Introduction
What is cancer? Cancer can be grouped as more than 100 different diseases and develops all over the body [1]. Abnormal cells will divide uncontrollably leading to the destruction of body tissues. Genetic changes affect the body’s logical processes. The cells that grow uncontrollably may form a tumor that can be benign or cancerous. A cancerous tumor can grow and sometimes it spreads to other parts of the body. A benign tumor can grow, but it will not spread to other parts of the body. Some cancer types don’t form a tumor. So why has cancer not been cured, when billions of Rands have been spent on research over the years? It is because cancer is very complex, not just one disease and one cure a one-size-fits-all cure is not possible. Cancer cells evade the immune system or suppress the normal immune response. A chemical compound can be added to the DNA of a cancer cell and suppress the activity of specific genes, causing cancer to hide in plain sight without causing an immune response. Ingestion of kombucha fermented tea has been associated with many health benefits because there are claims that it is a prophylactic agent, but it has not been scientifically proven. The beneficial effects of kombucha tea are that the bioactive compounds act synergistically, meaning working together, to accomplish more than they could alone. Kombucha is a culture consisting of a symbiosis between acetic acid bacteria (e.g. Gluconobacter oxydans) and different yeasts (e.g. Saccharomyces sp.) and some lactic acid bacteria have also been isolated from some symbiotic kombucha culture. Kombucha tea is a fermented drink consisting of a mixture of the kombucha culture (bacteria and yeast), green and/or black tea, and sugar. Kombucha tea tastes slightly acidic and slightly carbonated. Based on personal observations and testimonials kombucha tea is safe for human consumption. Studies are analyzed to experimentally confirm the health benefits of kombucha tea.
Background
As a cancerous tumor grows, the bloodstream or lymphatic system may carry cancer cells to other parts of the body. During metastasis, cancer cells grow and may develop into new tumors. The lymph nodes, which are small, bean-shaped organs that play a role in fighting infection, are one of the first places cancer spreads to. Lymph node clusters are situated in different parts of the body (e.g. under the arms). Cancer is also able to spread through the bloodstream to parts of the body located far away. Even within the same tumor, cancer cells can be heterogeneous. Biopsies are naturally taken from only one spot within a tumor, this causes implications when trying to improve diagnostics and therapies. This also means that one targeted therapy is not likely to abolish all cancer cells alone. Cancer cells grow and divide extremely fast and must tolerate a lot of strain and harm to their DNA. Cancers rely on a balance between damage and repair of their DNA. Changes in their genes build up over a period of time causing evolution at a rapid pace leading to mutations. This contributes to the heterogeneity of cancer cells and cancer treated today can differ from cancer you treat in the time to come. A kombucha SCOBY is a symbiotic colony of bacteria and yeast. Using this SCOBY kombucha tea is made consisting of two parts: a sour liquid broth and a pellicle layer of floating cellulose. Tea polyphenols (e.g. catechins), sugars, glucuronic acid, gluconic acid, lactic acid, amino acids (e.g. lysine), fibers, vitamins (e.g. vitamin C, vitamin B), essential elements (e.g. Na, K, Ca, Cu, Fe, Mn, Ni, Zn), catalase, carbon dioxide, antibiotics, and different micronutrients are produced when kombucha tea is fermented and most likely contributes to the health benefits observed when consuming kombucha tea. As fermentation takes place, the yeast component breaks down to sucrose to produce fructose, glucose, and carbon dioxide. Carcinogenesis is a multistage, multifactorial process and several genes are affected. Chemo-preventive agents can target these genes and regulate their intracellular, extracellular, or cell-surface functions. The anticancer properties of black tea polyphenols have been studied in the past. One of the components of kombucha tea is black tea, so does this mean it has anti-cancer properties? There are many references to and claims that kombucha tea has anticancer properties, but it has not been scientifically proven yet. There are reports that carcinoma has been successfully cured by the consumption of kombucha tea and that long-term consumption of kombucha tea increases the immune system's defense against cancer. In this study, kombucha tea was tested for anticancer properties. Kombucha may provide the health benefits of black tea and have health benefits of its own. One of the healthiest beverages is black tea because it consists of a variety of bioactive compounds, including polyphenols, that are powerful antioxidants. Kombucha tea containing black tea consists of the same plant components and presumably has the same health benefits. In previous studies, black tea consumers have a lowered risk of breast, prostate, and colon cancers. Kombucha tea was analyzed in test-tube studies and helped in the prevention of the growth and the spreading of cancer cells because of the high concentration of antioxidants and tea polyphenols. The theory is that tea polyphenols block or suppress the mutation of genes and the growth of cancer cells, promoting the death of cancer cells. Glucose can be converted into gluconic acid and fructose into acetic acid by acetic acid bacteria. Ethanol is produced by the yeast component of kombucha using fructose as a preferred substrate and the bacterial component oxidizes the ethanol into acetaldehyde. The production of ethanol is stimulated by the acetic acid produced and the production of acetic acid is in turn stimulated by the ethanol produced. Some cases of toxicity associated with ingesting large quantities of indecorously prepared kombucha have been reported. These risks can occur when kombucha is over-fermented or home-brewed. Preparing kombucha involves bacteria growing in a liquid that is then consumed. These bacteria are considered to be probiotics, but improperly prepared, harmful bacteria or mold can grow. Improper preparation of kombucha can cause liver problems, the build-up of lactic acid, allergic reactions, and nausea. Proper preparation of kombucha is essential.
