Effect of miRNA-221-3p on apoptosis of pancreatic cancer cells and its mechanism / 肿瘤研究与临床

ABSTRACT

Objective:To investigate the expression of miRNA-221-3p (miR-221-3p) in pancreatic cancer cells and its effect on apoptosis of pancreatic cancer cells, and the possible related mechanisms.Methods:Pancreatic cancer cell line PATU8988T was selected and transfected with miR-221-3p mimics, miR-221-3p inhibitors and their corresponding negative control sequences using Lipofectamine 3000. PATU8988T cells were divided into negative control group (without any treatment), miR-221-3p mimics negative control group, miR-221-3p mimics group, miR-221-3p inhibitors negative control group, and miR-221-3p inhibitors group. Real-time quantitative fluorescence polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of miR-221-3p, flow cytometry was used to detect the influence of miR-221-3p on apoptosis of pancreatic cancer cells, and Western blotting was used to detect the expressions of P53 and PTEN proteins in PATU8988T cell line.Results:The relative expression levels of miR-221-3p in negative control group, miR-221-3p mimics negative control group, miR-221-3p mimics group, miR-221-3p inhibitors negative control group and miR-221-3p inhibitors group were 1.02±0.18, 1.50±0.33, 2.96±0.70, 1.62±0.30, and 0.36±0.05, respectively, and the difference was statistically significant ( F = 12.61, P < 0.05); the relative expression level of miR-221-3p in miR-221-3p mimics group was higher than that in negative control group and miR-221-3p mimics negative control group ( t = 1.94, P < 0.05; t = 1.45, P < 0.05); the relative expression level of miR-221-3p in miR-221-3p inhibitors group was lower than that in negative control group and miR-221-3p inhibitors negative control group ( t = -0.65, P < 0.05; t = -1.26, P < 0.05). The apoptosis rates in negative control group, miR-221-3p mimics negative control group, miR-221-3p mimics group, miR-221-3p inhibitors negative control group and miR-221-3p inhibitors group were (8.60±0.20)%, (8.60±0.26)%, (4.27±0.31)%, (8.83±0.29)%, and (13.63±0.60)%, respectively, and the difference was statistically significant ( F = 253.80, P < 0.01); the apoptosis rates in miR-221-3p mimics group was lower than that in negative control group and miR-221-3p mimics negative control group ( t = -4.33, P < 0.05; t = 4.33, P < 0.05); the apoptosis rate in miR-221-3p inhibitors group was higher than that in negative control group and miR-221-3p inhibitors negative control group ( t = 5.03, P < 0.05; t = 4.80, P < 0.05). There was no statistical difference in expression levels of P53 and PTEN proteins between miR-221-3p mimics negative control group and miR-221-3p inhibitors negative control group (P53 t = 0.22, P > 0.05; PTEN t = 0.33, P > 0.05); the expression levels of P53 and PTEN proteins in miR-221-3p mimics group were decreased compared with the miR-221-3p mimics negative control group (P53 t = 4.31, P < 0.05; PTEN t = 8.49, P < 0.05); the expression levels of P53 and PTEN proteins in miR-221-3p inhibitors group were increased compared with the miR-221-3p inhibitors negative control group (P53 t = 5.17, P < 0.05; PTEN t = 6.21, P < 0.05). Conclusions:miR-221-3p is highly expressed in pancreatic cancer PATU8988T cells, which can inhibit the apoptosis of pancreatic cancer cells. miR-221-3p may regulate the progression of pancreatic cancer through P53 and PTEN.