Objective To investigate whether curcumin could inhibit the metastasis of pancreatic cancer in vitro and in vivo and its mechanism.Methods Pancreatic cancer AsPC-1 cells were treated with different concentrations of curcumin.After 72 hours,cell survival rate were detected by MTT and the appropriate concentration of curcumin was selected.After treated with 6 μmol/L curcumin,early apoptosis of AsPC-1 cells were detected by Annexin V-FITC/PI double staining flow cytometry,and the effects of curcumin on the migration and invasion of AsPC-1 cells were observed.After establishment of the orthotopic nude mouse model of pancreatic cancer,the mice were orally treated with low dose (20 mg/kg body weight) and high dose (40 mg/kg body weight) of curcumin respectively.Eight weeks later mice were sacrificed to observe the tumor metastasis,immunohistochemical methods were used to detect the positive expressions of CD34,NF-κB and MMP-9 in the tumor tissue.Results Curcumin of different concentrations could inhibit the proliferation of AsPC-1 cells and induce its apoptosis in a dose dependent manner,and 6.μmol/L was the best dose of curcumin.After 6 μmol/L curcumin treatment,the migration coverage of AsPC-1 decreased from 70％ to 10％,the number of penetrating cells decreased from 136/200x magnification to 17/200x magnification,and the difference between the two groups was statistically significant (P ＜0.05).In the nude mouse model of pancreatic cancer,the size of the tumor decreased from (97.8 ± 15.3) mm3 to (44.3 ± 9.7) mm3 in high dose group and (28.1 ±7.1)mm3 in low dose group,the number of distant metastasis decreased from 108.3 ±6.7 to 29.5 ±4.5 and 8.9 ± 2.4,the expression of CD34 decreased from 20.5 ± 2.3 to 10.3 ± 1.2 and 7.9 ± 3.2,and the expression of MMP-9 decreased from 85.2 ± 2.3 to 53.2 ± 1.2 and 34.2 ± 3.1,and the difference was statistically significant (P ＜ 0.05).Conclusions Curcumin can inhibit the metastasis of pancreatic cancer both in vitro and in vivo,and its effect may be related to the inhibition of expressions of CD34 and MMP-9.

Objective To investigate the effect and mechanism of oxymatrine on invasion and metastasis of human pancreatic cancer line SW1990 in vitro. Methods Human pancreatic cancer cell line SW1990 was cultured in vitro. Oxymatrine was added into the culture media of SW1990 cells. Then MTT assay was used to determine the effect on proliferation. The adhesive capability, the mobile ability and invasive ability of SW1990 cells were detected by the adhesion assay, the crossing-river test, the transwell migration assay and the matrigel invasion method, respectively. RT-PCR was used to detect the mRNA expression of MMP-2 and VEGF. ELISA method was used to detect the protein levels of the VEGF. Results The growth of SW1990 cells was inhibited by oxymatrine in a dose and time-dependent manner. After 2 mg/ml of oxymatrine treatment for SW1990 cells for 1 h, the adhesive capability inhibitory rate was (35.23 ±8.56) % ; 24 h later, crossing-river time was (65.46 ±4.25) h, which was significantly longer than that in control group [ (34.50 ± 4.12) h, P <0.05)], the number of penetrating cells was 91.9 ±9.6, which was significantly lower than that in control group (144.2±17.2, P <0.05). The mRNA expression of MMP-2 and VEGF, expression of protein of VEGF in SW1990 cells was significantly down-regulated [0.515 ±0.063 vs. 0.817 ±0.054, 0.343 ± 0.072 vs. 0.650 ±0.068; (265.50 ±5.45) pg/ml vs. (441.06 ±16.70) pg/ml, P <0.05]. Conclusions Oxymoron can inhibit the invasion and metastasis ability of pancreatic cancer line SW1990 in vitro, and the mechanism is possibly related to the down-regulation of MMP-2 and VEGF expression.

Objective To explore the influence of LPS treatment on related molecules in Smads and ERK1/2 signal pathway in pancreatic stellate cell line LTC-14.Methods LTC-14 cells were cultured in vitro, and were treated with LPS at different dose in different time points.Protein expressions of related molecules in Smads pathway and ERK1/2 pathway and α-SMA in LTC-14 Cells were examined by Western blot.Results On Treated LTC-14 cells by 0, 1, 5, 10, 20 and 50 mg/L LPS,protein expressions of Smad3 were 0.15±0.02, 0.37±0.02, 0.44±0.01, 0.46±0.02, 0.372±0.01 and 0.24±0.03;expressions of Smad7 were 0.79±0.05, 0.84±0.02, 0.55±0.03, 0.45±0.03, 0.34±0.02 and 0.92±0.07;p-ERK1/2 levels were 0.48±0.05, 0.74±0.03, 0.72±0.04, 0.89±0.02, 0.81±0.02 and 0.72±0.03;p-cPLA2 levels were 0.15±0.03, 0.30±0.01, 0.31±0.01, 0.30±0.02, 0.28±0.03 and 0.32±0.02;α-SMA levels were 0.56±0.06, 0.62±0.06, 0.54±0.04, 1.03±0.11, 1.39±0.08 and 1.28±0.10.The changes of protein expressions before and after LPS treatment were obvious (all P<0.01).The protein expressions of ERK1/2 were 0.56±0.03, 0.57±0.02, 0.53±0.02, 0.58±0.02, 0.59±0.05 and 0.55±0.04, which did not change obviously along with increased LPS dosages.LTC-14 cells treated with 10 mg/L LPS for 0, 1, 3, 6 and 9 h,the expressions of Smad3 were 0.69±0.05, 0.68±0.07, 1.02±0.14, 1.82±0.0 and 2.04±0.11,those of Smad7 were 2.77±0.10, 1.37±0.08, 1.45±0.14, 0.78±0.09 and 0.63±0.06,those of p-ERK1/2 were 0.16±0.03, 0.32±0.05, 0.79±0.03, 1.50±0.07 and 1.77±0.04,those of p-cPLA2 were 0.15±0.04, 0.32±0.06, 0.63±0.04, 0.95±0.04 and 1.49±0.10,those of α-SMA were 0.84±0.03, 1.26±0.21, 1.81±0.19, 4.28±0.26 and 4.37±0.15, all of which changed obviously as the treatment time increased (P<0.05 or 0.01).The expressions of ERK1/2 were 0.75±0.03, 0.72±0.02, 0.80±0.04, 0.74±0.03 and 0.85±0.09, which did not change obviously as the treatment time increased.Conclusions LPS could upregulate the expression of α-SMA in a time-and dose-dependent way, and activate intracellular Smads and ERK1/2 inflammatory pathways, which may be the potential molecular mechanism of the development of chronic pancreatitis.