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Defining the Research
Research question
Does kombucha tea have the ability to prevent or even treat cancer?
Hypothesis
- Ha: There is an association between cancer and kombucha tea.
- Ho: There is no association between cancer and kombucha tea.
Aim and Objectives
The purpose of this study is to determine the effect kombucha tea has on cancer and if it is able to prevent or even treat cancer.
Methods
Plate Count Method to Isolate a Safe, Pure Culture
2 Vials of the kombucha scoby and 1 vial of the reference stock were placed in a freezer and taken out to warm to room temperature. The cap was disinfected with 70% ethanol. The vials were rehydrated and the contents were allowed to reconstitute for 5 minutes. Using inversion, the vials were mixed thoroughly. A tenfold dilution series of the kombucha scoby was prepared in capped tubes and 9.0ml of the diluent was pipetted into each tube using a 10.0ml pipette and labeled. 1.0ml of the first sample was transferred into the first diluent tube using a pipette, capped, and then mixed. The dilution was continued using a pipette to transfer 1.0ml of the first tube into the second tube. The dilution was repeated 10 times. 0.1ml of tubes 8, 9, and 10 were deposited on 3 different plates of agar with a pipette. An inoculum spreader was sterilized in 70% alcohol. The inoculum spreader was used to distribute the inoculum on the agar surface, sterilizing between each distribution. The plates were inverted and incubated.
Preparation of Fermented Kombucha Tea
3.0 g of green tea, 3.0g of black tea, and 3.0g of tea powder were respectively boiled in 400 ml of water for 20min. Each tea infusion was filtered using filter paper and sucrose was dissolved in the clear filtrates and left to cool. The 200ml of the tea infusions were poured into a sterilized flask. The flask was inoculated with 3% of the pure kombucha culture. The flask was covered with a cloth. Fermentation was carried out in the dark at 30°C for two weeks. At 10 000rpm the fermented tea was centrifuged for 10min.
Preparation of Fermented Control Tea
3.0 g of green tea, 3.0g of black tea, and 3.0g of tea powder were respectively boiled in 400 ml of water for 20min. Each tea infusion was filtered using filter paper and sucrose was dissolved in the clear filtrates and left to cool. The 200ml of the tea infusions were poured into a sterilized flask. The flask was covered with a cloth. Fermentation was carried out in the dark at 30°C for two weeks. At 10 000rpm the fermented tea was centrifuged for 10min
Gelatine Zymography
Preparation of Conditioned Media
The cell cultured in fetal bovine serum was put in a flask. At 80% confluency, the fetal bovine serum media was removed. The cells were washed with fetal bovine serum-free media twice and left to grow. The conditioned media was collected and centrifuged to remove the dead cells and then concentrated.
Running the gel
To ensure that all the samples have the same concentration the conditioned media was diluted. A non-reducing sample buffer was added to the samples. A 7.5% acrylamide gel containing gelatine was prepared. Each sample was loaded into a well and ran at 150V until a good band separation was observed.
