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An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
MicroRNA‑506 participates in pancreatic cancer pathogenesis by targeting PIM3
Jundong Du1, Xi Zheng2, Shouwang Cai3
1Department of Hepatobiliary Surgery, The First Affiliated Hospital to the General Hospital of the PLA, Beijing 100853, P.R. China.
Abstract:
MicroRNA (miRNA) is a type of short non-coding RNA that suppresses the expression of protein coding genes by partial complementary binding to the 3'‑untranslated regions (UTRs) of mRNAs. miRNA expression alterations are involved in the initiation, progression and metastasis of human cancer and it has been suggested that miRNAs function as tumor suppressors as well as oncogenes in cancer development. PIM-3 is a member of the proto-oncogene PIM family, the aberrant expression of which exists in human pancreatic cancer tissues. There are reports indicating that overexpression of PIM3 is associated with the promotion of pancreatic cancer cell proliferation. The aim of the present study was to identify micro (mi)RNAs that regulate the expression of the oncogene PIM3 in PC. It was confirmed that the expression of PIM3 was regulated by miRNAs through an AGO2 knockout experiment. Subsequently, a dual luciferase assay system was constructed and used to screen 13 selected miRNAs that may target the PIM3 3'UTR directly. The results indicated that miR‑15a/b, miR‑16, miR‑33a/b, miR‑124, miR‑195 and miR‑506 repressed the luciferase activity by targeting the PIM3 3'UTR. However, only the expression of miR‑506 was negatively correlated with PIM3 expression in PC tissues (r=‑0.38, P=0.017). Furthermore, a biological functional study indicated that miR‑506 functioned as a tumor suppressor by repressing PC cell proliferation, which was partially reversed by PIM3 overexpression. To the best of our knowledge, the present study was the first to reveal the tumor suppressor function of miR‑506 in PC, which has the potential to be employed in the diagnosis and treatment of PC.
Insights
MicroRNA-506 (miR-506) acts as a tumor suppressor by inhibiting pancreatic cancer (PC) cell proliferation. This study identifies miR-506 as a regulator of the oncogene PIM3, offering potential for PC diagnosis and treatment.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- MicroRNAs (miRNAs) are key regulators of gene expression implicated in human cancer.
- Aberrant expression of the proto-oncogene PIM3 is observed in pancreatic cancer (PC) and promotes cell proliferation.
- Understanding miRNA regulation of PIM3 is crucial for PC research.
Purpose of the Study:
- To identify specific microRNAs (miRNAs) that regulate the expression of the oncogene PIM3 in pancreatic cancer (PC).
- To investigate the functional role of identified miRNAs in PC development and progression.
Main Methods:
- Utilized an AGO2 knockout experiment to confirm miRNA-mediated regulation of PIM3.
- Employed a dual luciferase assay system to screen for miRNAs targeting the PIM3 3'UTR.
- Correlated miRNA and PIM3 expression in PC tissues and performed functional studies.
Main Results:
- Several miRNAs, including miR-506, were found to directly target the PIM3 3'UTR.
- miR-506 expression showed a negative correlation with PIM3 expression in PC tissues.
- miR-506 demonstrated tumor suppressor activity by inhibiting PC cell proliferation, partially reversed by PIM3 overexpression.
Conclusions:
- miR-506 functions as a tumor suppressor in pancreatic cancer by repressing PIM3 expression and inhibiting cell proliferation.
- This study is the first to reveal the tumor suppressor role of miR-506 in PC.
- miR-506 holds potential for diagnostic and therapeutic applications in pancreatic cancer.
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