Knockdown of CD-74 in the Proliferative and Apoptotic Activity of Breast Cancer Cells.

BACKGROUND:The cluster of differentiation (CD) 74 is known for its immunological functions and its elevated level was reported in various cancer cells. AIM:The aim of the present study was to investigate the expression and potential roles of CD74 in the proliferative and apoptotic activity of breast cancer. METHODS:Expression of CD74, macrophage migration inhibitory factor (MIF) and CD44 was assayed in CAMA-1 and MDA-MB-231 cell lines using flow cytometry. CD74 was knocked down using CD74 siRNA-transfection in CAMA-1, and MDA-MB-231 cells and proliferation and apoptosis were determined in the transfected breast cancer cells. RESULTS:The data showed that CD74, MIF and CD44 were expressed in breast cancer cell lines and were associated with cell proliferation and apoptosis. Correlation analysis revealed that CD74 was positively correlated and colocalised with MIF on the cell-surface of CAMA-1 and MDA-MB-231. The knockdown of CD74 significantly reduced CAMA-1 and MDA-MB-231 cell proliferation and increased the level of apoptotic cells. CONCLUSION:We concluded that the interactions of CD74 with MIF and CD74 with CD44 could be a potential tumour marker for breast cancer cells. Moreover, the level of co-expression of MIF and CD74 or CD44 could be a surrogate marker for the efficacy of anti-angiogenic drugs, particularly in breast cancer tumours. In short, the study revealed the potential roles of CD74 in the proliferation and apoptosis of breast cancer which may serve as a potential therapeutic target for breast cancer

The impact of FXR-activation on the proliferative and invasive potential of breast cancer cell lines.

Advanced breast carcinoma is a main cause of mortality affecting women. It is associated with poor prognosis; therefore, understanding the molecular mechanisms of invasive malignancies is critical in order to discover new therapies and to optimise current therapies. Degradation of the Extracellular Matrix (ECM) is a crucial step in tumour growth and invasion, allowing the cancerous cells to metastasize to distant organs and form secondary tumours. Matrix metalloproteinases (MMP) are a family of zinc-dependant endopeptidases that have the capability to degrade the ECM, enabling tumours cells to invade and migrate. Hence, MMP overexpression and/or enhanced activity are positively associated with breast cancer metastasis and invasion. Two MMPs have been shown to be involved in breast cancers, gelatinase A (MMP-2) and gelatinase B (MMP-9). The activity of MMPs is tightly regulated by endogenous inhibitors that are called tissue inhibitors of metalloproteinases (TIMPs). There are four different endogenous negative regulator proteins of MMPs, with TIMP-1 and TIMP-2 inhibiting MMP-9 and -2, respectively. Due to their potential role in tumour metastasis, a great interest has grown in developing novel methods to inhibit the MMPs as there is a direct relation between the expression of MMPs and the invasiveness of the tumours. The Farnesoid X Receptor (FXR) is a nuclear receptor that is highly expressed in breast cancers, and has been reported to be involved in in regulation of MMP and TIMP activity in hepatic and vascular tissues. The rationale of the current study was to investigate whether FXR is a novel regulator of matrix metalloprotease-2 and -9 in metastatic breast cancer cells, and hence may represent a novel therapeutic target. Two FXR agonists were used to measure their effects on breast cancer cells MDA-MB-231, MDA-MB-468 (triple negative), MCF-7 (Estrogen receptor positive) and normal cells MCF10A: chenodeoxycholic acid (CDCA) is an endogenous (low-affinity, low-selectivity) ligand, while 3-[2-[2-Chloro-4-[[3-(2, 6-dichlorophenyl)-5-(1-methylethyl)-4isoxazolyl]methoxy]phenyl]ethenyl]benzoic acid (GW4064) is a synthetic (high affinity, high-selectivity) ligand. Cell viability, protein and mRNA levels of matrix metalloprotease -2 and -9 and TIMP-2 and -1, and cell migration were assessed using both molecular and cellular techniques. Both FXR agonists decreased breast cancer and normal breast cell viability, with the effects more significant in the triple negative cells, suggesting a potential targeting towards aggressive, hard-to-treat cancer cell-types. However, the FXR ligands didn’t alter mRNA and protein levels of MMP-2, MMP-9, TIMP-1 and TIMP-2 intracellularlly or extracellularlly, suggesting that this cytotoxic effect may not be via MMP. FXR ligands also had no effect on breast cancer cell migration, which is consistent with the suggestion that FXR activation has a general cytotoxic effect on tumour cells, but does not directly impact on tumour migration. In conclusion, FXR activation does not impact on markers of breast cancer metastasis, suggesting that its potential as a therapeutic target to prevent tumour progression may be limited. However, the cytotoxic effect on cancer cells is promising, suggesting that these agonists may still possess some potential for breast cancer therapy

The impact of FXR-activation on the proliferative and invasive potential of breast cancer cell lines.

