|
Objective:To explore the effects of aspirin and curcumin on COX-2,/3-catenin and PGE2 expressions in human colon cancer cell lines of HT-29 and SW480 cells and to provide the theoretical and experimental basis for clinical treatment of colon cancer.Methods:MTT was employed to detect the influence of aspirin and curcumin on cell growth proliferation on the in vitro cultured human colon HT-29 and SW480 cell lines. ELISA was used to detect PGE2 protein levels changes after drug treatment on tumor cells, Real-time PCR was applied to assay the COX-2 andβ-catenin gene mRNA expression levels between drugs-treated and untreated cells, and the protein expression levels were analyzed by Western blot.Results:The IC50 of aspirin for HT29 and SW480 was 7.9 mM and 8.12 mM, The IC50 of curcumin for HT 29 and SW480 was 19.27μM and 49.5μM, respectively. The Inhibition rates of SW480 were 11.89±0.241,37.99±0.076,64.21±0.034,70.46±0.029 and 57.82±0.099 after treated by curcumin at12 h,24 h,36 h,48h and 60h. The Inhibition rates of HT-29 were 8.04±0.216,22.58±0.063,57.10±0.050,77.83±0.034 and 74.88±0.015 after treated by curcumin 12 h,24 h,36 h,48h and 60h. In the same case, the Inhibition rates of HT-29 were 24.07±0.204,25.82±0.210,54.36±0.027, 63.00±0.023 and 69.99±0.009 after treated by aspirin 12 h,24 h,36 h,48h and 60h. Both aspirin and curcumin were all showed a time-effect relationship, in such two cells. In HT-29 cells, the mRNA expression level of COX-2 was down-regulated to 0.6,0.56 and 0.91 times in cells after treated at 6h,12h and 24h by curcumin, and the mRNA expression level ofβ-catenin was down-regulated to 0.4,0.95 and 0.48 times, (P<0.05, respectively). The mRNA expressions of COX-2 were down-regulated to 0.79,0.52 and 0.57 times in cells after treated by aspirin at 6h,12h and 24h and the mRNA expressions ofβ-catenin were down-regulated to 0.81,0.66 and 0.95 times, (P<0.05, respectively). Meanwhile, in SW480 cells, contrast to the blank samples, the mRNA expressions of COX-2 were down-regulated to 0.62,0.3 land 0.11 time in cells after treated 6h,12h and 24h by curcumin, and the mRNA expressions ofβ-catenin were down-regulated to 0.4,0.95 and 0.48 time(P<0.05, respectively). The mRNA expressions of COX-2 were down-regulated to 0.63,0.35 and 0.1 time in cells after treated 6h,12h and 24h by aspirin and the mRNA expression levels ofβ-catenin were down-regulated to 0.81,0.79and 0.79 time(P<0.05, respectively). In the HT-29cells, PGE2 levels were 15.89095μg/L,26.0316μg/L and 59.20192μg/L,β-catenin /β-actin were 0.285±0.098,0.225±0.092 and 0.362±0.089, and the COX-2/β-actin were 0.352±0.053,0.302±0.083and 0.194±0.092 after treated 6h,12h and 24h by asprin. PGE2 levels were 18.44981μg/L,40.05791μg/L and 86.0225μg/L,β-catenin /β-actin were 0.256±0.079,0.219±0.095 and 0.215±0.083, and COX-2/β-actin were 0.219±0.087,0.382±0.069 and 0.185±0.079 after treated 6h,12h and 24h by curcumin contrast to blank samples which were 29.19226,57.90828 and110.7913 treated 6h,12h and 24h without any drug(P<0.05, respectively).In the SW480 cells, PGE2 levels were 55.12671μg/L,78.06162μg/L and 143.6441μg/L,β-catenin /β-actin was 0.206±0.078,0.144±0.079 and 0.062±0.079, and the COX-2/β-actin was 0.352±0.053,0.302±0.083 and 0.194±0.092 after treated 6h,12h and 24hby asprin. PGE2 levels were 23.2832μg/L,31.3388μg/L and 525.17865μg/L,β-catenin /β-actin was 0.251±0.097,0.216±0.095 and 0.141±0.059, after treated 6h,12h and 24h by curcumin contrast to blank samples which protein level was 61.93492,85.39108 and 170.1176 after treated 6h,12h and 24h without any drug(P<0.05, respectively).Conclusions:The hibitory rates of proliferation treated by aspirin or curcumin on cultured human colon cancer HT-29 and SW480 cell were in a time-and-dose dependent manner. Aspirin inhibited the growth of HT-29 cell through inhibited the mRNA and protein expressions of COX-2 andβ-catenin, reduced the PGE2 contents. Curcumin inhibited the growth of HT-29 cell also through inhibited the mRNA and protein expressions of COX-2 andβ-catenin, reduced the PGE2 products.
|