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Expression of miR-223 in pancreatic cancer cells and influence on proliferation, apoptosis and cell cycle

Author: ZuoZhongXing
Tutor: ZhaoYuPei
School: Beijing Union Medical College
Course: Clinical
Keywords: miR-223 pancreatic cancer target gene cell proliferation apoptosis cell cycle
CLC: R735.9
Type: Master's thesis
Year: 2011
Downloads: 13
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Abstract


Objective:To analyze miR-223expression in pancreatic cancer cell lines and investigate the impact of miR-223on cell proliferation, cell apoptosis and cell cycle of the pancreatic cancer cells as well as the mechanisms of action.Methods:We analyze miR-223expression in pancreatic cancer cell lines by real-time PCR. Subsequently we employed a bio-computational approach to identify miR-223target gene followed with dual luciferase reporter assay validation. Finally we transfected T3M4pancreatic cancer cells with miR-223mimics and transfected SW1990pancreatic cancer cells with miR-223inhibitor to investigate the impact of miR-223on the biological functions. Cell proliferation was detected with CCK-8; Cell apoptosis was analyzed with Annexin V/PI assay; Cell cycle distribution was analyzed with flow cytometry.Results:miR-223expression was detected in all8pancreatic cell lines possessed by our laboratory, the expression in Panc-1, BxPC-3, Capan-1, COLO357, SW1990and MiaPaCa-2is relatively high, well the expression in AsPC-1and T3M4is relatively low; Then we utilized the algorithms Target Scan, Miranda, Diana-MicroT, PicTar etc. to predict the target gene of miR-223, candidate target gene FOXOla was chosen. Dual luciferase reporter assay was then carried out and show that compared with the positive control group, there was a decrease in the fluorescence intensity and the decrease had statistically difference (P＜O.05); Down-regulation of miR-223in SW1990lead to decreased proliferation, reduction in the proportion of cells in the S phase., but found no regular change in Gl and G2/M phase; on the other hand, up regulation of miR-223in T3M4lead to increased proliferation, increase in the proportion of cells in the S phase with a concomitant reduction in Gl phase, but found no regular change in G2/M phase; up and down regulation of miR-223in this experiment did not observe the influence on the apoptosis of pancreatic cancer cell.Conclusions:miR-223expression generally exists in all8pancreatic cancer cell lines observed in this experiment. Afterwards, dual luciferase reporter assay confirmed FOXOla as target of miR-223.Fianlly we demonstrated that miR-223facilitated pancreatic cancer cell proliferation, induction of re-distribution of cell cycle, but did not observe the influence on the apoptosis of pancreatic cancer cell.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Pancreatic tumors
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