Adenocarcinoma Cell Lung Cancer Research Articles

Anti-cancer activity of NTAX-44 (bioprocessed arsenic trioxide) on lung cancer.

e20573 Background: In contemporary allopathic medicine, intravenous administration of Arsenic trioxide is extensively investigated for its diverse applications in cancer treatment. However, the associated adverse effects demand exploration of alternative formulations. In present study, we propose the patented green technology-based compound “oral Arsenic Trioxide” (NTAX-44), in its unique form that can be administered orally, thereby reduces cost of treatment. This product presenting itself as a promising candidate for cancer therapy. Despite the ethnopharmacological uses of arsenicals in medicine, comprehensive understanding of their safety and efficacy remains limited. Thus, present study is proposed to investigate anticancer potential of NTAX-44 in lung adenocarcinoma cancer cell line A549. Methods: The effect of oral NTAX-44 was assessed on the viability of cell lines grown as 2D monolayers by MTS assay and 3D multicellular spheroid (MCS) model. Anti-metastatic potential of NTAX-44 was studied by using migration and colony formation assay while, expression of PD-L1 in A549 cell line treated with Arsenic Trioxide was checked by flow cytometry. The anti-metastatic capacity and cytocompatibility testing of Arsenic Trioxide was evaluated by in ovo chick embryo yolk sac membrane (YSM) angiogenesis assay and MTS assay using PBMCs respectively. Results: NTAX-44 is found to be effective to inhibit viability of A549 cell line (IC50 21.47±1.7 μg/ml for 48 hr and 14.65±0.92 for 72 hr of treatment); exhibiting its anti-proliferative activity. The A549 spheroids showed comparatively more resistance with loss of viability at higher concentration (50 and 100 µg/ml). Statistically significant decrease in scratch repair rate was observed at 24 hr and 72 hr of treatment and showing anti-migratory activity in A549 cells. In colony formation assay, complete inhibition of the colonization of A549 cells was observed after 48 hr of treatment; the survival fraction identified as 0%. NTAX-44 at 25µg/ml concentration inhibited PD-L1 expression in A549 cells, suggesting its potential to decrease expression of PD-L1 on tumor cells resulting in reversal of immune suppressive tumor microenvironment. Treatment of YSM with NTAX-44 resulted in approximately 40% inhibition in formation of quaternary blood vessels comparable to Bevacizumab (Avastin®). This significant anti-angiogenic potential is suggestive of its in vivo anti-metastatic effect. Notably, cytocompatibility testing of NTAX-44 at concentrations up to 200μg/ml was well-tolerated in PBMC cells emphasizing its favourable biocompatibility profile, providing crucial insights into its safety. Conclusions: Inhibitory effect of NTAX-44 on cell proliferation serves as a noteworthy attribute in context of its potential therapeutic utility against lung cancer. Its cytocompatibility further indicates application of arsenic trioxide as a candidate for clinical application.

Synthesis, crystal structure, Hirshfeld, DFT, molecular docking, dynamics studies, and anti-cancer activity of 1-substituted-2-(4-(diethylamino)-2-hydroxyphenyl)- 1H-benzo[d]imidazole-5-ethyl carboxylates

In our present work, a novel series of 1-substituted-2–2-(4-(diethylamino)-2-hydroxyphenyl)- 1H-benzo[d]imidazole-5-ethyl carboxylates were synthesized by an efficient “one-pot” nitro reductive cyclization method using sodium dithionite is reported as potential anticancer agents. IR, 1H & 13C NMR and Mass spectral data confirmed the synthesized compounds' structure. Compound 6a and 6j were confirmed by their single crystal XRD studies. The Hirshfeld surfaces computational method was applied to visualize and analyse the intermolecular interactions within the crystal structure. All the synthesized compounds were assessed for in vitro anticancer activity towards human lung adenocarcinoma A549 cells and breast cancer MCF-7 cell lines. Compound 6j showed a promising activity with an IC50 value 42.7 μg/mL and 89.63 μg/mL against A549 and MCF-7 cell lines respectively. The potent compounds were assessed for cytotoxicity study on human embryonic kidney HEK 293 cell line and found to be non-toxic. Also, apoptosis study indicated compound 6j induced early apoptosis in the A549 lung adenocarcinoma and MCF-7 breast cancer cells. Molecular docking and dynamic simulation studies were carried out on human estrogen receptor protein (PDB: 3ERT).

