Title |
MicroRNA-29b attenuates non-small cell lung cancer metastasis by targeting matrix metalloproteinase 2 and PTEN
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Published in |
Journal of Experimental & Clinical Cancer Research, June 2015
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DOI | 10.1186/s13046-015-0169-y |
Pubmed ID | |
Authors |
Hongyan Wang, Xiaoying Guan, Yongsheng Tu, Shaoqiu Zheng, Jie Long, Shuhua Li, Cuiling Qi, Xiaobin Xie, Huiqiu Zhang, Yajie Zhang |
Abstract |
Our pilot study using miRNA PCR array found that miRNA-29b (miR-29b) is differentially expressed in primary cultured CD133-positive A549 cells compared with CD133-negative A549 cells. Ten human non-small cell lung cancer (NSCLC) cell lines and samples from thirty patients with NSCLC were analyzed for the expression of miR-29b by quantitative RT-PCR. Bioinformatics analysis combined with tumor metastasis PCR array showed the potential target genes for miR-29b. miR-29b lentivirus and inhibitors were transfected into NSCLC cells to investigate its role on regulating cell proliferation which was measured by CCK-8 assay in vitro and nude mice xenograft tumor assay in vivo. Cell motility ability was evaluated by transwell assay. The target genes of miR-29b were determined by luciferase assay, quantitative RT-PCR and western blot. Bioinformatics analysis combined with tumor metastasis PCR array showed that matrix metalloproteinase 2 (MMP2) and PTEN could be important target genes of miR-29b. The expression of miR-29b was down regulated in NSCLC tissues compared to the normal tissues. Clinicopathological analysis demonstrated that miR-29b had significant negative correlation with lymphatic metastasis. The gain-of-function studies revealed that ectopic expression of miR-29b decreased cell proliferation, migration and invasion abilities of NSCLC cells. In contrasts, loss-of-function studies showed that inhibition of miR-29b promoted cell proliferation, migration and invasion of NSCLC cells in vitro. Nude mice xenograft tumor assay confirmed that miR-29b inhibited lung cancer growth in vivo. High-invasion (A549-H) and low-invasion (A549-L) NSCLC cell sublines from A549 cells were created by using the repeated transwell assay aimed to confirm the effect of miR-29b on migration and invasion of NSCLC. Furthermore, the dual-luciferase reporter assay demonstrated that miR-29b inhibited the expression of the luciferase gene containing the 3'-UTRs of MMP2 and PTEN mRNA. Western blotting and quantitative RT-PCR indicated that miR-29b down-regulated the expression of MMP2 at the protein and mRNA levels. Taken together, our results demonstrate that miR-29b serves as a tumor metastasis suppressor, which suppresses NSCLC cell metastasis by directly inhibiting MMP2 expression. The results show that miR-29b may be a novel therapeutic candidate target to slow NSCLC metastasis. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 43 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Master | 10 | 23% |
Student > Ph. D. Student | 8 | 19% |
Student > Bachelor | 4 | 9% |
Researcher | 4 | 9% |
Unspecified | 2 | 5% |
Other | 7 | 16% |
Unknown | 8 | 19% |
Readers by discipline | Count | As % |
---|---|---|
Medicine and Dentistry | 9 | 21% |
Agricultural and Biological Sciences | 9 | 21% |
Biochemistry, Genetics and Molecular Biology | 7 | 16% |
Pharmacology, Toxicology and Pharmaceutical Science | 3 | 7% |
Unspecified | 2 | 5% |
Other | 6 | 14% |
Unknown | 7 | 16% |