New advances in technology have made it possible to further expand our knowledge in the scientific and medical field. Deshmukh, Arfuso, Newsholme, and Dhaarmarajan studies aims at using the Secreted Frizzled-Related Protein 4(sFRP4) to regulate the growth and explanation of cancer stem cells.
Treatment-resistant cancer stem cells are found in small number in Tumours.Evidence accrued overtime have suggested that tumours of several various tissue origins which includes the breast, prostate, and ovary holds a small sub-group of cells with stemlike capacity. It is often referred to as cancer stem cells or a tumour initiating cell which are usually self-renewal and migrant. Cancer stem cells also have the ability to flush out some of the toxic compounds chemicals and chemotherapeutic agents also known as cytotoxic agents as a result of their high expression of ATP dependent pump ABCG2, high DNA repair system, and the activation of survival cascades. In the introduction paragraph, the authors noted but did not cite some of the current researches they used in the study. Nonetheless, they indicated that several recent studies have managed to determine crucial signaling pathways and components that affect the differentiation and the stemness capacity of cancer stem cells. More studies was done that investigated the metabolic properties of Cancer stem cells based on their biochemical profile, mitochondrial respiratory activity, and glycolytic activity. The authors expanded on the Wnt signaling mentioned in the abstract. They recorded that Wnt signaling is fundamental in the development and maintenance of the normal tissues. In addition, the author explained the mechanism behind the Wnt signaling. It described that (sFrP4) being a Wnt antagonist it inhibits the canonical Wnt signalling when it binds to the Wnt frizzled and ligand receptors. The authors used specimen of cell lines from the prostate (PC3 and Lncap) and breast (MDA231 and MCF7) to isolate the cancer stem cells. The effect of sFRP4 was used on cancer stem cells isolated in culture medium with different levels of glucose content. They investigated the potential therapeutic effect of sFRP4 on cancer stem viability, the glucose uptake , the glutamine uptake, the glutamate secretion, and the NAD+/NADH ratio (1).
MATERIALS/METHODS
Different kinds of materials and methods were used in this research. The first one includes the Monolayer Cell Culture. The cells were “cultured in RPMI-1640 medium supplemented with 10%foetal bovine serum and 100 U/mL PenStrep. All the cells were maintained at 37 degrees celsius in a humid incubator with 5 % CO2” (13)
Cancer stem Cell Isolation was another alternative in this research. Cancer stem cells were cultured in serum-free medium containing different variables such as “DMEM-No Glucose, DMEM-Low Glucose, 5.5 mM , and DMEM-High Glucose, 25 mM supplemented with the growth factors bFGF (20 ng/mL) , EGF (20 ng/mL) , and 1× B27 and 100 U/mL PenStrep. CSC-enriched populations of cells were obtained by plating a single cell suspension of breast and prostate tumour cells at 10,000 cells/cm2 in SFM on Low-adherent six-well plates” (13).
Cancer Stem Cell Treatment was also used in this research. The cancer stem cells were treated in the study with purified sFRP4. A few other steps were followed as “the sensitization of Cancer Stem cell with SfRP4 was performed by adding SFRP4 to the cell culture at 250 pg/ mL for 24 hours at 37 degrees celsius in 5% CO2 incubator”(13).
Cell quantifying viability kits was used to determine the viable cells and cell surface markers that used the flow cytometry was used to enhance their identity (14).
A glucose bioluminescent uptake assay was applied to the cancer stem cell in addition to other protocols to measure the degree of the glucose uptake (14).
A luminometer was used to detect the Cancer stem cell extracellular metabolism and the qualitative NAD+/ NADH ratio (15).
Western blotting was used for the detection the metabolic gene-sets.
RESULTS
The first finding shows that the sFRP4 reduced the degree of the forming capacity of cancer stem cells irrespective of the glucose content.(Figure 1)
The second finding showed that the Cancer Stem Cell Viability is reduced by sFRP4 in Low and High Glucose Conditions.(Figure 2)
The third finding evaluates the coefficient correlation fo sFRP4 between the metabolic gene-set that comprised of AMPKB1 (AMP Kinase), mTOR (mammalian target of ramycin), GLUT1 (glucose transporter), SLC1A5 (glutamine transporter), BAD (Bcl-2), and PDHA1 (pyruvate dehydrogenase). The study indicated a negative correlation between the gene set and the sFRP4 vindicating that sFRP4 has an inhibiting effect on all the genes expect with BAD where it showed a positive correlation hence a pro-apoptotic capacity in both the tumours examined (Figure 3).
(Figure 4) The fourth objective was directed towards understanding the implication of SFRP4 and glucose on the uptake of glucose by the cancer stem cells. The results showed a higher glucose uptake by the breast cancer stem cells in the low glucose group contrary to the prostate cancer stem cells that recorded a high uptake of glucose in the groups with high glucose content. Besides, the level of the glucose uptake in all the CSCS significantly increased in the low glucose groups while decreased in the high glucose group hence indicating that sFRP4 directly affects the glycolytic flux translating to glycolytic activity (5).
The fifth objective that assessed the extracellular metabolic changes in extracellular metabolites shows that sFRP4 gravely inhibited the extracellular metabolite secretion.
The sixth objective that assessed the redox signature changes resting from the use of sFRP4 for treatment of CSCs showed that sFRP4 reduced the redox activity with an increase in the glucose concentration ( ).
The last objective assessing the impacts of sFRP4 of the metabolic capacity of the target protein of CSCs vindicated that the metabolic profile of the CSCs has varied changes (p. 49-50).
DISCUSSION/CONCLUSION
The authors discussed that for many decades it had been established that tumour cells actively depend on the glycolytic metabolism hence the alteration of the metabolic capacity of cancers proved a hallmark in the management of cancer. Many of the recent studies were noted to support the cancer stem cells glycolytic dependence theory hence the metabolic properties of cancer stem cells are key in understanding the tumours reproduction. Deshmukh, Arfuso, Newsholme. And Dharmarajan studies’ further demonstrated that sFRP4 has a consistent anti-proliferative impact, ability to cause spheroid disruptions and equally significantly inhibits the secretion of glutamate, uptake of glucose and glutamine, the redox signature and survival of cancer stem cells.
Basing on the study materials and methods, results and the discussion the authors concluded that sFRP4 plays an integral part in the treatment of prostate and breast cancer due to its ability to inhibit the prostate and breast Cancer stem cell metabolism. The finding is thus critical as more studies are directed towards management and treatment of cancer. Based on the study findings, it is concluded that sFRP4 significantly inhibited the survival mechanisms of the CSCs thus concluding that sFRP4 is key in the regulation growth of cancer stem cells and treatment of tumours.
In my opinion, this really gave me an insight in the process of how things are being discovered in regards to technology, science, and medicine. The author of this article along with his colleagues did tremendous amounts of precise research in order to see advance in Cancer stem cell field of study. Cancer Stem cell research is an important topic to discuss and research about. This topic is near and dear to my heart because my grandmother has recently been diagnosed with Stage III breast cancer. We are looking for other options other than chemotheraphy to help and provide her with the best possible cure out there. A well rounded published article goes a long way and inspires people to carry on and add on to the mystery of life through research.