Expression And Significance Of Axin2 In Pancreatic Carcinoma

Objective:To investigate the expressions of Axin2 in pancreatic cancer cells and to observe the influence of Axin2 on the ability of proliferation, invasion and migration of human pancreatic cancer cells(PANC-1).To expound the role of Axin2 in the occurrence and development of pancreatic cancer and to provide theoretical basis for pancreatic gene targeted therapy and development of anti-tumor drugs.Methods:1. The expressions of Axin2 in pancreatic cancer cell line with different invasive ability(Bx PC-3<Mia Pa Ca-2<PANC-1) and immortalized normal pancreatic cells(H6C7) were detected by QPCR, Pancreatic cancer cell line with the lowest expressions of Axin2 was screened, 2. Plasmid that over expressing Axin2 was constructed and divided into three groups: the negative control group, the blank control group and the over-expressions group. The over-expressions group and the negative control group were successfully transfected with over-expressed-Axin2 plasmid vectors and blank vectors, while the blank control group was not transfected, QPCR was used to detect the expressions of Axin2 in each group after transfection, 3.Pancreatic cancer cells with stable expressions of Axin2 after transfection were screened and grouped rationally, and then an in vitro functional experiment was carried out on them. MTT assay, Transwell assay and scratch test were used to determine the cell's proliferation, invasion and migration ability after transfection with over-expressed Axin2.Results:1. QPCR results showed that: the expressions of Axin2 in normal pancreatic cell lines and three different invasion and metastasis of pancreatic cancer cell lines were determined. The relative expression level of Axin2 in PANC-1, Bx PC-3, Mia Pa Ca-2 and normal pancreatic cell H6C7was(0.13±0.01),(0.42±0.05),(0.24±0.01) and(1.01±0.01), P<0.05 for the difference was statistically significant, 2. Pancreatic cancer cells with the lowest expressions were screened. Transfection efficiency of PANC-1 cell after transfecting the vector that over expressing Axin2 was detected by QPCR. The results indicate that relative expression level of Axin2 in over expression group, negative control group and blank group was(4.32±0.07),(1.09±0.08) and(0.92±0.08). There were no significant differences between blank group and negative control group(P>0.05). However, compared to blank group and negative control group, expression level in over expression group was significant higher(P<0.05), 3. Pancreatic cancer cells with stable expressions of Axin2 after transfection were screened and then an in vitro functional experiment was carried out on them. 1). MTT showed the relative growth rates were(0.33±0.02),(0.99±0.05) and(1.00±0.22) in the over-expression group, the negative control group and the blank control group, respectively, the growth rate of the over-expression group was significantly lower than those of the other two groups, with statistically significant differences(P<0.05), 2). Scratch test showed compared with the blank control group and the negative control group, migration ability of PANC-1 cells was significantly reduced in the over-expression group, 3).Transwell assay showed compared with the blank control group and the negative control group, invasion ability of PANC-1 cells was significantly declined in the over-expression group.Conclusions:1. Expressions of Axin2 is down-regulated in pancreatic cancer cell lines than in the normal pancreas cells, which is lower in cancer cells with higher invasive ability. 2. High expression of Axin2 can decrease the proliferation, invasion and migration ability of PANC-1 cells, promoting a role of tumor-suppressor gene.