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Published on: March 30, 2019
miR-155 Down Regulation by LNA Inhibitor can Reduce Cell Growth and Proliferation in PC12 Cell Line
Fatemeh Kouhkan1, Shaban Alizadeh, Saeid Kaviani
1Department of Genetics, Faculty of Basic Sciences, Tarbiat Modares University, Tehran, Iran.
Abstract:
MicroRNAs (miRNAs) are a class of small non coding regulatory RNAs that have key functions in multiple cell processes. Deregulation of these tiny miRNAs is involved in various human diseases. MiR-155 is one of the multifunctional miRNA that its over-expression has been found to be associated with different kinds of cancer such as leukemia, breast and colon cancers. It is thought that deregulation and over-expression of this microRNA may be associated with PC12 cell proliferation. So, the aim of this study was to investigate the role of miR-155 expression on PC12 cell growth. For this reason, PC12 cells were cultured and transfected by 3 different concentration (25, 50 and 75 nmol) of either LNA anti-miR-155 or scramble antisense in 24-well plate. Then, total RNA was extracted from transfected cells. miRNA cDNAs were synthesized from isolated total RNA. In the second step, miR-155 expression level was analyzed using the quantitative real-time polymerase chain reaction (QRT-PCR). MTT test was performed to evaluate cell viability. In the next step, apoptosis assay was assessed to investigate anti miR-155 effect on PC12 cells death. Obtained results were analyzed with t-test. MTT test revealed that cell viability of transfected cells with 75 nM of anti-miR- 155 to be reduced by half of the control and scramble groups (0.5 vs. 0.97 and 0.94). Our data suggest that miR-155 over-expression is associated with PC12 cell growth. So, miR-155 down regulation by anti-miR-155 could open up new ways to restrain brain tumor growth, as anti-miR-155 causes PC12 cells to repress.
Insights
MicroRNA-155 (miR-155) over-expression promotes PC12 cell growth. Inhibiting miR-155 with anti-miR-155 significantly reduces cell viability, suggesting a therapeutic target for brain tumors.
Area of Science:
- Molecular Biology
- Cancer Research
- Cell Biology
Background:
- MicroRNAs (miRNAs) are small non-coding RNAs regulating gene expression.
- Dysregulated miRNA expression is implicated in various human diseases, including cancers.
- MiR-155 over-expression is linked to several cancers, and its role in PC12 cell proliferation is investigated.
Purpose of the Study:
- To investigate the role of miR-155 expression in PC12 cell growth.
- To determine the effect of miR-155 down-regulation on PC12 cell viability and apoptosis.
Main Methods:
- PC12 cells were transfected with varying concentrations of LNA anti-miR-155 or scramble antisense.
- Quantitative real-time PCR (QRT-PCR) was used to analyze miR-155 expression levels.
- MTT assays and apoptosis assays were performed to assess cell viability and cell death.
Main Results:
- Transfection with 75 nM anti-miR-155 reduced PC12 cell viability by approximately 50% compared to control and scramble groups.
- Down-regulation of miR-155 using anti-miR-155 led to decreased PC12 cell proliferation.
- Apoptosis assays indicated that anti-miR-155 treatment induced cell death in PC12 cells.
Conclusions:
- MiR-155 over-expression is associated with increased PC12 cell growth.
- Inhibiting miR-155 with anti-miR-155 demonstrates potential for restraining PC12 cell proliferation.
- Targeting miR-155 may offer a novel therapeutic strategy for brain tumor treatment.
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