3 Inhibition of Cell Migration and Invasion

Tumor metastasis is the leading cause of cancer-related deaths [24]. Migration and invasion constitute key biological processes of tumor metastasis, with distinct mechanisms. Migration refers to the directed movement ability of tumor cells after detaching from the primary site, driven by chemokines or mechanical signals (such as matrix stiffness gradients); in contrast, invasion focuses on the destructive infiltrative behavior of cells penetrating the basement membrane or extracellular matrix, which requires the secretion of proteolytic enzymes (such as matrix metalloproteinases, urokinase-type plasminogen activator) and the downregulation of adhesion molecules mediated by epithelial-mesenchymal transition (EMT) to complete synergistically. In vitro studies on the migration and invasion abilities of prostate cancer cells are important means to evaluate their invasiveness [25].

 

CISTANCHE HEALTH SUPPLEMENTS 

 

 

 

3.1 Promote Tumor Necrosis Factor Receptor-Associated Factor 6 (TRAF6) Degradation to Reverse Epithelial-Mesenchymal Transition

As a member of the E3 ubiquitin ligase family, TRAF6 plays an important role in inflammation, immune response, and tumor progression by regulating signaling pathways such as nuclear factor-κB (NF-κB) and MAPK. Khusbu et al. [26] treated androgen-independent human prostate cancer DU145 and PC3 cells with 50 μmol/L resveratrol for 48 hours, and found through Transwell migration assay, scratch assay, and Western blot analysis that resveratrol significantly inhibits the proliferation and migration of prostate cancer cells by inducing ubiquitination-dependent degradation of TRAF6. Its specific mechanisms include: depletion of TRAF6 leads to decreased Akt phosphorylation (blocking cell cycle progression), inhibits NF-κB nuclear translocation (reducing levels of pro-inflammatory factors IL-6 and TNF-α), and reverses the EMT process (including upregulation of E-cadherin and downregulation of mesenchymal markers such as N-cadherin and Snail), thereby impairing the invasive and metastatic potential of prostate cancer cells. This study reveals a new anti-cancer mechanism of resveratrol targeting TRAF6, providing support for natural products in the treatment of prostate cancer. Notably, studies have found that reducing the ubiquitination level of TRAF6 can promote the proliferation, migration, and invasion of gastric cancer cells [27]. These studies indicate that the function of TRAF6 in tumors is dependent on tissue type, and resveratrol may overcome TRAF6 functional heterogeneity through multi-target and multi-pathway synergistic effects.

It is worth mentioning that Cistanche Tubulosa, another promising herbal for prostate health, also exerts anti-inflammatory effects by reducing TNF-α and IL-6 levels [24], which are key inflammatory factors involved in tumor migration and invasion. This overlap suggests that both resveratrol and Cistanche Tubulosa target core inflammatory pathways, making them potential combined candidates for herbal prostate therapies.

 

CISTANCHE HEALTH SUPPLEMENT

3.2 Inhibit the Akt/miR-21 Signaling Pathway

The Akt/miR-21 signaling pathway plays a crucial role in tumor occurrence and development. Akt promotes tumor cell survival, proliferation, and drug resistance through interaction with miR-21. Programmed cell death protein 4 (PDCD4) is a tumor suppressor protein that inhibits tumor occurrence and metastasis by regulating gene translation and cell apoptosis. Mammary serine protease inhibitor (Maspin) is a tumor suppressor belonging to the serine protease inhibitor (serpin) family, which inhibits tumor invasion, metastasis, and angiogenesis through non-enzymatic mechanisms, and is lowly expressed in prostate cancer cells [28]. Sheth et al. [29] used the highly invasive androgen-independent human prostate cancer PC-3M-MM2 cell line, supplemented with DU145 and LNCaP cells for comparative analysis. In in vitro experiments, cells were treated with 5-100 μmol/L resveratrol for 24-72 hours; in in vivo experiments, a SCID mouse subcutaneous xenograft model was used, and 20 mg/kg resveratrol was administered intragastrically. The results revealed that resveratrol can reduce the phosphorylation level of Akt in invasive prostate cancer cells (PC-3M-MM2), thereby downregulating the expression of oncogenic miR-21. This downregulation relieves the inhibition of miR-21 on tumor suppressors PDCD4 and Maspin, thereby inhibiting cancer cell proliferation, migration, and inducing cancer cell apoptosis. The results indicate that resveratrol can inhibit the growth and metastasis of prostate cancer by inhibiting the Akt/miR-21 signaling pathway.

