Effects of RegIV on proliferation, invasion, and migration of pancreatic cancer cells PANC-1

HU Qil; LI Jian; MENG Hongbo; ZHOU Bo; WANG Feng; SONG Zhenshun

doi:10.3969/j.issn.1002-266X.2017.29.001

Abstract：Objective To explore the effects of RegIV gene on the proliferation, invasion, and migration of pancreatic cancer cells PANC-1.Methods PANC-1 cells in logarithmic growth phase were cultured in the incubator until they reached 70%-80% confluence.PANC-1 cells were transfected with Puro-P-RegIV-shRNA lentiviral vector (P-RegⅣ-shRNA group) and Puro-shRNA lentiviral empty vector (P-NC-shRNA group), respectively.Cells without transfection or with null-transfection were served as controls (PANC-1 group).At 48 h after transfection, quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the expression of RegIV mRNA and protein.The CCK-8 test was used to detect the proliferation.Scratch test and Transwell chamber test were used to detect the migration and invasion of the cells, respectively.Ten 5-week-old female nude mice were randomly divided into the P-RegⅣ-shRNA nude mouse group and P-NC-shRNA nude mouse group with 5 in each group, which were then injected with P-RegⅣ-shRNA and P-NC-shRNA cells in logarithmic growth phase in the above two groups.The growth of tumor nodules was observed every day from the day of inoculation.After 6 weeks, the nude mice were sacrificed and the tumor volume was measured.Results The expression levels of RegIV mRNA and protein in the P-RegⅣ-shRNA group were significantly lower than those of the PANC-1 group and P-NC-shRNA group (both P<0.05).Cells in the P-RegⅣ-shRNA group had a lower proliferative capacity than cells in the P-NC-shRNA group and PANC-1 group (P<0.05).Moreover, compared with the PANC-1 group and P-NC-shRNA group, Puro-P-RegIV-shRNA group showed shorter transmembrane distance and decreased number of transmembrane cells, indicating decreased migration ability (all P<0.05).No significant differences were found in the above parameters between the PANC-1 group and P-NC-shRNA group (all P>0.05).Besides, tumor growth rate and the tumor volume in nude mice were decreased significantly in the P-RegⅣ-shRNA nude mouse group as compared with those of the P-NC-shRNA nude mouse group (P<0.01).Conclusion Down-regulation of RegIV could inhibit the proliferation, invasion, and migration of PANC-1 cells.