Abstract 3147: DXC025, a novel anti-MUC1/EGFR bispecific antibody-tubulysin conjugate with a function linker, exhibits potent anti-tumor efficacy

Abstract Antibody-drug conjugates (ADCs) have become a promised drug class in cancer therapy. Armed with three main components-an antibody, a linker molecule, and a cytotoxic agent (“payload”), plus sites of the antibody conjugation and the distribution of tumor-associate antigens, nowadays ADCs have proved the unique ability of the precision of targeted therapy, a great leap forward from traditional chemotherapeutic approaches that cause widespread effects without specificity. Both epidermal growth factor receptor (EGFR) and mucin 1 (MUC1) are tumor-associated antigens (TAA) that are co-expressed in many solid tumors, such as, colon cancer, non-small cell lung cancer (NSCLC), esophageal cancer, epidermal cancer, and pancreatic cancer. In particular, MUC1, a glycoprotein essential for the formation of the epithelial mucous barrier, is hypoglycosylated and dimerizes with EGFR, a potent oncoprotein, in transformed cells. For our anti-Muc1 ADC clinical program, for treatment of colon and pancreatic cancers, it was found out that the highly glycosylated transmembrane Muc1 had much more turning over rates on the luminal surface of epithelial cells. Here, we first designed and generated bispecific Fab/scFv antibodies targeting both MUC1 and EGFR, using the knob-into-hole technology. The addition of the affinity- and internalization-optimized anti-EGFR arm to the other single Muc1 arm was aimed to broaden tumor selectivity and to reduce on-target toxicity, for the treatment of gastrointestinal tumors in comparison to individual full IgG1-ADC, either EGFR or Muc1-ADC. Indeed, many of generated anti- MUC1/EGFR bispecific antibodies (BsAbs) showed strong binding affinity in MUC1modetateEGFRmodetate cells, in addition to both MUC1 and EGFR highly expressed cells. Internalization assays demonstrated that the BsAbs were endocytosed in tumor cells co-expressing EGFR and MUC1 more efficiently than monoclonal antibodies targeting MUC1. The selected BsAbs were subsequently conjugated with tubulysin B analogs with varieties of Tubulysin B analog payloads and function peptidyl spacer/linkers to generate bispecific ADC (DXC025) candidates. DXC025 candidates exhibited very potent cytotoxicity in vitro against epidermal cancer, lung adenocarcinoma, and pancreatic cancer cell lines with IC50 of single or tens digital pM. In in vivo cell-derived xenograft (CDX) models, DXC025 candidates demonstrated much superior efficacies of tumor growth inhibition in comparison with parental (monoclonal), either Muc1- or EGFR- ADC for the treatment of gastrointestinal tumors. These results indicated that the MUC1/EGFR-ADC (DXC025) would be a promising ADC candidate for targeted treatment of either Muc1- or EGFR-expressing gastrointestinal tumors. Citation Format: Xingyan Jiang, Junxiang Jia, Huihui Guo, Xiangfei Kong, Yongfang Xu, Sishi Ye, Yong Du, Zhicang Ye, Qikuang Cen, Lingli Zhang, Yongxiang Chen, Gaituo Chen, Lu Bai, Yunxia Zheng, Wei Zheng, Jun Zheng, Juan Wang, Wenjun Li, Yuanyuan Huang, Linyao Zhao, Yifang Xu, Mengmeng Liu, Binbin Chen, Meng Dai, Miaomiao Chen, Zhixiang Guo, Qingliang Yang, Robert Y. Zhao. DXC025, a novel anti-MUC1/EGFR bispecific antibody-tubulysin conjugate with a function linker, exhibits potent anti-tumor efficacy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3147.

SOX2 Expression Does Not Guarantee Cancer Stem Cell-like Characteristics in Lung Adenocarcinoma.

Effectively targeting cancer stemness is essential for successful cancer therapy. Recent studies have revealed that SOX2, a pluripotent stem cell factor, significantly contributes to cancer stem cell (CSC)-like characteristics closely associated with cancer malignancy. However, its contradictory impact on patient survival in specific cancer types, including lung adenocarcinoma (LUAD), underscores the need for more comprehensive research to clarify its functional effect on cancer stemness. In this study, we demonstrate that SOX2 is not universally required for the regulation of CSC-like properties in LUAD. We generated SOX2 knockouts in A549, H358, and HCC827 LUAD cells using the CRISPR/Cas9 system. Our results reveal unchanged CSC characteristics, including sustained proliferation, tumor sphere formation, invasion, migration, and therapy resistance, compared to normal cells. Conversely, SOX2 knockdown using conditional shRNA targeting SOX2, significantly reduced CSC traits. However, these loss-of-function effects were not rescued by SOX2 resistant to shRNA, underscoring the potential for SOX2 protein level-independent results in prior siRNA- or shRNA-based research. Ultimately, our findings demonstrate that SOX2 is not absolutely essential in LUAD cancer cells. This emphasizes the necessity of considering cancer subtype-dependent and context-dependent factors when targeting SOX2 overexpression as a potential therapeutic vulnerability in diverse cancers.

