Effectiveness of Artemisia Vulgaris as Supplementation Against Chemotherapy of Mammae Adenocarcinoma to Reduce Expression of NF -κB and CD 34 (Study in C3H Mice Given Chemotherapy Adriamycin-Cyclophosphamide)

Intoduction: Breast cancer is a significant healthcare problem worldwide. Surgery remains the treatment of choice combined with other modalities such as chemotherapy, radiation, and immunotherapy such as Artemisia vulgaris (AV). Selective cytotoxicity of AV is intended as a supplementation to AdriamycinCyclophosphamide, improving the response rate of chemotherapy in adenocarcinoma mammae. Method: This study used a "Post-test only control group design" on 24 females C3H mice that were randomly selected and divided into four groups: group K (control), P1 (chemotherapy), P2 (extract), and P3 (combination). Adenocarcinoma mammae come from the inoculation of donor mice. Chemotherapy of Adriamycin 60 mg / m2 and Cyclophosphamide 600 mg / m2 were given in two cycles. AV 13 mg (0.2 ml) was given once daily orally. NF-κB expression and CD34 were evaluated using imunohistochemical staining. Result: The expression of NFκB and microvascular density of CD 34 were obtained in groups of K, P1, P2, P3 Statistical analysis showed significant decrease in the expression of NF-κB between groups K and P1, P2, P3. Correlation analysis between NF-κB expression with CD 34 was found to have significant correlation (p = 0,039 and r = 0,897). Conclusion: Artemisia vulgaris can reduce angiogenesis by decreasing NF-κB expression and the microvascular density CD34 of adenocarcinoma mammae of C3H mice treated with Adriamycin-Cyclophosphamide chemotherapy and can improve the effectivity.

control. In this case, Nuclear Factor-Kappa B (NF-κB) has a vital role in regulating regulation, including processes from inflammatory reactions, growth, and vascularity formation to oncogenesis. (7) Incorrect regulation of NF-κB is associated with the incidence of malignancy. (8) In previous studies, NF-κB was a regulator of cell proliferation and protected cells against conditions that lead to apoptosis.(9) NF-κB binds to deoxyribonucleic acid (DNA) and causes transcription of genes that will cause oncogenesis processes, such as inflammation, anti-apoptosis, and increased cell proliferation activity metastasis, and angiogenesis. (8,10) Angiogenesis factor plays a vital role in the growth of cancer cells, progression, and metastasis. The   (19).
The mechanism of action of artemisinin compounds as anticancer is anti-angiogenic, anti-inflammatory, anti-metastatic, and inhibiting cancer cell growth. It is known that artemisinin contains an endoperoxide moiety that can react with iron to form cytotoxic free radicals (20,21).
Although it is toxic, artemisinin has an advantage of being used as an anticancer because it has particular toxic properties. (20) This is an essential consideration in terms of safety for its users.

Research design
This research is an experimental laboratory study with the design "Post-test only control group design." The research group was divided into 4, namely the control group (K), treatment 1 (P1), treatment 2 (P2), and treatment 3 (P3

Research variable
The independent variable in this study was the administration of Artemisia vulgaris extract with a combination of chemotherapy Adriamycin and Cyclophosphamide.
The dependent variables in this study were the expression of NF-κB and the microvascular density CD34.  Tumors from donor mice will be incised with a biopsy, and a histopathological examination will be performed to confirm the type of tumor.

Data analysis
After the data was collected, data coding, and tabulation was carried out. The data was processed and presented in tables and box plots to see the distribution of data.

Ethical requirements
The research applies animal ethics and was approved by the Ethics Commission for Health Research, Faculty of Medicine, Diponegoro University.

Descriptive analysis
Description of NF-κB expression data The highest mean NF-κB expression was found in the K group, while the lowest average NF-κB expression was found in the P3 group.

Discussion
The first hypothesis test obtained an F count of Pharmacologically, Cyclophosphamide is an alkylating agent. This agent will be metabolized in the liver and produce phosphoramide mustard metabolites which will bind to DNA and cause DNA to become damaged, which will cause the cell to fail in mitosis and cell death. are significant regulators of the angiogenesis process.
The blockade of the NF-κB pathway by artemisinin against the VEGFR2 promoter will inactivate transcription activity so that proteins that play a role in angiogenesis are not formed. (39) The expression of NF-κB and CD34 has a very strong relationship. NF-κB is a protein that functions to regulate the formation of blood vessels when they translocate into the cell nucleus. The transcription factor NF-κB is activated when there are proinflammatory cytokines including TNF-α, which will activate NFκB, causing VEGF production and VEGFR expression to increase with clinical parameters in the form of increased CD34 microvascular density (25,40,41).
The results showed that the administration of Artemisia vulgaris extract had a synergistic effect on the administration of AC chemotherapy in a population of C3H adenocarcinoma mammae mice. This suggests that Artemisia vulgaris can be given as a supplement to AC chemotherapy.

Conclusion
Artemisia vulgaris extract improves chemotherapy response in breast adenocarcinoma of C3H mice given

Conflict of interest and funding.
The author has not received funding or profits from the industry or elsewhere for conducting this research.