been used not only in antibioltic and eliminating inflammation treatments, preventing infective diseases, but also widely applied in antiaging, reducing blood fat, reducing blood pressure, anticoagulant and antivirus fields. In addition to the functions mentioned above, the study of these years has found that allicin can block the synthesis of the cancerogen, inhibit the activation of the cancerogen and the effect of antimutation and antiaberration. Allicin has straight lethal effect on tumor cell; Allicin has the effect of inhibit growth of tumor cell, regulate the cell cycle, promotes tumor cell to apoptosis. Allicin can also interfere with the growth of tumor through regulating the immune function and allicin can enhance the effect of anticancer drug through the combined administration of anticancer drugs.Nowadays, chemotherapy is one of the main means of the malignant tumor's therapy. Most of the chemotherapeutics used in clinic can not only inhibit and kill the tumor cell but also damage the normal histiocyte, and then they become the obstacle of limit dose of the chemotherapy and affect the therapeutic effect greatly. So in the course of tumor chemotherapy they kill the tumor cell utmost while the development of the drug which poison is minimal to the normal cell is thought highly of. Using the cell cycle - specific drug has important significance in killing the tumor cell which is in the sensitive phase of the drug. For example , it can delay the cycle proceeding of tumor cell, prohibit the cell from one phase to the next phase, thus results in temporary accumulation of cells, the effect will be enhanced significantly at this time when be given the drug having the effect of killing, meanwhile, it can also reduce the adverse effect through reducing the dose of anticancer drugs.Allicin has the effect of regulating the cell cycle and enhance the therapy effect through the combined administration of anticancer drugs. However, there are fewer reports on combined administration of allicin. There is still much work to do in order to explore the mechanism of inhibition effects of allicin on tumor cells cycle and kill effects of it on tumor cells combined with chemotherapeutic drugs.ObjectiveTo study the effect of allicin on human gastric cells BGC - 823 and SGC -7901, to explore the combined application of allicin with chemotherapeutic drugs to these tumor cells and enhance the effects of anticancer drugs; to investigate the mechanism that allicin can reduce the tolerance of some anticancer drugs.MethodsCell cultivation and MTT method to evaluate the inhibition rates of proliferation of cells and IC50; analyze the change of cell cycle by flow cytometry ; S -P immunohistochemistry to determine the changes of expression of proteins in p21, p27, GST - it , P - glycoprotein; study the effects of combining allicin withCCSA (NVB, 5 -FU, MMC )and CCNSA(PDD) and observe the changes of cells' activities.Results1. Allicin inhibits cell growth of BGC -823 and SGC -7901 conspicuous, two kinds of gastric cells are blocked in the phase of G2/M after being handled with allicin. Allicin enhance the cell poison activity of chemotherapy drug NVB which has specific action of G2/M phase when through the combination use of it. In BGC -823 cell the IC50 of 72 hours is 30ug/ml, in SGC -7901 cell the IC50 of 72 hours is 20ug/ml . These cells are treated by allicin at the concentrations of 30ug /ml and 20ug /ml for 24 hours and 48 hours, after assays by flow cy-tometry , compared with the control group , the percentage of G0/G1 phase of the cells decreases and that of G2/M phase cells increases significantly in allicin treated group(p <0. 01). when allicin is through the combined administration of chemotherapy drug, in BGC -832 cells, the 72h IC50 value of NVB decreases from 9ug/ml to 2. 25ug/ml, the kill effect increases by three times; In SGC -7901 cells, the 72h IC30 value of NVB decreases from 43ug/ml to 12. 5ug/ml, kill effect ratio increases by 2.44 times.2. Allicin increase the expression of two kinds of gastric cells' s p21, p27 protein; After the combined administration of chemotherapy drug NVB the expression of gastric cells p21, p27 protein increase significantly. The other two kinds of cell that have not been disposed, their p21,p27 protein are both negative; While after the SGC -7901 cell has been affected by the allicin,p21 and p27 protein are both positive expression, and the expression increase with the increasing concentration of allicin, of which p21 protein' s positive expression rates are 34. 3% ,41.9% ,57.7% ,62.6% respectively after the effect of allicin whose concentrations are 5ug/ml,10ug/ml,15ug/ml,20ug/ml; p27 protein' s positive expression rates are 29. 3%,35.4% ,41.5% ,49.2% respectively after the effect of allicin whose concentrations are 5ug/ml,10ug/ml,15ug/ml,20ug/ ml; After the effect of allicin whose concentrations are 5ug/ml,10ug/ml,l5ug/ ml,20ug/ml on BGC -823 cell, p21,p27 protein' s positive expression ratesare 31.1% ,36.6% ,42. 1%,49. 8% and 25.9% 29.4% ,35.1% ,40.9% respectively. After disposing the gastric cells with chemotherapy drug and allicin, the result of p21 ,p27 protein expression are; the two kinds of cell disposed with chemotherapy drug 5 -FU(72h IC50value) ,MMC(72h IC50value) , their p2K p27 protein both express negatively, while the two kinds of cell disposed with NVB(72h IC50 value) ,their p21,p27 protein both express positively, BGC -823 cell' s p21 ,p27 positive expression rates are 48.2% and 41.4% respectively; SGC -7901 cell' s p21,p27 positive expression rates are 53.2% and 49. 7% respectively. After the two kinds of cell disposed with allicin and chemotherapy drug, the expression of p21 and p27 protein are both positive. BGC -823 cell' s NVB + allicin group,5 - FU + allicin group,MMC + allicin group , whose positive expression rates of p21 are 79.7% ,38. 2% ,40.1% respectively; positive expression rates of p27 are 66. 9% ,34.4% ,36.5%. SGC -7901 cell' s BGC -823 cell' s NVB + allicin group,5 - FU + allicin group,MMC + allicin group , whose positive expression rates of p21 are 83. 6% ,50. 1% ,51. 6% respectively; positive expression rates of p27 are 70. 8% ,30. 3% ,36.1% .3. The expression of GST - π,P - glycoprotein protein increases in two kinds of gastric cells which have been disposed by allicin while the concentration of allicin increases. It shows that allicin can decrease the expression of GST -π,P - glycoprotein. The expression of GST - π,P - glycoprotein in the two kinds of gastric cells which have not been disposed by allicin are both positive, the positive rate are: the BGC —823 cell' s GST - π protein expression is 67. 3% , P - glycoprotein expression is 48. 2% ;the SGC -7901 cell's GST - π protein expression is 59. 8% , P - glycoprotein expression is 36. 7%. After allicin has affected the SGC -7901 cell, the expression of GST — π,P - glycoprotein decreases, and the expression increases with the decreasing concentration of allicin, of which GST - π protein' s positive expression rates are 59. 8% ,48. 3% , 34.9%,25.7% ,13.6% respectively after the effect of allicin whose concentration are 0ug/ml,5ug/ml,10ug/ml,15ug/ml,20ug/ml; P - glycoprotein' s positive expression rates are 36. 7% ,34. 3% ,21. 4% ,13. 5% ,9. 2% respectively after the effect of allicin whose concentration are 0/ml, 5ug/ml,10ug/ml,15ug/ ml,20ug/ml; After the effect of allicin whose concentration are 5ug/ml,10ug/...
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