Disulfiram alone regulates the radiosensitivity of lung cancer through NF-κB pathway and regulates the immune microenvironment after radiotherapy by targeting PD-L1 through c-Myc

Lung cancer is the leading cause of cancer-related mortality in the world. Metastasis is the leading cause of death and an enormous therapeutic challenge for non-small cell lung cancer. Radiation therapy is a treatment of unresectable lung cancer and may palliate the symptoms, locally control tumor growth, and provide higher survival rate. Yet, the molecular mechanism of radio-sensitivity in lung cancer still remains unclear. Two lung adenocarcinoma cell lines (CL1-0 and CL1-5) with different metastatic potentials were irradiated with 10Gy γ-radiation. CL1-5 showed more radio-sensitive than CL1-0 since the surviving fractions in CL1-5 were ten times lower than CL1-0 by clonogenic assays. Gene expression profiles were also analyzed using microarray in order to better understand differential gene expression between these two cell lines after radiation. Total RNAs for whole genome gene expression analysis were extracted from these two cell lines at 0, 1, 4, 9 and 24 h after 10Gy irradiation. Gene Set Enrichment Analysis (GSEA) revealed the pathways of death, cytokine, cell adhesion, IL1R, NF-κB, and TNFR had differences between these two cell lines following 10Gy radiation exposure. Tumor necrosis factor (TNF) cytokine family and interleukin-1 (IL1) was up-regulated in CL1-5 after irradiation. The expression level of TNF-α validated by quantitative real time PCR was increased in CL1-5 and was not detected in CL1-0. Therefore, TNF-α released into the culture medium in CL1-5 after irradiation might be a death signal to activate the NF-κB pathway and cause more cell death. Moreover, the different responses of NF-κB-related pathways induced by radiation between the two cell lines might contribute the different survival. These cytokine modulations might provide promising targets for further investigation regarding radiation treatment in lung cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1326.

Radiotherapy is a critical approach for the comprehensive treatment of non-small cell lung cancer. Deep understanding of the individualized radiosensitivity of lung cancer patients plays a pivotal role in the selection of radiotherapy dosage and regime and establishment of comprehensive therapeutic strategies. Currently, multiple researchers have identified a variety of biomarkers in predicting the radiosensitivity of lung cancer patients. In this article, research progress on the biomarkers in predicting radiosensitivity of non-small cell lung cancer was reviewed. Key words: Lung neoplasm; Radiosensitivity; Biomarker

Identifying new targets for overcoming radioresistance is crucial for improving the efficacy of lung cancer radiotherapy, given that tumor cell resistance is a leading cause of treatment failure. Recent research has spotlighted the significance of Musashi2 (MSI2) in cancer biology. In this study, we first demonstrated that MSI2 plays a key function in regulating the radiosensitivity of lung cancer. The expression of MSI2 is negatively correlated with overall survival in cancer patients, and the knockdown of MSI2 inhibits tumorigenesis and increases radiosensitivity of lung cancer cells. Cellular radiosensitivity, which is closely linked to DNA damage, is influenced by MSI2 interaction with ataxia telangiectasia mutated and Rad3-related kinase (ATR) and checkpoint kinase 1 (CHK1) post-irradiation; moreover, knockdown of MSI2 inhibits the ATR-mediated DNA damage response pathway. RNA-binding motif protein 17 (RBM17), which is implicated in DNA damage repair, exhibits increased interaction with MSI2 post-irradiation. We found that knockdown of RBM17 disrupted the interaction between MSI2 and ATR post-irradiation and increased the radiosensitivity of lung cancer cells. Furthermore, we revealed the potential mechanism of MSI2 recruitment into the nucleus with the assistance of RBM17 to activate ATR to promote radioresistance. This study provides novel insights into the potential application of MSI2 as a new target in lung cancer radiotherapy.

Enhanced Radiosensitivity of Esophagus Cancer through Loss of ADAR1 and Cell Apoptosis via NF-kB Signaling Pathway

Dezocine is an opioid analgesic that can affect the immune system. Here, we explored the synergy of high concentration of Dezocine and programmed death-ligand 1 (PD-L1) with regards to immune escape and glucose metabolism in lung cancer (LC). PD-L1 level in human LC cell lines was determined and the influence of Dezocine at different concentrations for the proliferation of LC cells was identified. Next, LC cells were transfected to alter PD-L1 level, and exposed to Dezocine at 8 μg/mL to explore their effects on cell proliferation, production of interferon-γ (IFN-γ), contents of glucose, lactate, and NADPH/NADP+, and activation of the nuclear factor-κB (NF-κB) pathway. PD-L1 level was increased in LC cells and Dezocine (8 μg/mL) impaired the proliferation of LC cells. Down-regulating PD-L1 inhibited cell proliferation, enhanced production of IFN-γ, and reduced the contents of glucose, lactate, and NADPH/NADP+, while up-regulating PD-L1 caused the opposite results. Dezocine (8 μg/mL) induced immune escape and glucose metabolism in LC, and Dezocine-induced effects were reversed by down-regulating PD-L1. Dezocine (8 μg/mL) up-regulated PD-L1 by activating the NF-κB pathway. Dezocine at 8 μg/mL promotes immune escape and glucose metabolism in LC through up-regulating PD-L1 and activating the NF-κB pathway.

