Effects and molecular mechanism of miR-21 on cell invasion, migration and apoptosis of pancreatic cancer PANC1 cells / 中华胰腺病杂志

ABSTRACT

Objective:To observe the effects of microRNA-21 (miR-21) on cell invasion, migration and apoptosis of pancreatic cancer PANC1 cells, and explore the potential molecular mechanism.Methods:Recombinant plasmids carrying miR-21 or small interfering RNA targeting miR-21 were constructed. Using blank plasmid as the control, the recombinant plasmids were transfected with human pancreatic cancer PANC1 cells by liposome method, respectively to establish blank group, miR-21 overexpression group (overexpression group) and miR-21 silence group (silence group) PANC1 cell lines. Cell proliferation, apoptosis, invasion and metastasis were detected by MTT method, flow cytometry, transwell chamber and wound scratch test, respectively. ELISA and Western blot were used to measure the protein expression of programmed cell death factor 4(PDCD4), gene of phosphate and tension homology deleted on chromsome ten (PTEN), vascular endothelial growth factor (VEGF), survivin, matrix metalloproteinase 2 (MMP-2) and MMP-9.Results:After 24 h cell culture, the cell proliferation rate of blank group, overexpression group and silence group was (20.72±5.62)%, (28.46±6.12)% and (14.05±3.36 )%; cell apoptosis rate was (5.89±0.26)%, (4.62±0.19)% and (8.66±2.55)%; the number of transmembrane cells was (212.4±32.5), (508.8±50.7) and (50.9±10.6) per 200 times visual field; the area covered by migrated cells was (75.6±12.1), (118.8±20.2) and (48.8±9.5)mm 2 per 200 times visual field; the expression of PDCD4 was 0.85±0.22, 0.72±0.10 and 1.36±0.41; the expression of PTEN was 0.85±0.21, 0.28±0.09 and 1.36±0.45; the expression of VEGF was 0.79±0.24, 1.15±0.31 and 0.46±0.10; the expression of survivin was 1.02±0.33, 1.51±0.42 and 0.52±0.12; the expression of MMP-2 was 1.12±0.22, 1.86±0.52 and 0.56±0.18; the expression of MMP-9 was 1.06±0.15, 1.78±0.48 and 0.49±0.12. All the differences among the three groups were statistically significant (all P<0.05). Compared with blank group, the cell apoptosis rate, PDCD4 and PTEN expression were increased, while cell proliferation, invasion and migration, and the protein expression of VEGF, survivin, MMP-2 and MMP-9 were all decreased; the changes in silence group was totally contrary to those in overexpression group. All the differences among the three groups were statistically significant (all P<0.05). Conclusions:miR-21 silencing can inhibit the proliferation, migration and invasion of PANC1 cells and promote apoptosis, and the mechanism was possibly associated with the upregulation of PDCD4 and PTEN protein expression.