Neutrophil infiltration associated genes on the prognosis and tumor immune microenvironment of lung adenocarcinoma.

The neutrophils exhibit both anti-tumor and pro-tumor effects in cancers. The correlation between neutrophils and tumor development in lung adenocarcinoma (LUAD) is still uncertain, possibly due to a lack of specific neutrophil infiltration evaluation methods. In this study, we identified 30 hub genes that were significantly associated with neutrophil infiltration in LUAD through data mining, survival analysis, and multiple tumor-infiltrating immune cells (TICs) analysis, including TIMER, CIBERSORT, QUANTISEQ, XCELL, and MCPCOUNTER. Consensus clustering analysis showed that these 30 hub genes were correlated with clinical features in LUAD. We further developed a neutrophil scoring system based on these hub genes. The neutrophil score was significantly correlated with prognosis and tumor immune microenvironment (TIME) in LUAD. It was also positively associated with PD-L1 expression and negatively associated with tumor mutational burden (TMB). When combined with the neutrophil score, the predictive capacity of PD-L1 and TMB for prognosis was significantly improved. Thus, the 30 hub genes might play an essential role in the interaction of neutrophils and LUAD, and the neutrophil scoring system might effectually assess the infiltration of neutrophils. Furthermore, we verified the expression of these 30 genes in the LUAD tumor tissues collected from our department. We further found that overexpressed TNFAIP6 and TLR6 and downregulated P2RY13, SCARF1, DPEP2, PRAM1, CYP27A1, CFP, GPX3, and NCF1 in LUAD tissue might be potentially associated with neutrophils pro-tumor effects. The following in vitro experiments demonstrated that TNFAIP6 and TLR6 were significantly overexpressed, and P2RY13 and CYP27A1 were significantly downregulated in LUAD cell lines, compared to BEAS-2B cells. Knocking down TNFAIP6 in A549 and PC9 resulted in the upregulation of FAS, CCL3, and ICAM-1, and the downregulation of CCL2, CXCR4, and VEGF-A in neutrophils when co-culturing with the conditioned medium (CM) from LUAD cells. Knocking down TNFAIP6 in LUAD also led to an elevated early apoptosis rate of neutrophils. Therefore, overexpressed TNFAIP6 in LUAD cancer cells might lead to neutrophils "N2" polarization, which exhibited pro-tumor effects. Further research based on the genes identified in this pilot study might shed light on neutrophils' effects on LUAD in the future.

ARAP1 negatively regulates stress fibers formation and metastasis in lung adenocarcinoma via controlling Rho signaling

Small GTPases regulate multiple important cellular behaviors and their activities are strictly controlled by a mass of regulators. The dysfunction or abnormal expression of small GTPases or their regulators was frequently observed in various cancers. Here, we analyzed the expression and prognostic correlation of several GTPases and related regulators based on the TCGA database and found that Ankyrin Repeat and PH Domain 1 (ARAP1), a GTPase activating protein (GAP), is reduced in lung adenocarcinoma tissues compared to normal tissues and displays a positive correlation with overall survival (OS) and progression-free survival (PFS) of patients with lung adenocarcinoma. qPCR and western blot verified that ARAP1 is frequently downregulated in lung adenocarcinoma tumor tissues and cancer cells, and its downregulation might be mediated by epigenetic modification. Moreover, metastatic assays showed that overexpression of ARAP1 significantly inhibits metastasis of lung adenocarcinoma in vitro and in vivo. We further demonstrated that Rho signaling inhibition, mediated by RhoGAP activity of ARAP1, majorly contributes to suppressing migration and invasion of lung adenocarcinoma cancer cells via inhibiting stress fibers formation. In summary, this study indicates that ARAP1 may serve as a potential prognostic predictor and a metastatic suppressor in lung adenocarcinoma via its RhoGAP activity.

Novel circular RNA circ-0002727 regulates miR-144-3p/KIF14 pathway to promote lung adenocarcinoma progression.

