Effect Research On Three-dimensional Multicellular Spheroids Models Of Colon And HT-29 Xenograft In Nude Mice Of Petroleum Ether Part Form Anthriscus Sylvestris Combined With Fluorouracil

Objective:To investigate the effects of petroleum ether part form Anthriscus sylvestris combined with fluorouracil and the mechanism on DLD-land HCT116 three-dimensional multicellular spheroids models through the cell test and investigate the inhibitory effect of HT-29 xenograft in Nude Mice,for expounding about the anti-tumor mechanisms of petroleum ether part form Anthriscus sylvestris as a mixture and the basic research data of the development of New Drugs against colon carcinoma.Methods:(1)to test the inhibition effect of the petroleum ether part from Anthriscus sylvestris respectively for the Flat cells and the multi-cellular spheroids model of DLD-1 and HCT116 colon cancer cell by CCK8 method and to investigate the inhibition effect of dosage of the petroleum ether part from Anthriscus sylvestris combined with fluorouracil;(2)to test the petroleum ether part from Anthriscus sylvestris combined with the fluorouracil in the influence of the cell cycles and apoptosis of the DLD-1 and HCT116 multi-cellular spheroids model by the flow cytometry and to investigate the effect of dosage of the petroleum ether part from Anthriscus sylvestris combined with fluorouracil;(3)to detect the petroleum ether part from Anthriscus sylvestris combined with the fluorouracil in the expression of the DLD-1 and INOS of the DLD-1 and HCT116 multi-cellular spheroids model by western blot;(4)to test the inhibition effect of the petroleum ether part from Anthriscus sylvestris combined with the fluorouracil respectively for the Flat cells and the multi-cellular spheroids model of HT-29 colon cancer cell by CCK8 method through the cell text and inhibitory effect of HT-29 xenograft in Nude Mice by observing the mice weight and tumor volume.Results:(1)the petroleum part from Anthriscus sylvestris could inhibit the DLD-land HCT116 two-dimensional cellular and three-dimensional multi-cellular spheroids,two-dimensional cellular of DLD-1 IC50=56.67μg/ml,three-dimensional multi-cellular spheroids of DLD-1 IC50=89.78μg/ml,two-dimensional cellular of HCT116 IC50=35.79μg/ml,three-dimensional multi-cellular spheroids of HCT116 IC50= 60.59μg/ml,and the effect in dose-dependent manner to some extent;(2)the petroleum part from Anthriscus sylvestris combined with the 5-FU could significantly improve the inhibition effect of the DLD-1 and HCT116 three-dimensional multi-cellular spheroids;(3)5-FU was blocked in the S phases of the DLD-1 and HCT116,the petroleum part from Anthriscus sylvestris was blocked in the G0/G1 phases of DLD-1 and in the G2/M phases of HCT116;to conduct the combined apply of two kinds of the drugs,the proportion in the S phase and GO/Glor G2/M phases would increased;(4)both the petroleum part from Anthriscus sylvestris and the 5-FU could improve the apoptosis of the DLD-1.and HCT 116 conduct the combined apply of two kinds of the drugs,the apoptosis rate would significantly increased;(5)the petroleum part from Anthriscus sylvestris combined with the 5-FU could significantly inhibit the expression of the p-AKT protein and INOS protein in the DLD-1 and HCT116;(6)the petroleum part from Anthriscus sylvestris combined with 5-FU can inhibit the growth of HT-29 xenograft.Conclusion:(1)the petroleum part from Anthriscus sylvestris are disincentive for DLD-1 and HCT116 Flat two-dimensional cellular and three-dimensional multi-cellular spheroids,and the effect in dose-dependent manner to some extent;(2)The inhibitory effect of the petroleum part from Anthriscus sylvestris combined with 5-FU on DLD-1 and HCT116 cells resulted the change of cell cycle blocking and cell apoptosis.This effect may be related to p-AKT and INOS proteins;(3)the petroleum part from Anthriscus sylvestris combined with 5-FU can inhibit the growth of HT-29 xenograft.