Immunohistochemical Analysis of Human Lung Cancers with Anti-ras p21 Monoclonal Antibodies

The expression of ras oncogene product p21 in human malignant pleurisy and primary lung cancer was studied immunocyto-histochemically with monoclonal antibodies (MoAbs) rp-28 and rp-35 against ras p21. In pleural effusion cells, cancer cells revealed more intensively positive reaction with MoAb rp-35 than with MoAb rp-28, especially in the plasma membrane, and no positive reaction was obtained in any kind of inflammation cells with the exception of faintly positive reaction in the cytoplasm of macrophages. In primary lung cancers, well or moderately differentiated adenocarcinoma tissues showed higher reactivity with MoAb rp-28 than those of poorly differentiated adenocarcinoma or any other histological subtype of lung cancer. With MoAb rp-35, intensively positive reaction was obtained in most of cases with all different histological subtypes of lung cancer. The staining in cancer cells was usually localized intensively to the plasma membrane and weakly to the cytoplasm with both MoAbs. Normal bronchial epithelial and glandular tissues showed only cytoplasmic staining. These two MoAbs, especially MoAb rp-35, may be useful in clinicopathological applications for the diagnosis of malignant pleurisy and primary lung cancer.

Download Full-text

Tubulin acetylation enhances lung cancer resistance to paclitaxel-induced cell death through Mcl-1 stabilization

Cell Death Discovery ◽

10.1038/s41420-021-00453-9 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Onsurang Wattanathamsan ◽

Rawikorn Thararattanobon ◽

Ratchanee Rodsiri ◽

Pithi Chanvorachote ◽

Chanida Vinayanuwattikun ◽

...

Keyword(s):

Lung Cancer ◽

Cell Death ◽

Cancer Cells ◽

Trichostatin A ◽

Primary Lung Cancer ◽

Lung Cancer Cells ◽

Cancer Resistance ◽

Apoptosis Resistance ◽

Tubulin Acetylation ◽

Cancer Aggressiveness

AbstractThe posttranslational modifications (PTMs) of microtubules have been reported to play an important role in cancer aggressiveness, including apoptosis resistance. In this study, we aimed to investigate the biological role of microtubule PTMs in the regulation of paclitaxel responsiveness. The acetylated tubulin (Ace-tub) level was strongly associated with paclitaxel sensitivity, as observed in patient-derived primary lung cancer cells and xenografted immunodeficient mice. We showed that paclitaxel-resistant H460 lung cancer cells, generated by a stepwise increase in paclitaxel, exhibited markedly increased tubulin acetylation and consequently acquired paclitaxel resistance. Upregulation of tubulin acetylation by overexpression of α-tubulin acetyltransferase 1 wild-type (αTAT1wt), an enzyme required for acetylation, or by treatment with trichostatin A (TSA), a histone deacetylase 6 (HDAC6) inhibitor, significantly attenuated paclitaxel-induced apoptosis. Investigation of the underlying mechanism revealed that the levels of antiapoptotic Mcl-1 appeared to increase in αTAT1wt-overexpressing and TSA-treated cells compared to control cells, whereas the levels of other antiapoptotic regulatory proteins were unchanged. On the other hand, decreased tubulin acetylation by αTAT1 RNA interference downregulated Mcl-1 expression in patient-derived primary lung cancer and paclitaxel-resistant lung cancer cells. A microtubule sedimentation assay demonstrated that Mcl-1 binds to microtubules preferentially at Ace-type, which prolongs the Mcl-1 half-life (T1/2). Furthermore, immunoprecipitation analysis revealed that polyubiquitination of Mcl-1 was extensively decreased in response to TSA treatment. These data indicate that tubulin acetylation enhances the resistance to paclitaxel-induced cell death by stabilizing Mcl-1 and protecting it from ubiquitin–proteasome-mediated degradation.

Download Full-text

Quiescin Sulfhydryl Oxidase 1 (QSOX1) Secreted by Lung Cancer Cells Promotes Cancer Metastasis

International Journal of Molecular Sciences ◽

10.3390/ijms19103213 ◽

2018 ◽

Vol 19 (10) ◽

pp. 3213 ◽

Cited By ~ 5

Author(s):

Hye-Jin Sung ◽

Jung-Mo Ahn ◽

Yeon-Hee Yoon ◽

Sang-Su Na ◽

Young-Jin Choi ◽

...

