The Effects Of 5-ally-7-gen-difluormethylenechrysin On Growth And Expression Of Tumor-related Gene In SMMC-7721

Objective: In order to find some new candidates for tumor chemotherapy, we investigate the growth proliferation inhibitition of SMMC-7721 induecd by 5-ally-7-gen-difluormethylenechrysin (ADFMChR), and whether the mechanism related to the activation of PPARγ, and analysis tumor-related genes expression change induced by ADFMChR in SMMC-7721 cells.Methods: SMMC-7721 cells was cultured in vitro. The trypan blue exclusion methods was used to evaluated the effect of ADFMChR,GW99662 and combination of both on proliferation in SMMC-7721 cells. The anchoring capacity of SMMC-7721 cells treated by ADFMChR, GW99662 and combination of both was observed by plate clone. Soft agar was used to measure the foci forming ability of SMMC-7721 cells treated by ADFMChR, GW99662 and combination of both. The expression profile of cancer-related genes induced by ADFMChR were explored by gene array.Results: Trypan blue exclusion method shown that ADFMChR of various concentration 1.0, 3.0, 10.0μM all possess the growth inhibitory effect in SMMC-7721 cells was treated 48h, when the concentration was added from 1.0μM to 10.0μM, the doubling time of SMMC-7721 cells was extended from 29.41h to 48.89h, and preincubating SMMC-7721 cells with 10.0μM GW9662 could significantely cut down the growth inhibitory effect of ADFMChR. The plate clone shown that the foci inhibitory rate of ADFMChR of various concentration 1.0, 3.0, 10.0μM in SMMC-7721 cells were 10.41%±3.67 % , 28.01%±2.40 % , 47.21%±8.66 % respectively, but the foci inhibitory rate of 3.0μM ADFMChR in SMMC-7721 cells preincubated by 10μM GW9662 was 15.21±6.04%, which is obviously lower than the value of 3.0μM ADFMChR(P < 0. 05). The sofr agar foci formation assay shown that the concentration increase of ADFMChR was followed by the decrease of average foci number. The average foci number of 3.0μM ADFMChR in SMMC-7721 cells preincubated by GW9662(10.0μM) significantly higher than the value of 3.0μM ADFMChR (P < 0. 05). GeneArray indicated that 6 genes up-regulated in SMMC-7721 cells treated 4h with 3.0μM ADFMChR, including LCN2, MAPK13, MMP14, MMP17, MMP2, TFDP2, and 23 genes down-regulated, including ABCB1, ANPEP, RHOD, BHLHB2, CDKN1C, CRHR1, CTSD, DHRS2, FOXO1A, GPR39, ID1, JAK1, PCTK1, PCTK2, ZMYND8, SEMA3C, SMPD1, SPINT2, SRC, STAT2, TRRAP, UCHL1, VHL.Conclusion:1) ADFMChR could inhibit the growth of SMMC-7721 cells in a dose-dependent manner, the mechanism could related with activation of PPARγ.2) The expression profile of cancer-related genes in SMMC-7721 cells suggested that ADFMChR up-regulate MAPK13 and down-regulate FOXO1A,GPR39,TRRAP,SEMA3C,SRC,ANPEP,CTSD and ID1, which could related with the growth inhibition of SMMC-7721 cells.