ANTICANCER EFFECT OF Citrus hystrix LEAF EXTRACTS ON THE TRIPLE NEGATIVE BREAST CANCER CELL LINE MDA-MB-231

Abstract

Triple-negative breast cancer is one of the most aggressive types of breast cancer due to the ability of early metastasis and chemoresistance, which accounts for almost 10-15% of invasive breast cancer. Citrus hystrix DC. is a tropical plant known for its high aroma. It has been used as spices, in cosmetic products and also has been reported to possess many biological activities including anticancer effects. However, the effect of C. hystrix against triple-negative breast cancer has yet been identified. In this study, cytotoxicity of C. hystrix leave extracts, and its bioactive compounds citronellal and citronellol against the triple-negative breast cancer MDA-MB-231 cell line were observed. C. hystrix leaves powder was sequentially extracted using maceration with hexane, ethyl acetate and ethanol. Cytotoxic effect of extracts were evaluated using MTT assay. Cell proliferation, wound scratch migration, colony formation, and apoptosis assay were used to identify the in vitro effect of the extracts and its active compounds against this cell line. RT-qPCR and Western blot analysis were used to understand some mechanism which may lead to the cause of cells dead. The results significantly showed that crude hexane of C. hystrix leaves extract, and its bioactive molecules could reduce cell proliferation, colony forming, migration of the cancer cells, and induced cell cycle arrest when compared to control. The results also indicated that crude hexane, citronellol and citronellol could induce apoptosis in MDA-MB-231 cells through the inhibition of the anti-apoptotic Bcl-2 expression and leading to the activation of the caspase-dependent pathway through the activation of the executioner Caspase-3. This is the first demonstration of the effect of C. hystix and its bioactive compounds citronellol and citronellal against the triple negative breast cancer MDA-MB-231 cells. Further study should be held to help understanding more deeply on mechanism that caused by the extract and the molecules on cells.

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