The Effect Of 5-fluorouracil Combined With Triptolide On Pancreatic Cancer In Vitro

Objective To examin the effects of triptolide (TPL) and 5-fluorouracil(5-FU) on hepatocellular carcinoma (AsPC-1) cells growth and apoptosis and explore the synergy of 5-fluorouracil combined with triptolide in AsPC-1 cells . To investigat whether triptolide could enhance 5-fluorouracil-induced apoptosis in AsPC-1 cells or not. Method AsPC-1 cells were cultured in the culture medium with high glucose DMEM and 10% fetal calf serum. Inhibited proliferation effects of 5-fluorouracil, triptolide, IC30 dose of triptolide combined with 5-fluorouracil and 5-fluorouracil combined with triptolide on AsPC-1 cells was detected by MTT assay. The expression changes of PARP and caspase-3 were detected by Western Blot.The cell apoptosis ratio in each experimental group was analyzed by flow cytometer. Result The proliferation of AsPC-1 cells could be inhibited by triptolide and 5-fluorouracil with dose and time related. The 48h IC50 of 5-fluorouracil on AsPC-1 cells was 9.42±1.87ug/ml and its 72h IC50 was 2.24±0.44ug/ml. The 48h IC50 of triptolide on AsPC-1 cells was 13.75±1.25ng/ml and its 72h IC50 was 2.60±0.33ng/ml . The 72h IC30 of triptolide on AsPC-1 cells was 1.47±0.29ng/ml. IC30 concentration of triptolide could significantly improve the proliferation inhibition of 5-fluorouracil on AsPC-1 cells.When we compared the the IC50 concentration of combined drug group with 5-fluorouracil single drug group, we found that the difference was statistical significance (P<0.05). Triptolide combined with 5-fluorouracil on AsPC-1 cells had a strong synergistic effect. After 24 hours, we found that the expression amount of activated caspase3 in 5-fluorouracil alone group was much more than other groups,with significant difference (P <0.01). After 48 hours, the expression amount of activated caspase3 in combined drug group was much more than single drug group and control group, with statistically significant difference (P <0.01). After 48 hours, the expression amount of activated PARP in combined drug group was much more than single drug group and control group,with statistically significant difference (P <0.01). Compared combined drug group with single drug group and control group.We found that the proportion of cells in combined drug group was higher than other three groups , with statistically significant differences (P <0.01). Conclusion Triptolide had significant effects of inhibiting proliferation and inducing apoptosis on AsPC-1 cells and no significantly inhibitory effect of IC30 concentration of triptolide could significantly enhance proliferation inhibition of 5-fluorouraci on AsPC-1 cells. 5-fluorouraci could inhibit the proliferation and induce apoptosis to a certain extent on AsPC-1 cells. Triptolide combined with 5-fluorouraci had highly synergistic effect on inhibiting proliferation of AsPC-1 cells . Triptolide could obviously enhance 5-fluorouracil-induced apoptosis in AsPC-1 cells.