Keyword(s):

Lung Cancer ◽

Cancer Cells ◽

Cancer Metastasis ◽

Lung Cancer Cells ◽

Migration And Invasion ◽

Sulfhydryl Oxidase ◽

Lung Cancers ◽

Normal Tissues ◽

Cancer Tissues ◽

Quiescin Sulfhydryl Oxidase

As lung cancer shows the highest mortality in cancer-related death, serum biomarkers are demanded for lung cancer diagnosis and its treatment. To discover lung cancer protein biomarkers, secreted proteins from primary cultured lung cancer and adjacent normal tissues from patients were subjected to LC/MS–MS proteomic analysis. Quiescin sulfhydryl oxidase (QSOX1) was selected as a biomarker candidate from the enriched proteins in the secretion of lung cancer cells. QSOX1 levels were higher in 82% (51 of 62 tissues) of lung cancer tissues compared to adjacent normal tissues. Importantly, QSOX1 serum levels were significantly higher in cancer patients (p < 0.05, Area Under curve (AUC) = 0.89) when measured by multiple reaction monitoring (MRM). Higher levels of QSOX1 were also uniquely detected in lung cancer tissues, among several other solid cancers, by immunohistochemistry. QSOX1-knock-downed Lewis lung cancer (LLC) cells were less viable from oxidative stress and reduced migration and invasion. In addition, LLC mouse models with QSOX1 knock-down also proved that QSOX1 functions in promoting cancer metastasis. In conclusion, QSOX1 might be a lung cancer tissue-derived biomarker and be involved in the promotion of lung cancers, and thus can be a therapeutic target for lung cancers.

Download Full-text

Escape from Therapy-Induced Accelerated Cellular Senescence in p53-Null Lung Cancer Cells and in Human Lung Cancers

Cancer Research ◽

10.1158/0008-5472.can-04-1270 ◽

2005 ◽

Vol 65 (7) ◽

pp. 2795-2803 ◽

Cited By ~ 199

Author(s):

Rachel S. Roberson ◽

Steven J. Kussick ◽

Eric Vallieres ◽

Szu-Yu J. Chen ◽

Daniel Y. Wu

Keyword(s):

Lung Cancer ◽

Cancer Cells ◽

Cellular Senescence ◽

Human Lung ◽

Lung Cancer Cells ◽

Lung Cancers

Download Full-text

Emetine Synergizes with Cisplatin to Enhance Anti-Cancer Efficacy against Lung Cancer Cells

International Journal of Molecular Sciences ◽

10.3390/ijms20235914 ◽

2019 ◽

Vol 20 (23) ◽

pp. 5914 ◽

Cited By ~ 1

Author(s):

Ti-Hui Wu ◽

Shan-Yueh Chang ◽

Yu-Lueng Shih ◽

Tsai-Wang Huang ◽

Hung Chang ◽

...

Keyword(s):

Lung Cancer ◽

Cancer Cells ◽

Cisplatin Resistance ◽

Clinical Problem ◽

Driver Mutations ◽

Anticancer Efficacy ◽

Lung Cancers ◽

Anticancer Effects ◽

Therapeutic Choice ◽

Reporter Assays

Cisplatin is still the primary therapeutic choice for advanced lung cancers without driver mutations. The occurrence of cisplatin resistance is a major clinical problem in lung cancer treatment. The natural extracted agent emetine reportedly has anticancer effects. This study aimed to explore the possible role of emetine in cisplatin resistance. We used cell viability, Western blot, and Wnt reporter assays to show that emetine suppresses proliferation, β-catenin expression, and Wnt/β-catenin signaling in non-small cell lung cancer (NSCLC). The synergism of emetine and cisplatin was assessed by constructing isobolograms and calculating combination index (CI) values using the Chou-Talalay method. Emetine effectively synergized with cisplatin to suppress the proliferation of cancer cells. Furthermore, nuclear β-catenin and cancer stem cell-related markers were upregulated in the cisplatin-resistant subpopulation of CL1-0 cells. Emetine enhanced the anticancer efficacy of cisplatin and synergized with cisplatin in the cisplatin-resistant subpopulation of CL1-0 cells. Taken together, these data suggest that emetine could suppress the growth of NSCLC cells through the Wnt/β-catenin pathway and contribute to a synergistic effect in combination with cisplatin.

