Anti-Cancer and Anti-Proliferation Activity of Ethyl Asetat Extract From Ant Nest (Myrmecodia pendans) in Burkitt’s Lymphoma Cancer Cells

Objective: To determine the activity of anti-cancer and anti-proliferation of ethyl acetate fraction of ant nest plants (Myrmecodia pendans) in Burkitt’s Lymphoma cancer cells. Material and Methods: The study was conducted in a pure laboratory experimental method using Burkitt's Lymphoma cancer cell culture. Gradual research begins with the determination, extraction and fractionation of ant nest plants, to test for proliferation barriers. Data analysis using two-way ANOVA followed by Post Hoc LSD test with a significance level of 95%. Pearson correlation test was conducted. Results: The results of testing the inhibition of Burkitt’s Lymphoma cell proliferation with ethyl acetate extract treatment showed that there was inhibition of cell growth based on the concentration given, starting from the lowest concentration of 15.625 μg/mL. Likewise, the incubation time factor of 24, 48, and 72 hours showed that the longer the incubation time, the greater the inhibition of cell growth. Antiproliferation analysis of flavonoid ethyl acetate extract based on concentration and incubation time on absorption of optical density Burkitt’s Lymphoma was statistically significant (p = 0.00). Conclusion: Ant nest ethyl acetate extract has the effect of proliferation inhibition on Burkitt’s lymphoma cells.


Introduction
Cancer is a deadly disease and cause of death in industrialized countries and the second cause of death in developing countries. Cancer is a non-communicable disease, which is characterized by abnormal/persistent, and uncontrolled cell growth, which can damage the surrounding tissues and can spread to places far from their origin called metastasis. According to WHO data in 2013, the incidence of cancer increased from 12.7 million cases in 2008 to 14.1 million cases in 2012, with the number of deaths increasing from 7.6 million people in 2008 to 8.2 million in 2012 [1,2].
Cancer that can cause enlarged lymph nodes is called lymphoma. Lymphoma is a general term for various types of blood cancers that appear in the lymphatic system. According to the 2012 GLOBOCAN (IARC) data, lymphoma is one of the ten most cancers in the world in 2012.
Lymphoma is divided into two types, namely Hodgkin's Lymphoma and Non-Hodgkin's Lymphoma.
About 90% of lymphoma patients are patients with Non-Hodgkin Lymphoma, and the rest are Hodgkin's Lymphoma [3].
Non-Hodgkin's lymphoma occurs due to mutations that occur in the immune system caused by infectious agents, carcinogenic substances and a history of other diseases suffered by a person [4].
Burkitt's Lymphoma is a type of Non-Hodgkin Lymphoma. Burkitt's Lymphoma is a mature neoplasm of B lymphocyte cells and includes aggressive lymphoma. Burkitt's Lymphoma is a malignancy of B-lymphocytes that can be cured and is first known to be associated with HIV.
Burkitt's Lymphoma is also one of the lymphoid tumors characterized by chromosomal translocation, especially translocation on the MYC, which is characteristic of Burkitt Lymphoma.
Burkitt's Lymphoma plays an important role in the mechanism of carcinogenesis and lymphomagenesis. Several studies have found that Burkitt's Lymphoma is most likely derived from B-lymphocytes in the germinal flashlight (GC) [5].
Clinically, Burkitt's Lymphoma generally occurs in children. The incidence occurs at the age of 3 to 8 years, where men are more susceptible to 2x than women. Lesions are commonly found in the maxilla, mandible and abdomen. The most typical sign in the oral cavity of this disease is the presence of local tumors and tooth mobility. Symptoms are localized, blunt and paresthesia pain.
Clinical symptoms of Burkitt's Lymphoma are initially only recognized by enlarged lymph nodes without pain and grow rapidly on the neck, thighs, under the jaw or it can also be under the hand. In the sporadic type, a lump begins in the central or abdominal area [6].
The results of the study have shown that ant nest plants are one of the medicinal plants that are believed to have potential effects in the world of health. Although modern therapies such as chemotherapy give positive results in the treatment of cancer, on the other hand many cause side effects. Therefore, herbal treatment is often a cancer treatment option. In addition to the low cost, the side effects produced are also minimal compared to modern therapies [7,8].
Chemical screening tests for ant nest plants indicate that these plants contain flavonoid and tannin class chemicals. Based on the results of this research, this plant also contains active compounds of tocopherols, phenols, and is rich in various useful minerals [9,10]. Many working mechanisms of flavonoids have been revealed, such as inactivation of carcinogens, antiproliferation, cell cycle inhibition, apoptosis induction and differentiation, and inhibition of angiogenesis [11].
Flavonoids in the human body function as antioxidants so it is very good for cancer prevention. Flavonoids have a carbon base frame consisting of 15 carbon atoms, where two benzene rings (C6) are bound to a propane chain (C3) so that they form a C6-C3-C6 arrangement. This arrangement can produce three types of structures, namely 1,3-diarylpropane or flavonoid, 1,2diarylpropane or isoflavonoids, and 1,1-diarylpropane or neoflavonoids [12]. In some cases, flavonoids can act directly as antibiotics by disrupting the function of microorganisms such as bacteria or viruses.
In vivo research shows that the influence of flavonoid diets on cancer development is: First, it inhibits protein kinase activity. Before the enzyme activity is excessive, the role of flavonoids is needed to prevent the formation of cancer cells, namely by preventing the joining of carcinogen compounds generated by the kinase enzyme with DNA, so that the DNA does not experience damage (cancer). Second, have anti-proliferation activity. Third, it induces apoptosis. Now, only a few potential anti-cancer agents such as flavonoids are known to cause apoptosis. Fourth, it inhibits metastasis/migration/angiogenesis [13,14].
This study was intended to analyze and identify the effect of flavonoid ethyl extract on ant nests (Myrmecodia pendans) as an anticancer against the inhibition of proliferation in Burkitt's Lymphoma cells.

Results
The results showed a significant relations between how much concentration of flavonoid ethyl acetate extract from ant nest plants, incubation time, and how many cells that can survive after being treated with anthyl acetate extracts of ant nests in certain concentrations and times. There was a significant decrease in the number of Burkitt's Lymphoma cells after being treated. For example, at 24 hours of incubation time, the concentration of ethyl acetate extracts of ant nests was 15,625 µg/mL, while the number of cells that survived was less than 60 cells. The number of cells that survive will decrease with increasing ethyl acetate concentration (Table 1).
Based on the results obtained (Figure 1), it is seen that in general flavonoid ethyl acetate extract has Burkitt's Lymphoma cell growth inhibitory activity. This is shown in cell measurements using an ELISA reader.
Research also shows that quercetin, a flavonoid compound, can increase cyclic AMP levels and can also reduce DNA, RNA, and protein synthesis in tumor cells Ehrlich ascites. Quercetin was also found to inhibit aerobic glycosis in tumor cells. Quercetin can also inhibit the activity of specific protein kinases -tyrosine, which play an important role in malignant fibroblast transformation in sarcoma cells. Enzyme inhibitory activity by flavonoids is an anti-proliferative effect on malignant cells, for example in cases of gastritis in humans and colon cancer cells, found inhibition of cancer cell growth by quercetin [18].

Conclusion
Ant nest ethyl acetate extract has anti-cancer activity and anti-proliferation in Burkitt's Lymphoma Cells. Ethyl acetate extract at a concentration of 15.625 mg/mL to 500 pg/mL showed an inhibitory effect Burkitt's Lymphoma Cell growth. Large concentrations of the extract and the duration of incubation determines the inhibition of Burkitt's Lymphoma Cell growth.
Financial Support: None.