Objective: Circular RNAs (circRNAs) have been shown to participate in various cancers via sponging miRNAs (microRNAs). However, their role in lung adenocarcinoma (LUAD) remains elusive. Methods: The transcriptome data and corresponding clinical information of lung adenocarcinoma samples were extracted from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) database. Differentially expressed circRNAs (DEcircRNAs), differentially expressed miRNAs (DEmiRNAs), and differentially expressed genes (DEgenes) were identified and further used to constructed a circRNA-associated competing endogenous RNA (ceRNA) network. Real-Time qPCR analysis was conducted to examine gene expression at transcriptional level. The regulatory mechanisms of circRNA-miRNA-gene were validated by dual-luciferase reporter array and RNA pull-down assay. Cell growth, migration and invasion were evaluated by CCK-8 assay, colony formation assay and transwell assay, respectively. Results: Based on public microarray data, we systematically constructed a circRNA-associated ceRNA network including 11 DEcircRNAs, 8 DEmiRNAs and 49 DEgenes. Among the ceRNA network, we found that circ-0002727 was a key regulatory and was further confirmed to be upregulated in LUAD cancer cells. Subsequently, we found that silencing of circ-0002727 significantly suppressed the LUAD cell proliferation, migration and invasion in vitro. Mechanistically, we showed that circ-0002727 could competitively bind miR-144-3p to enhance the KIF14 expression in LUAD cells. Rescue assays indicated that circ-0002727 could regulate LUAD cell proliferation through modulating miR-144-3p/KIF14 pathway. Besides, KIF14 expression level was positively correlated with TNM stage and metastasis, and patients with high KIF14 expression suffered poor prognosis. Conclusion: Taken together, our study revealed that circ-0002727 could act as a ceRNA to regulate LUAD progression via modulating miR-144-3p/KIF14 pathway, providing a potential therapeutic target for LUAD.

Determination of Structure and Cytotoxicity of Ten Undescribed Steroidal Glycosides from Allium cristophii × A. macleanii 'Globemaster'.

'Globemaster' is an ornamental hybrid cultivar whose parent plants are Allium cristophii and A. macleanii. The chemical constituents of 'Globemaster' bulbs have not yet been examined; thus, a systematic phytochemical investigation was undertaken herein. A series of chromatographic separations of the MeOH extract of 'Globemaster' bulbs afforded 27 steroidal glycosides (1-27), which are classified into 23 spirostanol glycosides (1-8 and 11-25), two furostanol glycosides (9 and 26), a pregnane glycoside (10), and a cholestane glycoside (27). The structures of the hitherto undescribed compounds (1-10) were determined from the two-dimensional NMR spectroscopic data and hydrolysis. The cytotoxicity of the isolated compounds (1-27) toward HL-60 human promyelocytic leukemia cells, A549 human adenocarcinoma lung cancer cells, and SBC-3 human small-cell lung cancer cells was evaluated. Compounds 8, 22, 23, 24, and 26 exhibited cytotoxicity toward all cell lines in a dose-dependent manner, with IC50 values in the 1.3-49 µM range.

Survival Rate of Lung Adenocarcinoma Patients Receiving EGFR - Tyrosine Kinase Inhibitor Targeted Therapy

Globally, lung cancer is by far the leading cause of death by cancer-which contribute to 2.094 million death-with the highest toll from cancer being 1.8 million. Currently, lung cancer therapy has developed from chemotherapy to targeted therapy, such as Epidermal Growth Factor Receptor-Tyrosine Kinase Inhibitor (EGFR-TKI). This study aimed to assess the survival rate of adenocarcinoma cell lung cancer patients who received EGFR-TKI therapy at the Pulmonary Clinic of Dr. Saiful Anwar General Hospital Malang. This study was a retrospective study using patient medical records between 2017 and 2020. The data were processed and analyzed using the Chi-Square test. The number of samples was 117 patients consisting of 63 patients receiving Gefitinib therapy, 36 patients receiving Afatinib therapy, and 18 patients receiving Erlotinib therapy. There were no significant differences between variables of age, sex, smoking history, stage, and exon mutations with 1-year survival. Gefitinib therapy has a higher average survival time than Afatinib and Erlotinib. However, the 1-year survival rate (YSR) was highest on Afatinib. The Middle Survival (MS) of the three regimens is almost the same, about 300 days. Statistical data showed no relationship between survival and the treatment regimen given (p=0.187). The most common side effect of TKI is skin rash. This research should be carried out with a larger sample to minimize bias.