Transforming growth factor-activated kinase 1 (TAK1)-binding protein 3 (TAB3) is essential for the activation of the nuclear factor kappa B (NF-κB) pathway and has important roles in cell survival. However, the contribution of TAB3 to non-small cell lung cancer (NSCLC) remains elusive. In the present study, Western blotting and immunohistochemistry assays demonstrated that TAB3 expression was frequently increased in NSCLC tissues and cells. In addition, chi-square test and Kaplan-Meier analysis revealed that upregulation of TAB3 expression correlated with a more invasive tumor phenotype and poor prognosis. In addition, a series of experiments, including serum starvation-refeeding experiment and TAB3-siRNA transfection assay, showed that TAB3 expression promoted NSCLC cell proliferation. Furthermore, the effect of TAB3 expression on the sensitivity to cis-diamminedichloroplatinum (CDDP) and possible signaling transduction pathways was investigated. When the expression of TAB3 was inhibited by siRNA transfection, the sensitivity to CDDP was enhanced. Moreover, it showed that downregulation of TAB3 enhanced CDDP-induced A549 cell apoptosis through the inhibition of the NF-κB pathway. These results suggest that TAB3 plays a critical role in NSCLC progression and chemoresistance and that TAB3 depletion may be a promising approach to lung cancer therapy.

The crosstalk between α-irradiated Beas-2B cells and its bystander U937 cells through MAPK and NF-κB signaling pathways

Cancer remains a leading cause of death, and current therapeutic options designed to slow the progression of cancer or eradicate cancer cells are often limited by drug resistance, inefficacy, or adverse effects. The Nuclear Factor Kappa B (NF-κB) pathway is a central regulator of inflammation and immune responses, and its dysregulation contributes to cancer development and progression. This review provides an overview of the role of the NF-κB pathway in tumor development and progression and discusses the potential of targeting specific modulators of the pathway for cancer drug discovery, specifically cancers that have the highest prevalence, such as breast, colorectal, lung, melanoma, and prostate cancers. While NF-κB inhibitors show promise, particularly in hematologic malignancies, challenges remain in translating these findings to solid tumors due to pathway complexity and its essential role in normal immunity.

Cell Signaling/Signal Transduction

We have previously reported that genistein, the major bioactive isoflavone of soybeans, acts as a radiosensitizer for prostate cancer (PCa) both in vitro and in orthotopic mouse models in vivo. However, pure genistein promoted increased spontaneous metastasis to regional lymph nodes, when administered alone in vivo. In contrast to genistein, a natural formulation of soy isoflavones mixture (43% genistein, 21% daidzein and 2% glycitein) did not cause increased metastasis, but like genistein, the soy mixture potentiated the therapeutic effect of radiation. We have now tested whether daidzein, the second main isoflavone of soybeans, could negate genistein-induced lymph node metastasis in PC-3 orthotopic prostate tumors in nude mice. Mice were treated with purified daidzein, or genistein or both combined; and also in conjunction with prostate tumor irradiation. Either daidzein or genistein alone or both combined caused the same extent of prostate tumor growth inhibition than the soy mixture. Treatment with daidzein alone did not increase metastasis to regional lymph nodes but still acted as a potent radiosensitizer for prostate tumors. Treatment with both genistein and daidzein combined did not cause an increase in lymph node metastasis, which is consistent with the findings obtained with the soy mixture. These studies show that pure daidzein could be the component of the soy isoflavones mixture that protects against genistein-induced lymph node metastasis. In order to gain further mechanistic insight on how daidzein negates the adverse effects of genistein, we compared the effects of each isoflavone and the soy mixture in vitro using PC-3 (androgen receptor AR-) and C4-2B (AR+) androgen independent PCa cell lines. We found that daidzein inhibited tumor cell growth and synergized with radiation in clonogenic assays but at doses higher than genistein or the soy mixture. We have previously demonstrated that genistein or the soy mixture cause tumor cell apoptosis and enhance radiation killing by inhibiting cell survival pathways activated by radiation including the repair enzyme/redox activator APE1/Ref-1 and the transcription factors HIF-1α and NF-κB. Compared to the effect of genistein or the soy mixture, daidzein treatment caused milder effects on PARP cleavage, inhibition of expression of APE1/Ref-1 and HIF-1α; and lower inhibition of HIF-1α and NF-κB DNA binding activities. These effects were reproduced in both PC-3 and C4-2B cell lines. Based on these results, we conclude that APE1/Ref-1, NF-κB and HIF-1α molecular pathways were also affected by daidzein, although to a lesser extent than genistein and soy mixture, in both AR+ and AR- PCa cancer cell lines, suggesting that this is an AR-independent mechanism. The protective effect of daidzein against lymph node metastasis induced by genistein appears to be independent of HIF-1α and NF-κB pathways, and remains to be clarified. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5541.

