Influence of microRNA-135a on proliferation and apoptosis of human pancreatic cancer cells

Abstract

Objective To investigate the influence of miRNA-135a on the proliferation and apoptosis of the pancreatic cancer cells.Methods Methyl thiazol tetrazolium (MTT) was used to detect the inhibitory effect of 5-fluorouracil (5-Fu) on Bxpc-3 cells and the the appropriate concentration of 5-Fu was chosen in the following experiment.The apoptosis of Bxpc-3 cells was detected by using flow cytometry.Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the mRNA expression level of miRNA-135a after the Bxpc-3 cells were treated with 5-Fu.The mimic and anti-miRNA oligonuclotide (AMO) were designed and transfected into Bxpc-3 cells,and the Bxpc-3 cells transfected with vehicle served as control group.FQ-PCR was used to detect the expression of miRNA-135a and the proliferation rate was detected by using MTT assay.Flow cytometry was used to detect the apoptosis of transfected Bxpc-3 cells after treatment with 5-Fu.Results The 5-Fu inhibited Bxpc-3 cells in a concentration-and time-dependent manner.With the increase of concentration and prolongation of time,the inhibition rate of 5-Fu was elevated.The miRNA-135a level was reduced to (0.45 ±0.02),(0.60 ±0.03)and (0.73 ± 0.02) times after treatment with 5-Fu (P ＜ 0.05 for all).After transfection the mRNA levels of miRNA-13a was elevated (9.73 ±0.83) times in the mimic groups and reduced to (0.26 ±0.01) times in the AMO groups as compared with the vehicle groups (P ＜ 0.05 for all).The proliferation of Bxpc-3 cells was elevated in the mimic group and reduced in the AMO group as compared with the vehicle group.The apoptosis of Bxpc-3 cells,which was (12.03 1.07) ％,was reduced significantantly in the mimic groups as compared with the vehicle groups [(20.61 ± 1.53) ％] (P ＜ 0.05),while the apoptosis was elevated in the AMO groups [(38.20 ±2.64)％] (P ＜0.05).Conclusion The level of miRNA-135a was lower in Bxpe-3 cells treated with 5-Fu.The up-regulated expression of miRNA-135a may promote the proliferation and reduce the apoptosis rate of Bxpc-3 cells.The down-regulated miRNA-135a may inhibit the proliferation and augment the apoptosis of Bxpc-3 cells. Key words: Pancreatic cancer; MicroRNA ; Apoptosis