Abstract 5477: An investigation to study differential gene expression of lung cancer cells, tumor associated macrophages, and bone marrow derived mesenchymal stems cells in the tumor microenvironment

Abstract Cancer is a public health crisis affecting approximately 19.3 million globally, annually. Lung cancer has the highest percentage of incidence and is responsible for 27% of cancer-related deaths. The prevalence of lung cancer worldwide led us to investigate differential gene expression in lung adenocarcinoma and small cell lung cancer cell lines in comparison to a healthy lung cell line. We also investigated the effect of lung tumor microenvironment to bone marrow derived pluripotent mesenchymal stem cells (PMSC) and tumor associated macrophages. This research involved cancer and healthy cell line culture with appropriate medium. Whole transcriptomic and small non-coding RNA sequencing and analysis by Ingenuity Pathway Analysis (Qiagen, USA), Quantitative RT-PCR and ELISA assays. We further studied morphological changes and differential gene expression in extracellular vesicles(exosomes) isolated from the TAM and TME exposed PMCs by atomic force microscopy and RNA sequencing techniques. The results of our investigation showed differential gene expression and novel core canonical pathways in the lung cancer cells, TME exposed TAM, and PMSC. We also observed morphological changes in TME derived exosomes from TAM and PMSC. RNA sequencing results showed differential gene expression of both coding and non-coding RNAs including tRNA fragments. Thus, our findings of this investigation led to identification of multiple cancer related genes and small non-coding microRNA, long non-coding RNA, and tRNA fragments which will have potential diagnostic and therapeutic target value that demands further future research. Acknowledgement: This research is supported by a grant from US NIH-NIGMS- 5T34GM100831-08 and US-NIH Grant# U 54 MD007597-31-5959. Citation Format: Hirendra N. Banerjee, Joseph Hedley, Cheslsey Aurelus, Narendra Banerjee, Kuldeep Rawat, Quentin Reaves, Shennel Brown, Darla Gilmartin, Makieyah Liverman, Jazmine Cuffee, Elizabeth Cagle, Erik Armstrong, Brent Lake, Santosh Mandal, Somiranjan Ghosh, Stephen Beebe, Kamal Asadipour, Christopher Krauss, Santanu Bhattacharya, Tanmay Kulkarni, Zahidur Abedin. An investigation to study differential gene expression of lung cancer cells, tumor associated macrophages, and bone marrow derived mesenchymal stems cells in the tumor microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5477.

Abstract 3583: Novel MRI contrast system using halloysite nanotubes embedded with iron-platinum nanoparticles for the regulation of the CHST11 expression for cancer treatment

Abstract Topic: TB Tumor Biology (TB03) In Vivo Imaging. Multifunctional nanocomposites have been a popular research target in recent years. This research aims to produce magnetic nanocomposites with high saturation magnetization (Ms) and good biocompatibility to improve the disadvantage of iron-oxide was easy decomposition in the biological environment and overcome the unclear contrast problem of T2-weighted magnetic resonance imaging (MRI) of lung adenocarcinoma. In order to fit the suitable size of iron-based nanoparticles embedded into the halloysite nanotubes (HNT), the iron-platinum (FePt) nanoparticles were chosen as the MRI’s core materials. The FePt@HNT nanocomposites have low toxicity, and superparamagnetism is expected to be used in biomedical therapy and magnetic separation for disease treatment and environmental protection. Lung adenocarcinoma cancer cells grow in situ, usually invade, and are frequently recurrent after treatment. Since CHST11-mediated gene regulation may serve as a possible means of reducing tyrosine kinase inhibitor (TKI)-induced mitotic cell death, CHST11 modulation of integrin signaling may help lung cancer cells enhance their resistance to gefitinib. To enable the FePt@HNT nanocomposite to be effective in treating lung tumors, we designed miRNAs that inhibit CHST11 expression in an attempt to diagnose lung adenocarcinoma concurrently for treatment. Keywords: Magnetic resonance imaging, halloysite nanotubes, FePt nanoparticles, CHST11, Lung adenocarcinoma

Abstract 2472: Interdisciplinary method to elucidate the interaction between brain microenvironment and cancer cells

