Title:The Effect of Silencing MiR-4270 on Apoptosis in HCC Cell Line
Volume: 19
Issue: 1
Author(s): Hanieh Gholamia, Hassan Akrami*, Hosseinali Sassan, Nasrollah Erfani, Mohammad Reza Fattahi and Mojdeh Heidari
Affiliation:
- Gastroenterohepatology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
Keywords:
Hepatocellular carcinoma, apoptosis, hsa-miR4270, HepG2 cell line, BAX, BCL2, Caspase 3.
Abstract:
Background: Hepatocellular carcinoma (HCC) is the most common type of cancer.
Although HCC treatment has greatly improved over the past few decades, patient survival rates
are still very low. Therefore, it is essential to find new treatments for HCC. Apoptosis has been
shown to be the most effective in disrupting cancer growth. Improper functioning of proteins in
apoptosis can lead to cancer growth. MicroRNAs (miRNAs) are key regulators in the development
and progression of HCC.
Objective: Irregular expression of miRNAs involved in apoptosis signaling can lead to tumorigenesis.
Therefore, we investigated the effect of the hsa-miR-4270 inhibitor on cell proliferation
and apoptosis.
Methods: HepG2 cells were cultured at 37°C and 95% air. Transfection of HepG2 cells was
performed by miR-4270 inhibitor and lipofectamine 2000. Cell proliferation of HepG2 cells was
determined with MTT assay and different concentrations of miR-4270 specific inhibitors. DNA
laddering assay was performed to evaluate the induction of apoptosis. Finally, the transcription
level of genes involved in apoptosis, including BAX, BCL2, Caspase3, and p53, was measured
by real-time RT-PCR.
Results: The results of MTT and DNA laddering assays showed that the miR-4270 inhibitor
declined cell proliferation and induced apoptosis in HepG2 cells. Also, the results of quantitative
real-time RT-PCR indicated an upregulation of transcription of BAX, p53 and Caspase3 genes
and a decline in expression of BCL2 gene.
Conclusion: Taken together, we found hsa-miR-4270 inhibitor decreased cell proliferation and
induced apoptosis in the HepG2 cell line, which can be used as a new therapeutic strategy for
HCC patients.