3.3 Regulate the Androgen Receptor/CXC Chemokine Receptor 4 (CXCR4) Axis

The occurrence, development, and treatment of prostate cancer are closely related to the androgen receptor (AR) [30]. Reprogramming of AR signaling can promote the EMT process of prostate cancer, thereby enhancing the migration, invasion ability of cancer cells, and the tendency to metastasize to bones. CXCR4 is a G protein-coupled receptor whose ligand is CXCL12 (SDF-1), and plays a role in tumor cell proliferation and metastasis [31]. Jang et al. [32] used a hormone-stimulated LNCaP cell model constructed by treating with 0.1 μmol/L dihydrotestosterone (DHT) for 7 days, and then intervened with 50 and 100 μmol/L resveratrol. The results showed that DHT can drive cell proliferation, migration, and EMT by activating the AR-CXCR4 axis, while resveratrol can upregulate the expression of E-cadherin and downregulate the levels of N-cadherin and vimentin in LNCaP cells, thereby reversing the EMT phenotype. This indicates that resveratrol inhibits DHT-induced migration of prostate cancer cells by regulating the AR/CXCR4 pathway [32].

Similar to resveratrol's regulation of hormonal balance, Cistanche Tubulosa has been shown to enhance testosterone production by upregulating key proteins (StAR, CYP11A1, and HSD17β3) involved in hormone synthesis [24]. By restoring hormonal homeostasis, Cistanche Tubulosa may indirectly modulate AR-related signaling pathways, complementing resveratrol's direct regulation of the AR/CXCR4 axis in prostate cancer intervention.

 

3.4 Bidirectional Regulation of the Tumor Microenvironment

To explore the molecular mechanism of high metastatic potential in prostate cancer, Hsieh et al. [33] used a co-culture model of human prostate stromal cells (PrSC) and epithelial cancer cells (LNCaP, DU145, PC-3). On the one hand, conditioned medium was prepared by treating PrSC cells with 25-50 μmol/L resveratrol for 1-3 days; on the other hand, cancer cells were treated with the same concentration for 2-7 days. The molecular mechanism of resveratrol inhibiting prostate cancer metastasis was studied by combining Boyden chamber migration assay, time-lapse microscopy, and molecular biology techniques. The results found that resveratrol can target and inhibit the secretion of hepatocyte growth factor (HGF) by prostate stromal cells, block the activation of the HGF/c-Met signaling pathway, significantly upregulate the level of E-cadherin, and reverse the EMT process. Experimental data showed that resveratrol can reduce cell migration ability by 60%-80% and decrease matrix penetration activity. Previous studies have found that resveratrol has a regulatory mechanism on the tumor microenvironment, while this study proposes a bidirectional regulatory mechanism of resveratrol on the tumor microenvironment, providing theoretical support for the development of new therapeutic strategies targeting the pre-metastatic microenvironment of prostate cancer [6, 33].

3.5 Inhibit Vasculogenic Mimicry Formation

Vasculogenic mimicry (VM) is a unique phenomenon in which tumor cells mimic vascular structures, and its molecular mechanism involves the activation of EMT. Vascular endothelial cadherin (VE-cadherin) is a member of the calcium-dependent adhesion molecule family, specifically expressed in vascular endothelial cells, and directly involved in the formation and function of VM. VM is closely related to the high invasiveness, metastatic potential, and poor prognosis of tumors. Therefore, targeted strategies for VM-related pathways have become a new direction in tumor drug therapy [34]. Scholars used human prostate cancer PC-3 and DU145 cells as models, treated them with 10-40 μmol/L resveratrol for 24 hours, and used serum as an inducer to explore the inhibitory effect of resveratrol on VM. The experiment found that resveratrol can regulate the EphA2/twist-VE-cadherin/Akt signaling pathway, inhibit serum-induced VM in prostate cancer PC-3 cells, and thereby reduce their metastatic ability. Specifically, resveratrol can significantly inhibit the formation of VM tubular structures in prostate cancer PC-3 cells, and reduce the expression of VE-cadherin at both mRNA and protein levels [35]. This study provides new molecular mechanism evidence for resveratrol as a potential anti-tumor metastasis drug, indicating that it may overcome the drug resistance of traditional anti-angiogenic drugs by targeting the VM process.