Abstract Cancer cells may escape from host immunity to form distant metastases. We have made an original imaging model of brain metastasis (IBrM) to show a heterogeneous interaction between microglia, the tissue macrophages in the brain, and cancer cells in the brain niche. Here we report that enhanced microglial elimination of cancer cells is a new potential treatment for metastatic brain tumors by inhibiting their engraftment. First, to explore factors that favor survival of cancer cells in the brain microenvironment, mouse lung cancer cells (CMT167) were injected through the internal carotid artery, and after 4, 7, 10, and 13 days, surviving cancer cells in the mouse brain were collected and RNAseq was performed. In particular, we focused on CD24/CD7/PD-L1, a known factor that escapes phagocytosis by macrophages. The expression of CD24/CD47 in cancer cells gradually increased, while that of PD-L1 decreased. Next, we injected adenocarcinoma lung cancer cells (LUCs, CMT167) or breast cancer cells (BCCs, E0711) expressing mCherry via the internal carotid artery of the CX3CR1-EGFP mice, whose microglia had been specifically labeled with EGFP. The microglia and LUCs/BCCs were visualized in vivo simultaneously by two-photon microscopy for 14 days. The tumor fate and microglial response against cancer cells were evaluated at single cell resolution in the IBrM. Microglial phagocytosis was significantly increased by single deletion of CD47 or CD24 compared with WT cells. Combined deletion of CD47 and CD24 also synergically increased microglial phagocytosis, resulting in reduced IBrM and prolonged mouse survival. By contrast, deletion of PD-L1 did not enhance microglial phagocytosis or prolong survival. The pharmacological (PLX3397) or genetic conditional (Siglechdtr het/CX3CR1GFP het) conditional depletion of microglia canceled both the increased phagocytosis and the extended survival time, suggesting that the effect of “don't eat me” signals was microglia dependent. The extended survival time was also observed in IBrM using BALB-c nu/nu, that suggest the effect is not dependent on matured T-lymphocytes.These results indicate that novel function of microglia against cancer cells, and suppression of both CD24 and CD47 in cancer cells and enhances microglial phagocytic activity and suppresses the metastatic formation. We are further developing a method for isolating LUCs fluorescently labeled in vivo using holographic 2-photon microscopy for evaluating single cell gene expression profiles. Using the novel techniques, the spatio-temporal information can be integrated across the molecular, cellular and tissue levels that allow us to further investigate the dynamic behavior of not only tumors, but also microglia and neurons. Citation Format: Takahiro Tsuji, Hiroaki Wake, Mariko Shindo, Rahadian Y. Hartantio, Misuzu Horikoshi, Daisuke Kato, Hiroaki Ozasa, Toyohiro Hirai. Interdisciplinary method to elucidate the interaction between brain microenvironment and cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2472.

ALDOC- and ENO2- driven glucose metabolism sustains 3D tumor spheroids growth regardless of nutrient environmental conditions: a multi-omics analysis

BackgroundMetastases are the major cause of cancer-related morbidity and mortality. By the time cancer cells detach from their primary site to eventually spread to distant sites, they need to acquire the ability to survive in non-adherent conditions and to proliferate within a new microenvironment in spite of stressing conditions that may severely constrain the metastatic process. In this study, we gained insight into the molecular mechanisms allowing cancer cells to survive and proliferate in an anchorage-independent manner, regardless of both tumor-intrinsic variables and nutrient culture conditions.Methods3D spheroids derived from lung adenocarcinoma (LUAD) and breast cancer cells were cultured in either nutrient-rich or -restricted culture conditions. A multi-omics approach, including transcriptomics, proteomics, and metabolomics, was used to explore the molecular changes underlying the transition from 2 to 3D cultures. Small interfering RNA-mediated loss of function assays were used to validate the role of the identified differentially expressed genes and proteins in H460 and HCC827 LUAD as well as in MCF7 and T47D breast cancer cell lines.ResultsWe found that the transition from 2 to 3D cultures of H460 and MCF7 cells is associated with significant changes in the expression of genes and proteins involved in metabolic reprogramming. In particular, we observed that 3D tumor spheroid growth implies the overexpression of ALDOC and ENO2 glycolytic enzymes concomitant with the enhanced consumption of glucose and fructose and the enhanced production of lactate. Transfection with siRNA against both ALDOC and ENO2 determined a significant reduction in lactate production, viability and size of 3D tumor spheroids produced by H460, HCC827, MCF7, and T47D cell lines.ConclusionsOur results show that anchorage-independent survival and growth of cancer cells are supported by changes in genes and proteins that drive glucose metabolism towards an enhanced lactate production. Notably, this finding is valid for all lung and breast cancer cell lines we have analyzed in different nutrient environmental conditions. broader Validation of this mechanism in other cancer cells of different origin will be necessary to broaden the role of ALDOC and ENO2 to other tumor types. Future in vivo studies will be necessary to assess the role of ALDOC and ENO2 in cancer metastasis.

LncRNA ADAMTS9-AS1 inhibits the stemness of lung adenocarcinoma cells by regulating miR-5009-3p/NPNT axis

We sought to extend our observation of LncRNA ADAMTS9-AS1 and to specifically uncover its role on the stemness of lung adenocarcinoma (LUAD) cancer cells. ADAMTS9-AS1 was poorly expressed in LUAD. The high ADAMTS9-AS1 expression was positively associated with overall survival. ADAMTS9-AS1 overexpression attenuated the colony-forming capacity and reduced stem cell-like population of LUAD cancer stem cells (CSCs). Furthermore, ADAMTS9-AS1 overexpression increased E-cadherin expression in addition to the downregulated expressions of Fibronectin and Vimentin in LUAD spheres. In vitro results also confirmed the ADAMTS9-AS1's inhibitory effect on the growth of LUAD cells. Moreover, the antagonistic repression of miR-5009-3p levels with the expression of ADAMTS9-AS1 and NPNT was confirmed. Finally, ADAMTS9-AS1 overexpression curbed the increasing stemness of LUDA-CSC caused by NPNT silencing, thus leading to the suppression of LUAD progression in vitro. Conclusively, ADAMTS9-AS1 negatively controls the LUAD cancer cell stemness progression through regulating miR-5009-3p/NPNT axis.

