Investigate The Effects Of Human Gastric Cancer Cell SGC-7901 By Oridonin Based On Cell Cycle Synchronization

The Oridonin injection from Labiatae Isodon is a a separation plant Cardamine Sam kaurene diterpenoid natural compounds.Oridonin injection has a unique and significant anti-tumor activity,Significantly inhibit or kill a variety of tumor cells in breast cancer,prostate cancer,gastrointestinal cancer and blood system tumors and its toxicity is low,and no significant damage to important organs of the body such as the liver,kidney,bone marrow,and other.The oridonin injection research on gastric cancer cells less in the country,in this research will study the effects of human gastric cancer cell SGC-7901 by oridonin based on cell cycle synchronization.This research group have been studied the effects of human gastric cancer cell SGC-7901 by oridonin.In this study we will use the vitro experiments to study the effects of human gastric cancer cell SGC-7901 by oridonin based on cell cycle synchronization and preliminary identification of the specific organization period.Using growth curve combine with formula derivation determine human gastric cancer cell line SGC-7901 cell cycle length;BrdU-FITC/PI double staining,flow cytometry determine the proportion of cells in S phase and speculate S phase length;PI staining,flow cytometry determine the effect of synchronization result by TdR double-blocking synchronization method;PI staining,flow cytometry determine the effects of human gastric cancer cell SGC-7901 by oridonin based on cell cycle synchronization;Western Blotting analysis oridonin Cyclin B1 and Cyclin A to determine oridonin arrest of human gastric cancer SGC-7901 cells specific period.The experimental results show,(1)counting method recorded separately the total number of human gastric cancer SGC-7901 cells at 0h,24h,48h,72h,96h,120h,144h and 168h,making logarithmic growth curve and fitting growth cuirvc by Excel,Tc=?t X 24h X Lg2/(LgNt-LgN0)]derivation to Tc=24h/b,calculated the human gastric cancer SGC-7901 cells logarithmic phase cell cycle length is Tc=30.63h;(2)the result of BrdU-FITC/PI double staining,flow cytometry determine the proportion of cells in S phase and speculate S phase length show that,Adding BrdU for 1h,3h,5h and 7h.the percentage of S phase are 18.5%,20.9%,24.6%,27.8%,calculate the length of S phase is 10.53h;(3)the result of PI staining,flow cytometry determine the effect of synchronization result by TdR double-blocking synchronization method show that,the cell group adding TdR with final concentration of 2mmol/L have been synchronized in G0/G1 phase,TdR double-blocking method can indicate the human gastric cancer SGC-7901 cells for cell cycle synchronization,establish synchronization model;(4)the result of PI staining,flow cytometry determine the effects of human gastric cancer cell SGC-7901 by oridonin based on cell cycle synchronization show that,after cell cycle synchronization,adding a final concentration of l,2,4,8,16,32?mol/L of oridonin,24h group compared with the control group,the proportion of cells in S phase gradually increased,percentages are 28.6%,27.7%,52.6%,60.0%,59.7%;48h group compared with the control group,the proportion of cells in G2/M phase gradually increased,percentages are 41.5%?46.5%?47.6%?53.7%?56.9%?63.4%,compared to the control group is statistically significant.Indicate that oridonin can have an impact on human gastric cancer SGC-7901 cell cycle,in 48h after administration of the cells arrested in the G2/M phase;(5)the result of Western Blotting analysis oridonin Cyclin B1 and Cyclin A to determine orridonin arrest of human gastric cancer SGC-7901 cells specific period show that,oridonin can affect the expression of Cyclin A and Cyclin B1,compared with the normal group and control group cells,the expressiort of Cyclin B1 didn't chang,and Cyclin A is basically not expressed,we concluded Oridonin specific period of human gastric cancer SGC-7901 cells for M phase arrest.Through the above experiments,after the establishment of the cell cycle synchronization model clearer found that oridonin on human gastric cancer SGC-7901 cell cycle through their arrest in G2/M phase,and specifically period arrest M phase.