Advanced breast carcinoma is a main cause of mortality affecting women. It is associated with poor prognosis; therefore, understanding the molecular mechanisms of invasive malignancies is critical in order to discover new therapies and to optimise current therapies. Degradation of the Extracellular Matrix (ECM) is a crucial step in tumour growth and invasion, allowing the cancerous cells to metastasize to distant organs and form secondary tumours. Matrix metalloproteinases (MMP) are a family of zinc-dependant endopeptidases that have the capability to degrade the ECM, enabling tumours cells to invade and migrate. Hence, MMP overexpression and/or enhanced activity are positively associated with breast cancer metastasis and invasion. Two MMPs have been shown to be involved in breast cancers, gelatinase A (MMP-2) and gelatinase B (MMP-9). The activity of MMPs is tightly regulated by endogenous inhibitors that are called tissue inhibitors of metalloproteinases (TIMPs). There are four different endogenous negative regulator proteins of MMPs, with TIMP-1 and TIMP-2 inhibiting MMP-9 and -2, respectively. Due to their potential role in tumour metastasis, a great interest has grown in developing novel methods to inhibit the MMPs as there is a direct relation between the expression of MMPs and the invasiveness of the tumours. The Farnesoid X Receptor (FXR) is a nuclear receptor that is highly expressed in breast cancers, and has been reported to be involved in in regulation of MMP and TIMP activity in hepatic and vascular tissues. The rationale of the current study was to investigate whether FXR is a novel regulator of matrix metalloprotease-2 and -9 in metastatic breast cancer cells, and hence may represent a novel therapeutic target. Two FXR agonists were used to measure their effects on breast cancer cells MDA-MB-231, MDA-MB-468 (triple negative), MCF-7 (Estrogen receptor positive) and normal cells MCF10A: chenodeoxycholic acid (CDCA) is an endogenous (low-affinity, low-selectivity) ligand, while 3-[2-[2-Chloro-4-[[3-(2, 6-dichlorophenyl)-5-(1-methylethyl)-4isoxazolyl]methoxy]phenyl]ethenyl]benzoic acid (GW4064) is a synthetic (high affinity, high-selectivity) ligand. Cell viability, protein and mRNA levels of matrix metalloprotease -2 and -9 and TIMP-2 and -1, and cell migration were assessed using both molecular and cellular techniques. Both FXR agonists decreased breast cancer and normal breast cell viability, with the effects more significant in the triple negative cells, suggesting a potential targeting towards aggressive, hard-to-treat cancer cell-types. However, the FXR ligands didn’t alter mRNA and protein levels of MMP-2, MMP-9, TIMP-1 and TIMP-2 intracellularlly or extracellularlly, suggesting that this cytotoxic effect may not be via MMP. FXR ligands also had no effect on breast cancer cell migration, which is consistent with the suggestion that FXR activation has a general cytotoxic effect on tumour cells, but does not directly impact on tumour migration. In conclusion, FXR activation does not impact on markers of breast cancer metastasis, suggesting that its potential as a therapeutic target to prevent tumour progression may be limited. However, the cytotoxic effect on cancer cells is promising, suggesting that these agonists may still possess some potential for breast cancer therapy

Therapeutic Properties of Vanadium Complexes

Vanadium is a hard, silver-grey transition metal found in at least 60 minerals and fossil fuel deposits. Its oxide and other vanadium salts are toxic to humans, but the toxic effects depend on the vanadium form, dose, exposure duration, and route of intoxication. Vanadium is used by some life forms as an active center in enzymes, such as the vanadium bromoperoxidase of ocean algae and nitrogenases of bacteria. The structure and biochemistry of vanadate resemble those of phosphate, hence vanadate can be regarded as a phosphate competitor in a variety of biochemical enzymes such as kinases and phosphatases. In this review, we describe the biochemical pathways regulated by vanadium compounds and their potential therapeutic benefits for a range of disorders including type 2 diabetes, cancer, cardiovascular disease, and microbial pathology