Multiscale reconstruction of bronchus and cancer cells in human lung adenocarcinoma

BackgroundWhile previous studies primarily focused on the structure of the normal whole mouse lung, the whole bronchus and cytoarchitectural details of the mouse intact lung lobe have been discovered at single-cell resolution. Revealing the sophisticated lung adenocarcinoma structure at three-dimensional (3D) and single-cell level remains a fundamental and critical challenge for the pathological mechanism research of lung adenocarcinoma (LA).MethodsFluorescence micro-optical Sectioning Tomography (fMOST) combined with PI staining were used to obtain the 3D imaging of the human LA tissue at single-cell resolution.ResultsWith a spatial resolution of 0.32 × 0.32 × 1.0 μm3, the dataset of human LA with single-cell precision consists of two channels, each of which contains information about the bronchi and the cytoarchitecture. The bronchial wall is thicker and the lumen is smaller in the cancer tissue, in which its original normal structure is vanished. More solid components, more clustered cancer cells with larger nucleoli, and more significant atypia are found in cancer tissue. In paracancerous tissue, the bronchial wall cells have a monolayer or bilayer structure, cluster along the wall, and are relatively dispersed. Few fibrous structures and occasional dissemination of spread through air spaces (STAS) are observed.ConclusionsBased on the human LA tissue dataset obtained by fMOST and PI staining, the bronchi and cells were reconstructed and visualized. This work provides a technical roadmap for studying the bronchus and cytoarchitectural structure and their spatial relationship in LA tissue, which may help with the understanding of the main histological structure of LA among pathologists.

In Vitro Anticancer Activity of Novel Ciprofloxacin Mannich Base in Lung Adenocarcinoma and High-Grade Serous Ovarian Cancer Cell Lines via Attenuating MAPK Signaling Pathway.

Novel drugs are desperately needed in order to combat a significant challenge due to chemo-therapeutic resistance and bad prognosis. This research aimed to assess the anticancer activity of a newly synthesized ciprofloxacin Mannich base (CMB) on ovarian cancer (OVCAR-3) and lung cancer (A-549) cell lines and to investigate probable involved molecular mechanisms. The cytotoxic and pro-apoptotic impact of CMB on both cell lines was investigated using MTT assay, Annexin V assay, and cell cycle analysis, as well as caspase-3 activation. Western blotting was carried out to evaluate downstream targets of the MAPK pathway, while qRT PCR was used to evaluate the gene expression pattern of the p53/Bax/Bcl2 pathway. CMB treatment showed significantly reduced cell proliferation in both OVCAR-3 and A-549 cells with half maximum inhibitory concentration (IC50) = 11.60 and 16.22 µg/mL, respectively. CMB also induced apoptosis, S phase cell cycle arrest, and up-regulated expression of p53, p21, and Bax while down-regulated Bcl2 expression. CMB also halted cell proliferation by deactivating the MAPK pathway. In conclusion, CMB may be regarded as a potential antiproliferative agent for lung and ovarian cancers due to anti-proliferative and pro-apoptotic actions via inhibition of the MAPK pathway and p53/Bax/Bcl2.

Anticancer property of marine coral-derived Streptomyces sp. SS162 against A549 lung adenocarcinoma cancer cells

Anticancer property of marine coral-derived Streptomyces sp. SS162 against A549 lung adenocarcinoma cancer cells

Characterization and in vitro anticancer potential of exopolysaccharide extracted from a freshwater diatom Nitzschia palea (Kütz.) W.Sm. 1856

