Elucidating the Impact of Hypericum alpestre Extract and L-NAME on the PI3K/Akt Signaling Pathway in A549 Lung Adenocarcinoma Cells

Javrushyan, H.; Ginovyan, M.; Petrosyan, G.; Qocharyan, M.; Harutyunyan, T.; Gevorgyan, S.; Karabekian, Z.; Maloyan, A.; Avtandilyan, N.

doi:10.1101/2024.05.02.592167

bioRxiv · 10.1101/2024.05.02.592167

Elucidating the Impact of Hypericum alpestre Extract and L-NAME on the PI3K/Akt Signaling Pathway in A549 Lung Adenocarcinoma Cells

Javrushyan, H.·Ginovyan, M.·Petrosyan, G.·Qocharyan, M.·Harutyunyan, T.·Gevorgyan, S.·Karabekian, Z.·Maloyan, A.·Avtandilyan, N.

View original source ↗Export metadata ↓

Abstract

Plants within the Hypericaceae family have been traditionally used for their medicinal properties, showcasing a wide range of effects such as antibacterial, antiviral, and antioxidant qualities. Hypericum alpestre (HA) extracts have exhibited significant cytotoxicity against various cancer cell lines. The phenolic compounds found in HA extracts have attracted attention for their potential in cancer prevention. L-NAME, known for its ability to inhibit nitric oxide synthase (NOS) activity, has emerged as a promising approach in cancer therapy. However, the precise molecular mechanisms underlying the anticancer effects of HA and L-NAME remain unclear. This study aims to clarify the impact of HA and L-NAME on the PI3K/Akt signaling pathway in A549 lung adenocarcinoma cells, with a specific focus on TNFa/COX-2 and VEGFa/MMP-2 pathways. In silico analysis, they identified the compounds with the highest affinity for PI3K/Akt, a finding validated by subsequent in vitro experiments. Furthermore, the combination of herbs and L-NAME exhibited superior efficacy compared to the herb and 5-FU combination, as evidenced by the promotion of apoptosis. Both the herb alone and the combination of the herb with L-NAME demonstrated inhibitory effects on the TNFa/COX-2 and VEGFa/MMP-2 pathways. This therapeutic approach is hypothesized to operate through the PI3k/Akt cell signaling pathway. A better understanding of the interaction between HA polyphenols and PI3K/Akt signaling could pave the way for novel therapeutic strategies against cancer, including drug-resistant tumors.

Explore related subjects

cancer biology Preprint

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Javrushyan, H., Ginovyan, M., Petrosyan, G., Qocharyan, M., Harutyunyan, T., Gevorgyan, S., Karabekian, Z., Maloyan, A., Avtandilyan, N.. 2024-05-04. Elucidating the Impact of Hypericum alpestre Extract and L-NAME on the PI3K/Akt Signaling Pathway in A549 Lung Adenocarcinoma Cells. https://doi.org/10.1101/2024.05.02.592167

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

Tissue resident CD4+ memory T-cells mark response to immune checkpoint inhibition in high-grade glioma

Background: Immune checkpoint inhibitors (ICI) are efficacious in many solid tumors, but response in glioma is restricted to a small subgroup. The determinants of response and resistance to ICI remain poorly understood. Methods: Here we exploit a syngeneic hypermutated high-grade glioma model with dichotomous response to combined PD-1 and CTLA-4 inhibition to unravel determinants of tumor-infiltrating T-cells driving response. Tumor-infiltrating T-cells from ICI-responsive and non-responsive tumors were analyzed by single-cell RNA and T-cell receptor sequencing and tumor-reactive T-cell receptor clonotypes were functionally validated to characterize their transcriptional phenotypes. We verify our findings in IDH1 wildtype glioblastoma patients treated with neoadjuvant pembrolizumab. Results: ICI response was associated with intratumoral clonal expansion of tumor-reactive cytotoxic T-cells and increased infiltration of CXCR6+ CD4+ tissue resident memory T-cells (Trm). CD4 stem-like memory T-cells in responding tumors demonstrated elevated interferon responses, following trajectories toward clonally expanded Trm, versus trajectories toward exhaustion in non-responsive tumors. In responsive tumors, CD4+ Trm interacted with infiltrating CXCR3+ tumor-reactive and clonally expanded, yet transcriptionally versatile cytotoxic T-cells. Probing the post neoadjuvant ICI high-grade glioma patient tissue dataset, we confirmed increased CXCR6 expression in CD4+ T cells and the association of CD4+ Trm with prolonged overall survival. Conclusion: These findings identify CD4 tissue-resident memory T-cells as determinants of ICI response in IDH1 wildtype high-grade glioma and warrant their further investigation to improve immunotherapy outcomes.

cancer biology↗

bioRxiv2026

Circadian gene-network distortion in high-risk neuroblastoma across multiple biological reference contexts

Background: The circadian clock regulates cellular homeostasis, and its disruption has been implicated in aggressive neuroblastoma, particularly in tumours harbouring MYCN- amplification. However, it remains unclear whether alterations are restricted to individual clock genes or extend to circadian gene network coordination. We therefore examined circadian clock network disruption in adverse neuroblastoma across multiple biological contexts. Methods: We estimated circadian gene network dysregulation using Delta-CCD in tumours from two neuroblastoma cohorts (SEQC n=498 and Kocak n=649), comparing clinical features associated with outcome across canonical, adrenal-tissue matched, and developmental references. Robustness was assessed by cross-cohort meta-analysis and leave-one-gene-out analyses. Cox proportional hazards models adjusted for clinical covariates assessed association between individual clock gene expression patient outcome. Results: Delta-CCD was highest in tumours classified as high-risk (study-specific definition) across reference contexts in both cohorts. MYCN-amplified tumours showed a more reference-dependent pattern, strongest in adrenal context, while stage 4 tumours showed a similar but weaker pattern. Additional analyses supported the high-risk signal as a distributed network-level alteration rather than a single-gene phenomenon. Conclusions: High-risk neuroblastoma is characterised by robust disruption of coordinated clock gene network organisation across canonical and tissue-matched references, extending beyond individual clock genes. The extent of circadian dysregulation depends on the reference state used.

cancer biology↗

bioRxiv2026

BAP1 loss and PRAME expression converge to remodel the tumor-immune ecosystem during uveal melanoma progression

Uveal melanoma (UM) is characterized by a small number of recurrent genetic alterations that determine metastatic propensity. BAP1 loss and PRAME expression define the dominant prognostic axes in UM, yet how they promote malignant progression remains unclear. We profiled 190,535 cells from normal uvea, uveal nevus, primary and metastatic UM using single-cell transcriptomics, T cell receptor sequencing, spatial transcriptomics and isogenic perturbation models. Normal melanocytes, nevus cells and UM cells formed a transcriptional continuum marked by loss of differentiation and emergence of neural crest-like, stress-responsive, hypoxic-glycolytic and immune-interacting states. BAP1 loss and PRAME expression imposed distinct but convergent immunoregulatory programs, inducing interferon and TNF-NFkB signaling and MHC-I expression, with HLA-E showing the strongest response. These alterations were accompanied by macrophage and CD8+ T cell remodeling. PRAME-enriched tumor regions formed spatially organized niches enriched for macrophages and plasma cells. These findings define BAP1 loss and PRAME expression as distinct but convergent axes of tumor-immune coevolution and nominate HLA-E as a candidate mediator of immune resistance.

cancer biology↗