Effects of Alantolactone on Stemness Genes Expression in the Epithelial Mesenchymal Transition (EMT) in Breast Cancer

Background: To reduce the side effects of conventional chemotherapy drugs, new herbal remedies, such as alantolactone, can be used. Objective: We evaluated the association between stemness and EMT (epithelial-mesenchymal transition) process in triple-negative breast cancer cells treated with alantolactone that targets STAT3. Methods: The MDA-MB-231 cell line was used as one of the triple-negative breast cancer cell lines. MTT assay was used to evaluate cell viability and drug dose at three-time points of 24, 48, and 72 hours, and three doses, i.e., 1, 0.1, and 0.01 mM of alantolactone were used to evaluate cellular behavior in proliferative and invasion pathways, respectively. A scratch test was also performed to evaluate the invasive power of cancer cells. Real-time PCR was used to evaluate the expression of regular genes by cancerous cell proliferation, STAT3 NANOG, SOX-2, and E-cadherin metastasis. Results: It was observed that increasing the dose of alantolactone increased cell apoptosis rate. The three doses selected for the cell culture study did not differ significantly from the control group regarding apoptosis-inducing abilities at desired time intervals. In the scratch test, the least amount of repair was seen at 1 mm. Expression of SOX-2, STAT3, and NANOG in the treated cells decreased with increasing dose of the drug, whereas expression of E-cadherin was found to be increased. Conclusion: Alantolactone through the STAT3 signaling pathway affects the expression of Ecadherin, NANOG, and SOX2 genes, inhibiting the EMT process and subsequent stemness, and may potentially be used in therapeutics for cancer patients.