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Published on: August 23, 2019
MicroRNA-145 replacement effect on growth and migration inhibition in lung cancer cell line
Navaz Sadeghiyeh1, Nasser Sehati1, Behzad Mansoori2
1Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Background:
Lung cancer is the main cause of cancer death in males and females worldwide. Reduced expression of miR-145 has been reported in many types of cancers. In this study, we transfected miR-145 into lung cancer cells by vector-based miR-145, and investigated the effects of this intervention on growth and migration inhibition of cancer cells as well on the expression of targeted genes.
Methods:
IC50 of Geneticin (G418) antibiotic was measured using MTT test in NSCLC cell lines. miR-145 was transfected into lung cancer cells by jetPEI. qRT-PCR was used to evaluate the transcript level of the miR-145 and expression for KRAS, MMP-9, vimentin, caspase-3, caspase-8 and caspase-9 genes in A549 cells. MTT assay was used to evaluate the proliferation inhibition of cancer cells. Wound healing assay was used to check the migration status of transfected lung cancer cells. The apoptosis induction was assessed by DAPI staining assay.
Results:
The MTT assay showed that the IC50 of Genticin was 494.1 μg/ml. The results of the qRT-PCR showed increased expression level of miR-145 and downregulation of KRAS, MMP-9, and vimentin expression in A549 transfected cells compared with the control group. The MTT assay results demonstrated inhibition of cancer cell proliferation after miR-145 replacement. Wound healing assay results revealed that migration was reduced upon miR-145 transfection. The transfected cell displayed increased apoptosis rate by inducing caspase-3 and caspase-9 mRNA expression.
Conclusion:
The results of this study showed that increased miR-145 expression exerted a critical role in subsiding the growth, survival, and migration of lung cancer cell line.
Insights
Restoring miR-145 in lung cancer cells significantly inhibits cancer growth and migration. This microRNA replacement therapy downregulates key oncogenes like KRAS and MMP-9, promoting apoptosis and reducing survival.
Area of Science:
- Molecular Biology
- Oncology
- Gene Expression Regulation
Background:
- Lung cancer remains a leading cause of cancer mortality globally.
- Reduced microRNA-145 (miR-145) expression is implicated in various cancers.
- Investigating miR-145's role in lung cancer pathogenesis is crucial.
Purpose of the Study:
- To investigate the therapeutic potential of restoring miR-145 expression in lung cancer cells.
- To evaluate the impact of miR-145 on cancer cell proliferation, migration, and apoptosis.
- To analyze the effect of miR-145 on the expression of target genes including KRAS, MMP-9, and vimentin.
Main Methods:
- Non-small cell lung cancer (NSCLC) cell lines were used.
- miR-145 was introduced into lung cancer cells via vector-based transfection.
- Quantitative real-time PCR (qRT-PCR) assessed gene expression levels.
- MTT assays measured cell proliferation and drug sensitivity.
- Wound healing and DAPI staining assays evaluated cell migration and apoptosis, respectively.
Main Results:
- miR-145 restoration led to increased miR-145 levels and decreased expression of KRAS, MMP-9, and vimentin.
- Transfection with miR-145 significantly inhibited lung cancer cell proliferation and migration.
- Apoptosis was induced, evidenced by increased caspase-3 and caspase-9 mRNA expression.
Conclusions:
- Increased miR-145 expression plays a vital role in suppressing lung cancer cell growth, survival, and migration.
- miR-145 holds potential as a therapeutic agent for lung cancer.
- Targeting miR-145 could be a promising strategy in lung cancer treatment.
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