ABSTRACT: Triple-negative breast cancer (TNBC) is one breast cancer with high mortality due to aggressive tumor behavior and limited treatment options. Natural products are studied for their potential to be an alternative to combining with cancer drugs to improve treatment efficacy. Plant phytoconstituents can regulate cellular processes such as proliferation, differentiation, invasion, angiogenesis, or migration. Kalanchoe pinnata has been used in traditional medicine, it contains flavonoids, phenols, alkaloids, glycosides, bufadienolides, and tannins among others. In the present investigation, we evaluated the effect of aqueous extract and underlying molecular mechanisms on two breast cancer cell lines, HCC1937 and MDA-MB-231, focusing on cellular migration. The cytotoxic activity revealed that HCC1937 cells exhibit more sensitivity than MDA-MB-231 cells. Cell migration was affected in HCC1937 cells after 24h of treatment compared to 48h in MDA-MB-231 cells. The extract had more anti-invasion properties on HCC1937 cells (37.7%, p<0.01) than MDA-MB-231 cells (47%, p<0.05). RNAseq assay showed 1850 differentially expressed genes (DEGs) in HCC1937 cells versus 1534 in MDA-MB-231 cells. The Webgestalt analysis revealed the downregulation of genes involved in cell-cell adhesion, ruffle organization, cell-substrate junction with more downregulated genes in HCC1937 cells. The downregulated genes analyzed with KOBAS-i showed association with MAPK signaling, Focal adhesion, PI3K-Akt signaling, Regulation of actin cytoskeleton, and ECM-receptor interaction pathway. In addition, IPA network analysis revealed the inhibition of signaling pathways RHO GTPase cycle, RHOA signaling, JAK/STAT signaling, Actin cytoskeleton signaling, and Integrin signaling. Data for the docking study indicated the binding potential of Quercetin-3-O-arabinoside, Kaempferol, Quercetin, Ferulic acid, alpha-L-rhamnopyranoside, and Apigenin with JAK2, ROCK1, ROCK2, and PIK3CA proteins. Thus, our results demonstrate that K. pinnata inhibits the proliferation and invasion of TNBC cells by targeting genes involved in these important processes, so it has the potential to be used in combination with chemotherapeutic drugs for cancer treatment.
