Multiple SNPs Downregulate Gene Expression of Matrix Metallopeptidase 2 in MCF7 Breast Cancer Cells

@#Introduction: On a global scale, breast cancer contributes the highest cancer-related deaths in women due to metastasis which renders the treatments ineffective and non-targeted. The members of Matrix Metallopeptidases, particularly Matrix Metallopeptidase 2 (MMP2), are among the key players in breast cancer metastasis. In most cases, MMP2 was markedly upregulated and linked to poor prognosis. In a previous study, in silico analyses revealed that several coding single nucleotide polymorphisms (SNPs) of MMP2 were shown to reduce gene expression and mRNA stability of MMP2 in Malaysian breast cancer patients. Therefore, to validate the in silico predictions, the objective of this study was to determine the effects of multiple coding SNPs of MMP2 on the gene expression and mRNA stability of MMP2 in breast cancer cells. Methods: In the current study, breast adenocarcinoma MCF7 cells were transfected with MMP2 wild type and variant containing the coding SNPs. After confirmation of transfection by DNA sequencing, the gene expression level of MMP2 was evaluated by quantitative reverse transcription polymerase chain reaction (RT-qPCR) whereas mRNA stability of MMP2 was determined following treatment with actinomycin D. Results: MMP2 wild type and variant were successfully transfected in MCF7 cells based on sequencing and PCR analysis. It was found that the presence of coding SNPs lowered the gene expression level of MMP2, but not the stability of MMP2 mRNA. Conclusion: This study supports the in silico effects of MMP2 coding SNPs on its gene expression in an in vitro model.

The Potentiality of Citral in Targeting Breast Cancer Multicellular Tumour Spheroids (MTS)

@#Introduction: As the high incidence of breast cancer has a profound impact on a global scale, there is a critical need to improve the clinical outcome of the patients, including efforts to utilize bioactive natural products as treatment or preventive measures. Citral, the essential oil of lemongrass has been reported to possess cytotoxicity in breast cancer cell line . The aim of present study was to determine the capability of citral in targeting aldehyde dehydrogenase-positive (ALDH+) cells in breast cancer cells. Methods: Both MCF-7 and MDA-MB-231 cells were cultured in serum-free media to generate multicellular tumour spheroids for the evaluation of citral as an antiproliferative agent. The cells were treated with identified IC50 (50±4.30 µM and 56±3.17 µM of citral, respectively) to investigate the cytotoxicity of citral. Staining using Propidium Iodide (PI) and Hoechst 33342 was carried out to determine cell proliferation and viability. Finally, ALDH+ cells were quantified via ALDEFLUOR assay. Analysis of differences was carried out by analysis of variance (ANOVA) and independent t-test with p<0.05 considered statistically significant. Results: The size of spheroids in both cancer cell lines were reduced after treatment with the citral. PI and Hoechst 33342 staining also revealed that citral gave rise to a mixture of cells that are normal and undergoing apoptosis and necrosis. ALDEFLUOR assay analysis revealed citral significantly (p <0.05 ) inhibited the population of ALDH+ cells in MCF7 cells. Conclusion: It was demonstrated that citral reduced the ALDH+ cell population in MCF7 breast cancer spheroids by inhibiting the ALDH activity.

Matrix Metallopeptidase 3 Polymorphisms: Emerging genetic Markers in Human Breast Cancer Metastasis / 한국유방암학회지

Matrix metallopeptidase 3 or MMP3, is a zinc-dependent proteolytic enzyme that is involved in various physiological processes via modification of the extracellular matrix. In particular, its over-expression has been associated with cancer metastasis and tumor growth in various cancers including breast cancer. MMP3 gene expression is regulated by several factors such as DNA polymorphisms which also serve as risk factors for breast cancer. As such, DNA polymorphisms of MMP3 have the potential to be utilized as genetic biomarkers for prediction and prognosis of metastatic breast cancer. Presently, genome-wide association studies of MMP3 gene polymorphisms which are associated with breast cancer risk and patient survival in a variety of populations are reviewed. In order to understand the potential role of MMP3 polymorphisms as genetic markers for breast cancer metastasis, the domain structure of MMP3, the regulation of its expression and its role in breast cancer metastasis are also briefly discussed in this review. The emergence of MMP3 gene polymorphisms as prognostic biomarker candidates for breast cancer metastasis may contribute towards improving targeted therapies and categorization of breast cancer cases in order to provide a better and more accurate prognosis.