Diatoms are photoautotrophic microalgae classified under class Bacillariophyceae, engulfed by hard silicate frustules, which give mechanical support and protection from bacterial infections. They exude polysaccharides extracellularly that help them with their gliding motion (locomotion). However, the bioactivity of such compounds was least explored from freshwater diatoms. In the present study, a single species of pennate diatom identified as Nitzschia palea was isolated and molecularly characterized by 18S rRNA smaller subunit gene (partial) sequencing and submitted to GenBank NCBI and accession number retrieved as ON360983. Based on logarithmic growth curve analysis, the exponential phase was obtained from 3rd to 4th day of diatom culture. The exopolysaccharide was extracted by the hot-water extraction method, and characterized by FT-IR. The total yield of exopolysaccharide from Nitzschia palea was estimated as 1.56 mg in 100 mL of culture after 7 days of incubation. The estimated carbohydrate content was 51.35 µg/100 µL. The monosaccharide constituents were determined by acid hydrolysis of exopolysaccharide, silylation (derivatization), followed by GC–MS analysis and tabulated. The extracted exopolysaccharide was evaluated for its anti-cancer potential against the Human Adenocarcinoma lung cancer cell line (A549) and the estimated IC50 value was 62.64 µg/mL. Acridine orange staining assay and DNA fragmentation assay also confirmed the apoptotic activity of exopolysaccharide derived from the diatom Nitzschia palea.

Exploration of 1,2,3-triazole linked benzenesulfonamide derivatives as isoform selective inhibitors of human carbonic anhydrase

A novel series of 1,2,3-triazole benzenesulfonamide substituted 1,3-dioxoisoindolin-5-carboxylate (7a-l) inhibitors of human α-carbonic anhydrase (hCA) was designed using a tail approach. The design method relies on the hybridization of a benzenesulfonamide moiety with a tail of 1,3-dioxoisoindoline-5-carboxylate and a zinc-binding group on a 1,2,3-triazole scaffold. Among the synthesized analogues, 2‑iodophenyl (7f, KI of 105.00 nM and SI of 2.98) and 2‑naphthyl (7h, KI of 32.11 nM and SI of 3.48) analogues (over off-target hCA I) and phenyl (7a, KI of 50.13 nM and SI of 2.74) and 2,6‑dimethylphenyl (7d, KI of 50.60 nM and SI of 3.35) analogues (over off-target hCA II) exhibited a remarkable selectivity for tumor isoforms hCA IX and XII, respectively. Meanwhile, analogue 7a displayed a potent inhibitory effect against the tumor-associated isoform hCA IX (KI of 18.29 nM) compared with the reference drug acetazolamide (AAZ, KI of 437.20 nM), and analogue 7h showed higher potency (KI of 9.22 nM) than AAZ (KI of 338.90 nM) against another tumor-associated isoform hCA XII. However, adding the lipophilic large naphthyl tail to the 1,3-dioxoisoindolin-5-carboxylate analogues increased both the hCA inhibitory and selective activities against the target isoform, hCA XII. Additionally, these analogues (7a-l) showed IC50 values against the human lung (A549) adenocarcinoma cancer cell line ranging from 129.71 to 352.26 μM. The results of the molecular docking study suggested that the sulfonamide moiety fits snugly into the hCAs active sites and interacts with the Zn2+ ion. At the same time, the tail extension engages in various hydrophilic and hydrophobic interactions with the nearby amino acids, which affects the potency and selectivity of the hybrids.

In Vitro Antiproliferative Activity of Ptaeroxylon obliquum Leaf Extracts, Fractions and Isolated Compounds on Several Cancer Cell Lines

Several cancers are induced by microbial infections or chronic inflammation. Ptaeroxylon obliquum is traditionally used to treat various infections characterized by inflammation. The in vitro antiproliferative and antioxidant activity of P. obliquum leaf extracts, fractions and isolated compounds were determined. Antiproliferative activity was assessed against normal Vero cells, and several cancerous human cells, including human breast cancer (MCF-7), hepatocarcinoma (HepG2), lung adenocarcinoma (A549) and human cervical cancer cells (HeLa) using a colorimetric tetrazolium bromide assay. Radical scavenging activity was tested using the 2,2-diphenyl-1-instrpicrylhydrazyl (DPPH) and 2,2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. Obliquumol, O-methylalloptaeroxylin and a mixture of lupeol and β-amyrin were isolated from the chloroform fraction using silica gel open column chromatography. Acetone extracts were toxic to HepG2 cells with IC50 values from 8 to 200 µg/mL but were less toxic to other cells with selectivity index as high as 14. Aqueous extracts and fractions were non-toxic at concentrations tested against all the cell lines (IC50 > 100 µg/mL). Isolated compounds had IC50 values ranging from 52 to 539 µg/mL and 189 to 247 µg/mL against HepG2 and HeLa cells, respectively. Light microscopy showing changes in HepG2 and HeLa cell morphology supported the cytotoxicity of the acetone extracts. Water extracts scavenged ABTS and DPPH radicals with IC50 values as low as 29.06 µg/mL and 43.4 µg/mL. P. obliquum extracts may be useful as sources of anticancer therapy, as they have selective cytotoxicity against cancer cell lines.