Methotrexate combined effect of caffeine on human osteosarcoma cells in vitro observation of.doc

 PAGE \* MERGEFORMAT 7 Methotrexate combined effect of caffeine on human osteosarcoma cells in vitro observation of Abstract [Objective] methotrexate combined effect of caffeine on human osteosarcoma cells in vitro. [Method] to people osteosarcoma cell line OS-732 cells, as the research object, according to factorial experimental design reasons, set a non-drug group (negative control group), two caffeine groups, three methotrexate group, 4 caffeine, methotrexate combined treatment group (mixed drug group). Culture time, respectively MTT cytotoxicity test method and flow cytometry (FCM) analysis, observed three groups of drugs on cell toxicity (with remnants of cells indicated that the absorbance value A) and cell cycle. [Results] FCM analysis of the Group of 48 h G2 / M phase cell percentage (%) of a group (23.210 +- 0.416), 2 group (23.120 +- 0.440), 3 group (28.770 +- 0.531), 4 group (23.267 +- 0.319), 1 group and two groups no statistically significant difference, the remaining difference between the groups was statistically significant (P Will be placed in each bottle of cell culture in 48 h out to 0.25% trypsin 3 ~ 5 min, adherent cells to the sieve-like gap between the emergence date. Abandoned to the digestive juice, plus Hank’s solution. The cells from the sides of the bottle with the straw on the wind and percussion gently down and into the centrifuge tube. Short-term low-speed centrifugation (1 000 r / min, 5 min). Supernatant, add cold PBS (pH 7.4), uniform wind and percussion, and short-term low-speed centrifugation 3 times (1 000r/min, 5 min) to remove the suspension of the cell debris. Again joined the cold PBS (pH 7.4) uniform wind and percussion, made into single cell suspension. The cell sap to 500 mesh nylon mesh filter to remove overlapping into groups of cells. Adjust the cell concentration of 1 × 104/ml. At 4 ℃ refrigerator for later staining detected by flow cytometry the percentage of G2M phase cells. Each cell measured three tim