Effects of Aqueous Extract of Clove on Proliferation, Migration and Invasion of Pancreatic Cancer Cells

OBJECTIVE To investigate the effects of aqueous extract of clove(AEC) on proliferation and apoptosis of the human pancreatic cancer cells Panc-1 and Panc-28, and preliminarily clear the molecular mechanisms. METHODS MTT assay was used to detect the proliferation of Panc-1 and Panc-28 cells treated with AEC respectively, and flow cytometry was used to detect cells apoptosis, and clonal test was used to detect cells colony formation rate, and transwell assay was performed to detect cells migration and invasion. The level expression of cleaved-PARP, Bax, Bcl-2, E-cadherin protein were determined by Western blotting. RESULTS The results of MTT experiments revealed that 25 μg·mL^-1 of AEC was able to inhibit Panc-1 and Panc-28 cells growth compared with the control group at 72 h, and the inhibition rate reached (16.21±3.57)% and (22.44±4.92)% (P<0.01), respectively. The flow cytometry results indicated 200 μg·mL^-1 of AEC significantly induced Panc-1 and Panc-28 cells apoptosis compared with the control group at 24 h; the early apoptotic rates were (11.52±0.34)% and (30.88±0.73)%(P<0.01); the late apoptotic rates were (23.59±1.04)% and (13.27±0.85)%(P<0.01) respectively. The experiment of clonal formation ability showed 50 μg·mL^-1AEC could inhibit the formation of Panc-1 and Panc-28 cells colonies(P<0.01). Transwell assay revealed Panc-1 and Panc-28 cells were treated with 100 μg·mL^-1AEC for 48 h, respectively, the migration cells number decreased from 275±7 to 165±15, and 227±25 to 124±16(P<0.01); the invasion cell number decreased from 179±19 to 106±7 and 132±7 to 61±5 (P<0.01). Western blotting results confirmed that 100 μg·mL^-1 AEC up-regulated the expression of cleaved-PARP(P<0.01) and E-cadherin(P<0.05), and increased the ratio of Bax/Bcl-2(P<0.01) in Panc-1 and Panc-28 cells. CONCLUSION AEC can induce the apoptosis of Panc-1 and Panc-28 cells, and inhibit their proliferation, clone formation, migration and invasion, speculating that its mechanism may be related to the upregulation of E-Cadherin expression and increasing of ratio of Bax/Bcl-2.