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Published on: May 20, 2015
In Vitro Anti-Breast Cancer Effects of Tamarix aphylla-Derived Quercetin and In Silico Insights into Its Targeting of
Dhurgham Al-Fahad1, Zahraa Naeem Hashim1, Suliman A Almahmoud2
1Department of Pathological Analysis, College of Science, University of Thi-Qar, Nasiriyah 64001, Iraq.
Abstract:
Phosphatidylinositol 5-phosphate 4-kinase type 2 alpha (PIP4K2A) is a key oncogenic driver that regulates the PI5P/PIP2 axis to promote metastatic migration in breast cancer. This study aimed to investigate the therapeutic potential of a crude extract from Tamarix aphylla against breast cancer progression and identify its primary active constituents. The crude extract was initially evaluated against MDA-MB-231 and MCF7 breast cancer cell lines using wound healing assays. Bioassay-guided isolation and screening were deployed to isolate individual components, and the most potent lead compound was structurally characterized using preparative HPLC and FTIR. To analyze its interaction with PIP4K2A, in silico molecular docking, MM/GBSA calculations, and 200 ns molecular dynamics simulations were conducted. In vitro validation was subsequently performed via dose-dependent cytotoxicity assays, scratch assays, single-cell tracking, and RT-qPCR expression analysis. Quercetin was identified as the most potent lead inhibitor against PIP4K2A. Computational modeling revealed that quercetin binds tightly within the PIP4K2A ATP-binding pocket, yielding a superior binding affinity of -10.77 kcal/mol and enhanced thermodynamic stability (ΔGMM/GBSA = -42.6 ± 2.1 kcal/mol) compared to the native ligand (ΔG MM/GBSA = -23.3 ± 1.8 kcal/mol). Molecular dynamics simulations confirmed an induced-fit structural transition that locked the complex into an ultra-stable conformation within a deep global energy minimum basin (-10.8 kcal/mol). In vitro assays demonstrated dose-dependent cytotoxicity, with aggressive triple-negative MDA-MB-231 cells exhibiting higher sensitivity (IC50 = 82.23 µg/mL) than luminal MCF7 cells (IC50 = 97.14 µg/mL). Furthermore, scratch and single-cell tracking assays showed a profound suppression of migration speed and wound closure (reduced to ~40%), while RT-qPCR revealed a near-complete transcriptional knockdown of PIP4K2A mRNA expression (down to 0.025-fold). Collectively, these findings elucidate a unique dual-action mechanism for Tamarix aphylla-derived quercetin-characterized by both direct competitive enzymatic inhibition and downstream transcriptional silencing-positioning it as a promising therapeutic scaffold for targeted anti-metastatic breast cancer interventions.
Insights
Quercetin from Tamarix aphylla inhibits breast cancer metastasis by targeting PIP4K2A. This natural compound offers a dual mechanism of action, blocking enzyme activity and reducing gene expression for potential anti-metastatic therapies.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Phosphatidylinositol 5-phosphate 4-kinase type 2 alpha (PIP4K2A) drives breast cancer metastasis by regulating the PI5P/PIP2 axis.
- Targeting PIP4K2A presents a therapeutic strategy for inhibiting cancer cell migration.
Purpose of the Study:
- To investigate the anti-cancer potential of a Tamarix aphylla crude extract against breast cancer.
- To identify active constituents targeting PIP4K2A and elucidate their mechanism of action.
Main Methods:
- Bioassay-guided isolation of active compounds from Tamarix aphylla extract.
- In silico molecular docking, MM/GBSA, and molecular dynamics simulations to assess quercetin-PIP4K2A interactions.
- In vitro validation using cytotoxicity, scratch, single-cell tracking, and RT-qPCR assays.
Main Results:
- Quercetin identified as a potent PIP4K2A inhibitor with strong binding affinity (-10.77 kcal/mol) and stability.
- Molecular dynamics confirmed an ultra-stable quercetin-PIP4K2A complex.
- In vitro studies showed quercetin's dose-dependent cytotoxicity, suppressed migration (~40% reduction), and significant PIP4K2A mRNA knockdown (0.025-fold).
Conclusions:
- Tamarix aphylla-derived quercetin exhibits a dual mechanism against breast cancer metastasis: direct enzymatic inhibition and transcriptional silencing of PIP4K2A.
- Quercetin represents a promising therapeutic scaffold for targeted anti-metastatic breast cancer interventions.
