The Cytotoxic and Anti-Migratory Properties of Caesalpinia sappan and Ficus septica, in Combination with Doxorubicin on 4T1 TNBC Cells with Nephroprotective Potential

Abstract

Objective: To evaluate the anti-cancer properties of Caesalpinia sappan and Ficus septica in combination with doxorubicin on 4T1 cells, confirm their nephroprotective activities, and predict the molecular targets of the underlying mechanisms. Methods: The cytotoxic activities of all extracts and doxorubicin were determined by MTT assay followed by cell cycle and apoptosis analysis using flow cytometry. Immunoblotting was used to determine the protein expressions. The proteins involved in the cell proliferation and migration were analyzed through bioinformatics approaches, whereas, the interaction between compounds and protein targets was observed through molecular docking. Furthermore, the effect of the extracts on cell migration was analyzed by scratch wound healing assay. The intracellular ROS after treatment with extracts was observed using DCFDA staining flow cytometry. Results: Both ECS and EFS performed cytotoxic properties and significantly enhanced doxorubicin's cytotoxic effects against 4T1 cells. However, these cytotoxic activities did not correlate with the cell cycle progression. On the contrary, the combination treatment caused apoptosis that may correlate with the decreasing of IκBα phosphorylation, indicating that all agents targeted the inhibition of NF-κB activation. The combination treatments also inhibited cell migration and decreased MMP-9 expression. TNBC proliferation and metastasis needed at least 54 proteins to be activated, some of them are related to NF-κB activation. The inhibitory effect of ECS correlated with the interaction of brazilin and brazilein to IKK, a kinase protein that plays a role in IκBα phosphorylation. In addition, ECS and EFS reduced ROS expression in Vero cells caused by doxorubicin. Conclusion: In conclusion, ECS and EFS effectively enhanced the cytotoxic effect of doxorubicin and inhibit cell migration on 4T1 cells and these activities may correlate to the inhibitory effect of NF-κB activation. ECS and EFS also exhibit ROS suppressing effect on Vero cells that may be beneficent to reduce nephrotoxicity of chemotherapeutic treatment.

Cardamom Essential Oil Extract Suppress the Progression of Triple-Negative Breast Cancer 4T1 Cell Line

“…When combined with chemotherapeutic drugs such as Paclitaxel and Vemurafenib, Navitoclax, a Bcl-2 inhibitor, increases cell Metastasis involves some mechanisms in cancer cells, starting from extracellular matrix disruption by the family of MMPs, cell adhesion, and invasion of a cancer cell on the other tissues, which mainly correlated with cell migration. (19,20) This process continued until the cell reached the second host tissue and completed the metastasis process. (20,31,32) However, extracellular matrix disruption in cancer was regulated by MMP-9.…”

“…(1,3) Using these characteristics as the basis, we explored the potential influence of the CEO on TNBC senescence and cell migration. Our research group reported that sappan wood extract (CSE) decreases cell senescence and inhibits TNBC cancer cell line migration (17,19), manifested by the main contents in sappan wood ethanolic extract, brazilin, and brazilein (20,21). Meanwhile, the activity of CEO on the TNBC cell line, including its senescence induction or metastasis inhibition activity, has yet to be studied.…”

Article review: Brazilin as potential anticancer agent

“…An in silico study predicted several targets that brazilin inhibited. In triple breast cancer, brazilin was found to attach more firmly and effectively to MMP-2, MMP-9, and PTGS2 compared to its native ligand (Haryanti et al, 2022). These enzymes are involved in cell migration and metastasis (Webb et al, 2017;Ercolano et al, 2019).…”

Antioxidant, Cytotoxic Activity and Protein Target Inhibition of Ethyl Acetate Fraction Melinjo Seed (Gnetum gnemon L.) by In Vitro and In Silico Studies on HeLa Cervical Cancer Cells

“…The cytotoxic evaluation was conducted based on the metabolically active cells to convert 3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide (MTT) into an oxidative form of formazan that can be detected using visible light (Haryanti et al 2022). The cytotoxic test of the Ethyl Acetate Fraction of Melinjo Seeds (EAFMS) on HeLa cancer cells was carried out by inserting cells with a density of 300 cells/6,700 µl MK into 96 well plates of 100 µl each and providing 3 empty wells to be used as controls, then incubated for 48 hours at 37°C.…”

“…According to Haryanti et al (2022), after incubation, discharging the medium, and washing with PBS (Sigma), each well received 0.5 mg/ml MTT (Sigma) in the medium and was incubated for 3-4 h, followed by adding 10% SDS in 0.01 N HCl and incubated overnight without light. Then viewed under an inverted microscope, purple formazan crystals formed, indicating that the surviving cells react with MTT.…”