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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
MiR-137 promotes anoikis through modulating the AKT signaling pathways in Pancreatic Cancer
Lin Li1,2,3,4, Zhiwei He1,2,3, Changhao Zhu1,2,3,4
1Guizhou Medical University, Guiyang, China.
Abstract:
Anoikis resistance is a fundamental feature of the survival of metastatic cancer cells during cancer progression. However, the mechanisms underlying anoikis resistance in pancreatic cancer (PC) are still unclear. MicroRNA-137 (miR-137) is a tumor suppressor that inhibits the proliferation and invasion of cancer cells through targeting multiple oncogenes. However, the effects and molecular mechanism of miR-137 on anoikis of PC are still unclear. Here we demonstrated that miR-137 was downregulated after the induction of anoikis model in time dependent. Function assays revealed that miR-137 promoted the pancreatic cancer cells anoikis in vitro and vivo. According to bioinformation analysis of clinical databases, we predicted that paxillin (PXN) was a target of miR-137. Further, TCGA analysis revealed that PXN was closely associated with the development of PC. Through loss-of-function studies, we demonstrated that PXN was a functional target of miR-137 on anoikis of PC cells. Moreover, we found that PXN promoted the activation of the AKT signaling pathways which was involving in the cancer cells anoikis. Together, our findings reveal that miR-137 plays a novel role during anoikis and may serve as a potential target for the detection and treatment of PC.
Insights
MicroRNA-137 (miR-137) promotes anoikis, or cell death, in pancreatic cancer cells. It targets paxillin (PXN), a protein that activates AKT signaling, suggesting miR-137 as a potential therapeutic target for pancreatic cancer.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Anoikis resistance is crucial for metastatic cancer cell survival.
- Mechanisms of anoikis resistance in pancreatic cancer (PC) remain largely unknown.
- MicroRNA-137 (miR-137) acts as a tumor suppressor, inhibiting cancer cell proliferation and invasion.
Purpose of the Study:
- To investigate the role and molecular mechanism of miR-137 in pancreatic cancer anoikis.
- To identify potential targets of miR-137 involved in anoikis regulation.
- To explore the therapeutic potential of miR-137 in pancreatic cancer.
Main Methods:
- Induction of anoikis models in pancreatic cancer cells (in vitro and in vivo).
- Bioinformatic analysis of clinical databases to predict miR-137 targets.
- Loss-of-function studies to validate target interactions and signaling pathways.
- TCGA analysis to assess the association of target genes with PC development.
Main Results:
- miR-137 expression was downregulated during anoikis induction in a time-dependent manner.
- miR-137 overexpression promoted anoikis in pancreatic cancer cells.
- Paxillin (PXN) was identified as a direct target of miR-137.
- PXN promotes pancreatic cancer cell anoikis by activating AKT signaling pathways.
Conclusions:
- miR-137 plays a novel role in promoting anoikis in pancreatic cancer.
- The miR-137/PXN/AKT axis is a key pathway regulating anoikis in PC.
- miR-137 represents a potential biomarker and therapeutic target for pancreatic cancer detection and treatment.
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