These studies indicate that resveratrol synergistically inhibits the migration, invasion, and metastasis of prostate cancer cells through multiple mechanisms. In the future, proximity labeling technology and mass spectrometry can be combined to dynamically capture the ubiquitination modification and interacting protein networks of key targets such as TRAF6 and VE-cadherin, screen potential regulatory nodes, and further analyze the mechanism of resveratrol inhibiting prostate cancer metastasis. For developers focusing on herbal prostate therapies, the multi-faceted regulation of the tumor microenvironment and metastatic pathways by resveratrol, combined with Cistanche Tubulosa's ability to reduce oxidative stress and inflammation [24], offers a promising direction for developing comprehensive herbal formulations.

 

4 Intervention in Epigenetic Reprogramming

Epigenetics involves reversibly affecting gene expression through chemical modifications (such as DNA methylation, histone modification) and regulation of non-coding RNAs without changing the DNA sequence, and is a key factor driving tumor occurrence, metastasis, and drug resistance [36]. As a natural active ingredient, resveratrol can participate in epigenetic modifications by activating sirtuin 1 (SIRT1) [37].

 

4.1 Downregulate the Expression of Metastasis-Associated Protein 1 (MTA1)

As a key node in epigenetic regulation, MTA1 can drive the development and metastasis of prostate cancer by forming MTA1/HDAC complexes, regulating Epi-miRNAs networks, and promoting inflammatory signaling [38]. Kumar et al. [39] used human prostate cancer LNCaP (androgen-sensitive), DU145 (androgen-independent), and PC3M (highly metastatic) cell lines to conduct mechanism studies by treating them with 10-40 μmol/L resveratrol and its analogs for 24 hours. In mouse subcutaneous and orthotopic xenograft models, 50 mg/kg was administered intragastrically for 5-7 weeks to evaluate its in vivo anti-tumor effect. The study found that resveratrol inhibits the proliferation, metastasis, and induces apoptosis of prostate cancer cells by downregulating the expression of epigenetic regulator MTA1 and oncogenic miRNAs (such as miR-21, miR-17-92 cluster).

 

4.2 Specifically Regulate the miR-17-PTEN Signaling Axis

The overexpression of oncogenic microRNAs (miRNAs), such as the miR-17 family, is closely related to the silencing of tumor suppressor genes such as PTEN [40]. Kumar et al. [41] revealed that resveratrol and its methylated analog pterostilbene can specifically reduce the expression levels of oncogenic miR-17 family members (such as miR-17-5p), relieve their post-transcriptional inhibition of the tumor suppressor gene PTEN, thereby restoring the regulatory function of PTEN-mediated PI3K/Akt signaling pathway, helping to inhibit tumor cell proliferation and promote their apoptosis.

The above studies provide a certain theoretical basis for the intervention strategy of resveratrol in epigenetics and its application in precision medicine, and highlight the importance of developing resveratrol analogs with high bioavailability in clinical transformation. For herbal developers, the epigenetic regulatory potential of resveratrol, together with Cistanche Tubulosa's scientifically proven benefits in improving sperm health and hormonal balance [24], underscores the value of natural herbs in addressing prostate issues from both genetic and physiological levels.

 

5 Targeting Mitochondrial Function

Mitochondria are multifunctional organelles that play key roles in various metabolic and cellular functions. It is generally believed that sound mitochondrial function is crucial for tumor cell proliferation; however, the mechanism by which changes in mitochondrial function affect cancer progression is not yet fully clear. Studies have found that mitochondria can promote tumor occurrence and development by regulating anabolism, intracellular signal transduction, and the tumor microenvironment. Therefore, mitochondrial metabolic reprogramming has become a key focus in cancer research [42].