Gel washing and staining
The gel was washed two times for 30 minutes with a washing buffer to remove SDS from the gel. The gel was rinsed for 10 min with an incubation buffer. The gel was incubated in a fresh incubation buffer for 24 hours. The gel was stained for 1h and then rinsed with water to remove the excess staining solution. The gel was incubated until the bands were clearly visible.
Study Design
The fermented kombucha tea was given to 10 healthy subjects and 10 cancer-infected subjects. The fermented control tea was also given to 10 healthy subjects and 10 cancer-infected subjects.
Pathology Tests
Pathology tests were performed on all 40 subjects, using a microscope to evaluate the presence of abnormal cells.
Diagnostic Imaging
The abnormal masses of all 40 subjects were visualized using high-tech machines (e.g. x-rays and computed tomography scans) to create images.
Blood Tests
Blood tests were performed on all 40 subjects measuring substances indicating how advanced the cancer is.
Tumor Markers
All 40 subjects were tested to detect substances in the blood, urine, or other tissues that would be higher than normal when cancer was present.
Conclusion
The Plate Count Method was used to isolate a kombucha culture. Using this method increases the chances of obtaining a pure, safe culture, reducing the chances of bacteria that can cause harm to the human body. A control fermented tea was used to establish that the results obtained were true because of the fermented kombucha tea and not because of other factors. Molecular processes such as tumor cell apoptosis and tumor growth and invasion inhibition are affected by tea polyphenol. Matrix MMPs are linked to tumor cell evasion and kombucha tea inhibits MMPs. Gelatine zymography assay showed that kombucha tea reduced MMP activity according to the concentration used. Tea polyphenols demonstrated growth inhibitory activities. Pathology tests, diagnostic imaging, blood tests, and tumor markers were used to observe the presence of abnormal cells or cancer cells before and after the consumption of either the fermented kombucha tea or the control tea. The healthy subjects who consumed the kombucha tea showed an increase in immunity and anti-cancer properties. The healthy subjects who consumed the control tea showed little difference in immunity. The cancer-infected subjects who consumed the fermented kombucha tea showed an inhibition of tumor growth and prevented the cancer from spreading. The cancer-infected subjects who consumed the control tea showed no change in tumor growth.
References
- M. I. Watawana, N. Jayawardena, C. B. Gunawardhana and V. Y. Waisundara, “Health, Wellness, and Safety Aspects of the Consumption of Kombucha,” Journal of Chemistry, 2015.
- C. Fu, F. Yan, Z. Cao, F. Xie, and J. Lin, “Antioxidant activities of kombucha prepared from three different substrates and changes in the content of probiotics during storage,” Food Science and Technology, 2014.
- I. Vina, P. Semjonovs, R. Linde and I. Denina, “Current Evidence on Physiological Activity and Expected Health Effects,” Journal of Medicinal Food, 2014.
- J. M. Leal, L. V. Suárez, R. Jayabalan, J. H. Oros and A. Escalante-Aburto, “A review on health benefits of kombucha nutritional compounds and metabolites,” CyTA - Journal of Food, 2018.
- “What is Cancer?,” February 2018. [Online]. Available: https://www.cancer.net/navigating-cancer-care/cancer-basics/what-cancer.
- W. C. Research, “Why Haven't We Cured Cancer?” 2018. [Online]. Available: https://www.worldwidecancerresearch.org/blog-post/havent-cured-cancer/.
- K. Zhang, “Why is Cancer so difficult to cure,” 2019. [Online]. Available: https://www.jax.org/news-and-insights/2015/december/why-no-cure-for-cancer#.
- “Obtaining A Mother Kombucha SCOBY,” [Online]. Available: https://www.culturesforhealth.com/learn/kombucha/obtaining-a-kombucha-scoby/.
- R. Jayabalan, P.-N. Chen, Y.-S. Hsieh, K. Prabhakaran, P. Pitchai, S. Marimuthu, P. Thangaraj, K. Swaminathan and S. E. Yun, “Effect of solvent fractions of kombucha tea on viability and invasiveness of cancer cells—Characterization of dimethyl 2-(2-hydroxy-2-methoxypropylidine) malonate and vitexin,” Indian Journal of Biotechnology, pp. 75-82, 2011.
- B. Krietsch, “Is Kombucha Healthy? Here’s What Experts Say,” 11 February 2019. [Online]. Available: http://time.com/5516472/is-kombucha-healthy/.
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