Dysregulation of miR-15a and miR-214 in human pancreatic cancer

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Abstract

Background: Recent reports indicate that microRNAs (miRNAs) play a critical role in malignancies. However, the role that miRNAs play in pancreatic cancer remains to be determined. The purpose of this study was to investigate aberrantly expressed miRNAs in pancreatic cancer tissues and demonstrate their roles in disease progression. Results. We detected the expression patterns of miRNAs in 10 pancreatic cancer tissues and their adjacent benign tissues by quantitative real time-PCR (qRT-PCR) and found that miR-15a and miR-214 were dysregulated in the tumor samples. This is the first time that miR-214 has been identified as aberrantly expressed in pancreatic cancer. In vitro experiments showed that overexpression of miR-15a inhibited the viability of pancreatic cancer cells, whereas overexpression of miR-214 decreased the sensitivity of the cells to gemcitabine (GEM). Furthermore, we identified WNT3A and FGF7 as potential targets of miR-15a and ING4 as a target of miR-214. Conclusions. Aberrant expression of miRNAs such as miR-15a and miR-214 results in different cellular effects in pancreatic cancer. Downregulation of miR-15a might contribute to proliferation of pancreatic cancer cells, whereas upregulation of miR-214 in pancreatic cancer specimens might be related to the poor response of pancreatic cancer cells to chemotherapy. MiR-15a directly targets multiple genes relevant in pancreatic cancer, suggesting that it may serve as a novel therapeutic target for treatment of the disease. © 2010 Zhang et al; licensee BioMed Central Ltd.

Figures

  • Table 1 Expression of miRNAs in pancreatic cancer specimens compared with adjacent benign pancreatic tissues
  • Figure 1 Expression patterns of miR-15a and miR-214. qRT-PCR was performed to detect (A) miR-214 and (B) miR-15a expression in 10 pancreatic cancer tissues and their adjacent benign pancreatic tissues. Expression levels of miRNAs in adjacent benign pancreatic tissues were set as 1. Relative values were calculated to indicate the frequency of miRNA expression downregulated or upregulated in pancreatic cancer.
  • Figure 2 MiR-15a and miR-214 have different roles in pancreatic cancer cells. (A) qRT-PCR was used to investigate miRNA transfection efficiency. Both miR-15a and miR-214 were significantly increased compared to their mimics-NC (control) in BxCP-3 cells. (B) The viability of BxCP-3 cells after transfection was measured by CCK-8 assay. (C) Cell viability was measured using the CCK-8 assay in BxCP-3 cells treated with 10 μM GEM at 24, 48 and 72 hrs. (D) The expression pattern of miR-214 was detected by qRT-PCR in BxCP-3 cells treated with GEM. (E) The CCK8 assay was used to measure the inhibition effect of miR-214 on apoptosis of BxCP-3 cells induced by GEM. BxCP-3 cells were transfected with H2O (MOCK), mimics-NC (NC), and miR-214 mimics (miR-214). Significant differences (* p < 0.05; ** p < 0.01) compared with the control were calculated using Dunnett’s test or the paired Student’s t-test.
  • Table 2 Target validation for miR-15a
  • Table 3 Target validation for miR-214
  • Figure 3 Target validation of miR-15a and miR-214. (A) The 3’-UTR of WNT3A and FGF7 contain predicted MREs for miR-15a. (B) The 3’-UTR of ING4 contains the predicted MRE for miR-214. (C) A luciferase assay was used to measure the activity of the 3’-UTR reporter in 293T cells. MiR-15a inhibited the activity of WNT3A and FGF7 3’-UTR reporters, whereas miR-214 inhibited the activity of the ING4 3’-UTR reporter.
  • Table 4 qRT-PCR Primers for miRNAs and U6

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APA

Zhang, X. J., Ye, H., Zeng, C. W., He, B., Zhang, H., & Chen, Y. Q. (2010). Dysregulation of miR-15a and miR-214 in human pancreatic cancer. Journal of Hematology and Oncology, 3. https://doi.org/10.1186/1756-8722-3-46

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