Inhibitory effect of anti-miR-196a on invasion and migration of human pancreatic cancer PANC-1 cells |
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Authors: | ZHANG Shi-neng ZHUANG Xiao-hong TANG Jian ZHUANG Yan-yan HUANG Feng-ting CHENG Wen-bo CHEN Wen-jie |
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Institution: | 1. Department of Gastroenterology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China;2. Department of Oncology and Hematology, Hainan Provincial Nongken Hospital, Haikou 571103, China |
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Abstract: | AIM:To investigate the expression of miR-196a in different pancreatic cancer cell lines and to observe the effect of anti-miR-196a on the biological behaviors of human pancreatic cancer PANC-1 cells. METHODS:The expression of miR-196a in the pancreatic cancer cells was examined by real-time quantitative PCR. Anti-miR-196a was chemically synthesized and transfected into PANC-1 cells by Lipofectamine 2000. The cell proliferation was measured by CCK-8 assay. The apoptosis was determined by flow cytometry. Matrigel invasion assay was performed to examine the migration and invasion of the tumor cells. The wild-type and mutant-type NF-κB inhibitor α(NFKBIA) 3'UTR luciferase reporter vectors were constructed. The relative activity of Renilla luciferase was detected to confirm the binding site of miR-196a on NFKBIA mRNA. RESULTS:The expression of miR-196a in human pancreatic cancer cell lines was significantly higher than that in human pancreatic ductal epithelial H6c7 cells. The expression of anti-miR-196a in miR-196a group was down-regulated. After transfected with miR-196a, no change of cell proliferation and apoptosis was observed, but the abilities of invasion and migration were reduced(P<0.01). Compared with negative control, wild-type NFKBIA3'UTR or mutant-type NFKBIA 3'UTR, cotransfection of anti-miR-196a and wild-type NFKBIA 3'UTR significantly increased the relative activity of Renilla luciferase. CONCLUSION:miR-196a is one of the oncomiRs and may be a target microRNA of human pancreatic cancer for gene therapy. |
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Keywords: | Pancreatic neoplasms MicroRNA Neoplasm invasiveness |
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