 

5.1 Inhibit Mitochondrial Membrane Lipid Remodeling and COX Activity

Cardiolipin is the core phospholipid of the inner mitochondrial membrane, and its reduction may disrupt the stability of electron transport chain complexes (such as COX). Zichri et al. [43] conducted studies on a model system composed of human prostate cancer PC-3 cells and their controls using LC-MS lipidomics analysis, electron paramagnetic resonance spectroscopy, and enzyme activity assay techniques. They found that changes in the mitochondrial membrane lipid composition of prostate cancer cells include a significant increase in cardiolipin content, a decrease in zwitterionic lipids, and an increase in fatty acyl chain unsaturation, leading to enhanced membrane fluidity and reduced surface charge; in addition, the COX activity of cancer cells was significantly higher than that of normal cells, which may be related to the increase in cardiolipin content. After treating prostate cancer cells with resveratrol, the results showed that resveratrol accumulates in mitochondria and significantly inhibits the COX activity of cancer cells. This study systematically clarifies the potential mechanism of mitochondrial membrane lipid remodeling in abnormal energy metabolism of cancer cells, providing a theoretical basis for the development of precision anti-cancer drugs targeting mitochondrial membranes.

 

5.2 Induce Mitochondria-Mediated Caspase-Independent Apoptosis

Kumar et al. [44] used transgenic mouse prostate adenocarcinoma cells TRAMP-C1, TRAMP-C2, and TRAMP-C3 as models to explore the mechanism of apoptosis induction by treating cells with 50 and 100 μmol/L resveratrol for 16 hours. It was found that resveratrol can disrupt the mitochondrial membrane potential (Δψm), thereby leading to the upregulation of the pro-apoptotic protein Bax and the downregulation of the anti-apoptotic protein Bcl2. This finding indicates that resveratrol can trigger mitochondria-dependent apoptotic signals. In addition, the results showed that although resveratrol induces typical apoptotic phenomena such as DNA fragmentation, the Caspase-3 inhibitor Z-VAD-FMK failed to reverse cell death, confirming that its effect is independent of the Caspase pathway, suggesting the existence of a new non-classical apoptotic pathway. In addition, resveratrol-induced low-molecular-weight fragmentation of genomic DNA was observed in all three subtypes of TRAMP-C1/C2/C3, indicating that its mechanism of action is universal. This study reveals that resveratrol promotes apoptosis of prostate cancer cells through a mitochondria-mediated Caspase-independent pathway.

These studies reveal that resveratrol exerts anti-cancer effects by targeting the mitochondrial function of prostate cancer cells. In future studies, real-time imaging technology combined with metabolomics can be used to analyze the dynamic effects of resveratrol on mitochondrial oxidative phosphorylation, ROS production, and lipid metabolism networks. For herbal therapy developers, the combination of resveratrol's mitochondrial targeting and Cistanche Tubulosa's antioxidant protection (by boosting SOD and catalase activities [24]) can provide a dual protective mechanism against prostate cell damage, enhancing the potential of herbal-based treatments.

 

6 Conclusion

As natural active compounds with multi-target characteristics, resveratrol and Cistanche Tubulosa each demonstrate multi-dimensional, synergistic regulatory mechanisms in inhibiting prostate cancer progression and improving prostate health. Resveratrol inhibits tumor cell proliferation, promotes apoptosis and autophagy, blocks migration and invasion, and involves interventions at the epigenetic and metabolic levels, highlighting its potential in chemoprevention and adjuvant therapy. Cistanche Tubulosa, known as the "Ginseng of the Desert," complements these effects by reducing inflammation and oxidative stress, improving urinary function, regulating hormonal balance, and enhancing sperm health-all supported by clinical trials and PubMed studies [24].

Currently, the translational application of these compounds still faces challenges such as low bioavailability, unstable in vivo effects, and complex mechanisms. For developers interested in herbal therapies for prostatitis and prostate cancer, future research can focus on the following directions: systematically revealing the dynamic regulatory relationships between resveratrol/Cistanche Tubulosa and metabolic, epigenetic, and immune micro-networks in prostate cancer through experiments; striving to improve the stability and targeted delivery efficiency of these herbal ingredients; thoroughly verifying their synergistic effects with existing treatment strategies; promoting the transformation of relevant basic research results into clinical applications and conducting high-quality clinical trials.

Through interdisciplinary integration and technological integration, resveratrol and Cistanche Tubulosa are expected to open up new ideas and provide new methods for the precision treatment of prostate cancer, answering the key question "What is the best herbal for prostate?" with scientific evidence and practical potential. These natural herbs not only offer a safer alternative to traditional remedies (with no harmful side effects and ease of long-term use [24]) but also address the root causes of prostate issues rather than just alleviating symptoms, making them valuable candidates for the development of next-generation herbal prostate therapies.

 

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