Influence of microRNA-135a on proliferation and apoptosis of human pancreatic cancer cells
Tao Qin, Qiang Fu, Chuanjiang Liu, Mingxing Hu, Qiang Tang, Yuzhu Wang, Fei Xue
Abstract
Tao Qin, Qiang Fu, Chuanjiang Liu, Mingxing Hu, Qiang Tang, Yuzhu Wang, Fei Xue
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
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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
Key concepts: Flow cytometry, Apoptosis, Transfection, MTT assay, Molecular biology, Pancreatic cancer, microRNA, Messenger RNA