The Inhibitory Effects Of Podocarpus Seeds And Its Tray Extract On Human Gastric Cancer Cells

Objective: The purpose of this paper is to use in vitro experiment with drugs, preliminary screening has inhibitory effect on tumor cells of podocarpus seeds extracted parts, makes a tentative discussion of podocarpus seeds extract on the antitumor mechanisms according to flow cytometry, real-time quantitative Polymerase chain reacion technology.Each year about six hundred and forty thousand people died of stomach cancer. At present, the treatment of gastric cancer with surgery is given priority to, such auxiliary therapy as chemotherapy, radiation therapy and molecular target therapy. It is worth mentioning that traditional Chinese medicine as a cultural treasure of our country, with the tiny side effects, can relieve pain due to disease or radiation and chemotherapy patients to improve the postoperative survival rate.At the same time, traditional Chinese medicine plays an important role in the treatment of gastric cancer.Podocarpus macrophyllus(Thunb.)D.Don belongs to a kind of pine trees, having many other names. Some researches show that Podocarpus plants of containing the composition such as biflavoneand apigenin Have certain prevention and cure function on gastric cancer, liver cancer, breast cancer, ovarian cancer and other cancers. Because there is few podocarpus seeds extracts in vitro antitumor research, we use human gastric cancer cells MGC-803 as the research object to have a Preliminary discussion podocarpus seeds extract on inhibition of tumor cells, as to provide theoretical basis for further study on its antitumor mechanism.Methods: 1. The preparation of podocarpus seeds extract: separation the podocarpus seeds and its tray, crushing them, then respectively use water, 95%, 75%, 50% ethanol extraction, get the different part of podocarpus solid extracts, to freeze drying of extract, set aside. 2. Screening the active sites of Podocarpus seeds and its tray extract : based on the in vitro cultured human gastric cancer cell MGC-803 as the research object, chooses MTT method to measure the value of podocarpus seeds and its tray extract for MGC-803 cells inhibition rate, half the mortality rate as evaluation index selected podocarpus seeds and its tray extract active site. 3. To determine the best drug concentration: put the selected podocarpus seeds and its tray extract active site configuration into the solution of different concentration gradient, through the MTT test, looking for the best dosing active site concentration. 4. Using flow cytometry studies podocarpus seeds and its tray extract active site of the influence of the cell cycle of MGC-803 gastric cancer cells. 5. Using real-time quantitative Polymerase chain reacion technology researches the active site of the podocarpus seeds and its tray extract on human gastric cancer cell MGC- 803, the relativeexpression of the related genes in tumor cells the Bc L- 2, P53, C-myc and Bax.Results: 1. Determined by MTT screening experiment results show that Podocarpus seed extract for MGC- 803 gastric cancer cells having growth inhibition, the active site is 95% ethanol extract, when the strength of solution reaches 50μg/ml, affect MGC- 803 cells for 48 hours, the inhibition of human gastric cancer cell rate is 68.38%, and the relationship between strength and action time were positively correlated. 2. The flow cytometry instrument detection of MGC- 803 gastric cancer cell apoptosis rate of the experimental results show that the MGC- 803 cells apoptosis rate and podocarpus seed extracts dosage of 95% ethanol concentration was positively related to relationship, with the increase of drug concentration, cell apoptosis rate has been increased. 3. The results of real-time quantitative Polymerase chain reacion experiment show that compared with the control group, different podocarpus seed extract of 95% ethanol concentration parts on the MGC- 803 cells, significantly lowered the apoptosis related gene Bc L- 2 and C- myc, raising the gene P53 and Bax, In other words, the m RNA level of Bc L-2 and C-myc along with the increase of drug concentration significantly decreased(p < 0.05), the m RNA level of P53 and Bax is significantly enhanced with the increase of drug concentration(p < 0.05).Conclusion: Podocarpus seed extract on human gastric cancer cells in vitro MGC- 803 has obvious inhibition of value-added, the active site is podocarpus seed 95% ethanol extraction, and with the increase of the extract concentration and function of the extensionof time, the inhibition rate of gastric cancer cell is on the rise, this may be related to it can regulate the expression of apoptosis genes.