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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
Effects of miR-330 restoration on pancreatic cancer cells oncogenesis
Arezoo Farhadi1, Arash Poursheikhani2, Mohammad Foad Heidari3
1Department of Genetics and Molecular Medicine, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
Background:
Inappropriate expressions of various miRNAs have reported in different human malignancies. Evidence suggested that miR-330 may play as both onco-miR and/or tumor suppressor-miR in different cancers. In the present study, we evaluated effects of miR-330 on proliferation and migration of pancreatic cancer (PC) cells as well as underlying molecular mechanisms.
Design:
The expression of miR-330 was evaluated in clinical tissue samples of patients with PC. Transfection of the PC cells (PANC-1) by miR-330 was conducted by pCMV vector. The cancer-related genes expression was investigated in mRNA and protein level following transfection of the PC cells. Furthermore, the PC cells viability, invasion, migration, mitochondrial membrane potential, apoptosis, autophagy, and cell cycle profile were investigated after transfection by miR-330.
Results:
The results indicated that expression of miR-330 downregulated in patients with PC. Stable increase of miR-330 expression after transfection in PC cells reduces viability, mitochondrial membrane potential, invasion, and migration. Further assessments demonstrated that upregulation of miR-330 increases apoptosis and autophagy percentage in the PC cells. Moreover, a cell cycle arrest was observed in G1, Sub-G1, and S phases following transfection of the PC cells. These findings can be explained by modified mRNA and protein expression of apoptosis- and metastasis-related genes.
Conclusion:
Our study suggested that miR-330 acts as a tumor suppressor in PC cells, and revealed that upregulation of miR-330 may provide an effective therapeutic approach for overcoming progression and metastasis in patients with PC.
Insights
MicroRNA-330 (miR-330) is downregulated in pancreatic cancer (PC). Upregulating miR-330 suppresses PC cell proliferation and migration, offering a potential therapeutic strategy for PC treatment.
Area of Science:
- Molecular Biology
- Oncology
- Gene Regulation
Background:
- MicroRNAs (miRNAs) exhibit dysregulated expression in various human cancers.
- miR-330 has a dual role as an onco-miR or tumor suppressor-miR depending on the cancer type.
- Pancreatic cancer (PC) is a highly aggressive malignancy with limited therapeutic options.
Purpose of the Study:
- To investigate the role of miR-330 in pancreatic cancer (PC) cell proliferation and migration.
- To elucidate the underlying molecular mechanisms of miR-330 action in PC.
- To evaluate miR-330 as a potential therapeutic target for PC.
Main Methods:
- miR-330 expression analysis in clinical PC tissues.
- Transfection of PANC-1 PC cells with miR-330 using a pCMV vector.
- Assessment of cancer-related gene expression at mRNA and protein levels.
- Evaluation of cell viability, invasion, migration, mitochondrial membrane potential, apoptosis, autophagy, and cell cycle.
Main Results:
- miR-330 expression was found to be downregulated in PC tissues.
- Increased miR-330 expression in PC cells reduced viability, invasion, and migration.
- Upregulation of miR-330 enhanced apoptosis and autophagy.
- Transfection with miR-330 induced cell cycle arrest at G1, Sub-G1, and S phases.
- These effects correlated with altered expression of apoptosis- and metastasis-related genes.
Conclusions:
- miR-330 functions as a tumor suppressor in pancreatic cancer (PC).
- Upregulation of miR-330 demonstrates potential as a therapeutic strategy against PC progression and metastasis.