Radiotherapy is one of the cornerstone treatments for endometrial cancer and has successfully diminished the risk of local recurrences after surgery. However, a considerable percentage of patients suffers tumor relapse due to radioresistance mechanisms. Knowledge about the molecular determinants that confer radioresistance or radiosensitivity in endometrial cancer is still partial, as opposed to other cancers. In this review, we have highlighted different central cellular signaling pathways and processes that are known to modulate response to radiotherapy in endometrial cancer such as PI3K/AKT, MAPK and NF-κB pathways, growth factor receptor signaling, DNA damage repair mechanisms and the immune system. Moreover, we have listed different clinical trials employing targeted therapies against some of the aforementioned signaling pathways and members with radiotherapy. Finally, we have identified the latest advances in radiotherapy that have started being utilized in endometrial cancer, which include modern radiotherapy and radiogenomics. New molecular and genetic studies in association with the analysis of radiation responses in endometrial cancer will assist clinicians in taking suitable decisions for each individual patient and pave the path for personalized radiotherapy.

Rationale: Radioresistance remains the major cause of local relapse and distant metastasis in lung cancer. However, the underlying molecular mechanisms remain poorly defined. This study aimed to investigate the role and regulatory mechanism of Cyclin K in lung cancer radioresistance.Methods: Expression levels of Cyclin K were measured by immunohistochemistry in human lung cancer tissues and adjacent normal lung tissues. Cell growth and proliferation, neutral comet and foci formation assays, G2/M checkpoint and a xenograft mouse model were used for functional analyses. Gene expression was examined by RNA sequencing and quantitative real-time PCR. Protein-protein interaction was assessed by immunoprecipitation and GST pull-down assays.Results: We report that Cyclin K is frequently overexpressed and correlates with poor prognosis in lung cancer patients. Functionally, we demonstrate that Cyclin K depletion results in reduced proliferation, defective G2/M checkpoint and enhanced radiosensitivity in lung cancer. Mechanistically, we reveal that Cyclin K interacts with and promotes the stabilization of β-catenin protein, thereby upregulating the expression of Cyclin D1. More importantly, we show that Cyclin D1 is the major effector that mediates the biological functions of Cyclin K in lung cancer.Conclusions: These findings suggest that Cyclin K positively modulates the β-catenin/Cyclin D1 axis to promote tumorigenesis and radioresistance in lung cancer, indicating that Cyclin K may represent a novel attractive biomarker for lung cancer radiotherapy.

USP28-mediated deubiquitination of FOXK1 activates the Hippo signaling pathway to regulate cell proliferation and radiosensitivity in lung cancer.

BackgroundAlthough immune checkpoint blockade (ICB) has been proven to achieve a persistent therapeutic response in various tumor types, only 20%–40% of patients benefit from this treatment. Radiotherapy (RT) can enhance...

Lung cancer (LC) is the most prevalent cancer with a poor prognosis. Semaphorin4A (Sema4A) is important in many physiological and pathological processes. This study aimed to explore the role and mechanism of Sema4A in LC. Firstly, Sema4A expression was analyzed by the available dataset and detected in human normal bronchial epithelial cell line (HBE) and LC cell line (NCI-H460). Then, LC cells were transfected with Sema4A siRNA, and the cells were stimulated by PlexinB1, PlexinB2, PlexinD1 blocking antibodies, IgG antibody, BAY 11-7082 (an inhibitor for NF-κB pathway) and Sema4A-Fc protein, alone or in combination. After transfection, PlexinB1 mRNA expression was analyzed. Next, the biological functions, including proliferative, migratory, invasive abilities and viability of the cells were detected by colony formation, scratch, Transwell and MTT assays, respectively. NF-κB, Stat3 and MAPK protein expressions were determined by western blot. Furthermore, the secretion of IL-6 in LC cells was tested by ELISA. Sema4A was highly expressed in LC tissues and cells, could activate the NF-κB pathway and upregulate PlexinB1 mRNA expression. Furthermore, we observed that Sema4A knockdown suppressed the biological functions of NCI-H460 cells, while Sema4A-Fc protein reversed the situation. However, Sema4A-induced biological functions and activation in the NF-κB pathway were inhibited by PlexinB1 blocking antibody. Consistently, Sema4A promoted IL-6 production, which was down-regulated by PlexinB1 blocking antibody and BAY 11-7082. Sema4A may facilitate LC development via the activation of the NF-κB pathway mediated by PlexinB1, suggesting that Sema4A would be a novel therapeutic target for LC treatment.