**1. Introduction**

Cervical cancer is known to be the fourth most common malignancy, leading to a substantial burden and threatening women's health worldwide [1]. A persistent infection by human papilloma virus (HPV), especially HPV16 and 18, is critical for cervical cancer initiation and progression [2,3]. The implementation of HPV vaccines, HPV testing, and cervical cytology has notably decreased the incidence of early-stage cervical cancer [2,4,5]. However, patients with advanced cervical cancer still have unfavorable outcomes due to the high incidence of metastasis, which is still one of the main factors influencing the prognosis of patients. Moreover, the clinical application of immunotherapy or adjuvant chemotherapy approaches in advanced cervical cancer has not been as effective as once expected [6,7]. Therefore, there is an urgent need for novel therapeutic approaches.

Cervical cancer is a complicated disease involving the disruption of normal complex biological networks in the human body [8]. Upon HPV infection, several key molecular events are involved in the initiation and progression of cervical cancer, including the

**Citation:** Chen, T.; Wang, J.; Li, M.; Wu, Q.; Cui, S. Genistein Inhibits Proliferation and Metastasis in Human Cervical Cancer Cells through the Focal Adhesion Kinase Signaling Pathway: A Network Pharmacology-Based In Vitro Study in HeLa Cells. *Molecules* **2023**, *28*, 1919. https://doi.org/10.3390/ molecules28041919

Academic Editors: Barbara De Filippis, Alessandra Ammazzalorso and Marialuigia Fantacuzzi

Received: 4 January 2023 Revised: 7 February 2023 Accepted: 7 February 2023 Published: 17 February 2023

**Copyright:** © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).

Toll-like receptor (TLR) pathway, the nuclear factor-kappa B (NF-κB) pathway, the Notch signal pathway, the Hippo-Yes-associated protein (YAP1) pathway [9–11], and the focal adhesion pathway [12]. However, drugs that act on single-molecular targets usually exert an unsuccessful effect or show strong toxic side effects in clinical practice. Considering the complexity of the pathogenesis of cancers, attention has switched from focusing on single drug targets towards a more systemic view of drug targets [13]. Network pharmacology is considered a powerful tool in deciphering the complexity of biological systems and provides a new concept for understanding the interplay of molecular networks of compounds [14]. This approach is now widely used in the study of the pharmacological effects of compounds, where it promotes new directions for efficient drug discovery.

Genistein is a well-studied natural flavone compound with a wide range of biological effects, including tyrosine kinase inhibitory, anti-inflammatory, phytoestrogen, and anticancer effects [15–20]. Several studies have already been published concerning genistein treatment during cervical cancer development, and the reported activities of genistein include inducing cell cycle G2-M phase arrest and apoptosis, inhibiting the action of histone deacetylases and DNA methyltransferases, and synergizing the radiation effect by cell cycle G2-M phase arrest and AKT (Protein Kinase B) activation [21–26]. Considering the range of activities identified in the above-mentioned studies, the precise molecular action of genistein against cervical cancer remains largely unknown. Therefore, in order to investigate the molecular action of genistein against cervical cancer more comprehensively, this study employed an integrated strategy that combined the network pharmacology approach and RNA-seq analysis to identify critical targets related to the genistein treatment of cervical cancer.
