ABSTRACT
@#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.
ABSTRACT
Azadirachta indica [Neem] has been used traditionally for many centuries. Some impressive therapeutic qualities have been discovered. However, the therapeutic effect of neem leaf extract in 4T1 breast cancer has not been documented. The purpose of the present study is to investigate the therapeutic effect of ethanolic Neem leaf extract in an in vivo 4T1 breast cancer model in mice. A total of 84 female BALB/c mice were divided randomly into 7 groups [3 non-cancerous groups and 4 cancerous groups] consisting of 12 mice per group. The 3 non-cancerous groups were normal mice treated with 0.5% of Tween 20 in phosphate buffer saline [PBS] [NC], 250 mg/kg Neem [N250] or 500 mg/kg Neem [N500]. The 4 cancerous groups were; cancer controls treated with 0.5% of Tween 20 in PBS [CC], and cancerous mice treated with 0.5 micro g/mL tamoxifen citrate [CT], 250 mg/kg Neem leaf extract [CN 250] or 500 mg/kg Neem leaf extract [CN 500]. Terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL] assays were used to evaluate apoptosis [cell death] in the breast cancer tissues. SPSS software, version 14 was used for statistical analysis. Statistical significance was defined as p<0.05.Non parametric analysis of variance [ANOVA] was performed with the Kruskal Wallis test for the TUNEL assays. Parametric data among the groups was compared using ANOVA. TUNEL assays showed that the CN 250 and CN 500 groups had a higher incidence of apoptosis compared with the cancer controls. The findings showed that neem leaf extract induces apoptosis in 4T1 breast cancer BALB/c mice