Download Full-text

Comprehensive cancer genomics-based screening of new therapeutic targets and biomarkers for lung cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.e21031 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. e21031-e21031

Author(s):

Yataro Daigo ◽

Atsushi Takano ◽

Yusuke Nakamura

Keyword(s):

Lung Cancer ◽

Cancer Cells ◽

Cancer Patients ◽

Therapeutic Target ◽

Cancer Genomics ◽

Cancer Biomarkers ◽

Lung Cancer Cells ◽

Lung Cancers ◽

Lung Cancer Patients ◽

Target Molecules

e21031 Background: Since the clinical outcome of advanced lung cancer patients is still poor after standard therapies, development of new anti-cancer drugs with minimum risk of adverse effects and cancer biomarkers for precision medicine is urgently required. Methods: We have been screening new therapeutic target molecules and molecular biomarkers for lung cancers as follows; i) To identify overexpressed genes in lung cancers by the gene expression profile analysis, ii) To verify the target genes for their scarce expression in normal tissues, iii) To validate the clinicopathologic importance of their protein expression by tissue microarray covering 263 lung cancers, and iv) To confirm their function for the growth and/or invasive ability of the lung cancer cells by siRNAs and gene transfection assays. Results: We identified dozens of candidate target molecules and selected a gene encoding protein with a GAP domain, LAPG1 (lung cancer-associated protein with Gap domain 1). Immunohistochemical analysis showed that LAPG1 expression was observed in 69.9% of lung cancers. Moreover positivity of LAPG1 expression was associated with poor prognosis of lung cancer patients. Knockdown of LAPG1 expression by siRNAs suppressed growth of lung cancer cells. Introduction of LAPG1 increased the invasive activity of mammalian cells, indicating that LAPG1 could be a prognostic biomarker and therapeutic target for lung cancers. Conclusions: Comprehensive cancer genomics-based screening could be useful for selection of new cancer biomarkers and molecular targets for developing small molecules, antibodies, nucleic acid drugs, and immunotherapies.

Download Full-text

Does multiplicity of synchronous multiple lung primaries (SMLPs) affect prognosis.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.e20530 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. e20530-e20530

Author(s):

RuoBing Xue ◽

Satish Maharaj ◽

Rohit Kumar ◽

Goetz H. Kloecker

Keyword(s):

Lung Cancer ◽

Adjuvant Therapy ◽

Early Stage ◽

Primary Lung Cancer ◽

Case Series ◽

Current Recommendation ◽

Lung Cancers ◽

T Stage ◽

Wide Range ◽

Common Risk Factors

e20530 Background: SMLPs are detected more commonly due to advancements in screening technology. Their workup and classification; however, are still lacking a clear standard. T stage of the largest lesion has been used as the major prognostic marker. This; however, does not take the number of SMLPs and their genomic drivers into consideration. This study aims to identify and review common risk factors associated with SMLPs and determine whether the number of primaries influence the prognosis. Methods: A systematic review of the literature published between 2000 and 2021 was conducted through PubMed and Medline by using the combination of keywords, including: “synchronous multiple primary lung cancer”, “simultaneous multifocal lung cancer”, “synchronous solitary lung metastasis”, “risk factor” and “prognosis”. A total of fifty studies were identified, among them only sixteen retrospective research articles and two review articles were relevant to the study at hand. Results: Sixteen retrospective studies including a total of 1685 eligible patients were reviewed. Thirteen of these studies reported the main histology type to be adenocarcinoma with a ratio ranging from 35% to 96.8%. Eight studies have reported the numbers of synchronous primary lung cancers, including one study found 11 SMLPs. Among these, one study by van Rens found number of SMLPs impact prognosis adversely compared to a single lung cancer. However, three other studies demonstrated multiple SMPLs do not adversely affect survival (Finley et al, 2010; Kocaturk et al, 2011; Li et al, 2020). Four of the sixteen studies analysed the effect of multiple lobes involvement and distance between tumors, with varying conclusions; two studies reported no difference in prognosis while one study revealed worse survival with multiple lobe involvement and one study found favorable outcome. Most studies confirm the usual prognostic factors for SMLPs, including: gender, smoking, type of surgery, comorbidities and adjuvant therapy. The median 5 year OS reported for SMLPs is 66%, with a wide range from 19% to 95.8%.The 3 year OS is 75% in most studies. Conclusions: The data on how the number of SMLPs affects the prognosis is uncertain. The current recommendation to base the decision for adjuvant therapy on the highest T stage is not supported by prospective evidence or consistent among published case series. Considering the recent approval of targeted therapies in early stage lung cancers, a better prognostic scoring system for SMLPs is required.

Download Full-text

The rates of second lung cancers and the survival of surgically-resected second primary lung cancers in patients undergoing resection of an initial primary lung cancer

Lung Cancer ◽

10.1016/j.lungcan.2020.07.015 ◽

2020 ◽

Vol 147 ◽

pp. 115-122

Author(s):

J.M. Varlotto ◽

R. Voland ◽

M.M. DeCamp ◽

Paul Rava ◽

T.J Fitzgerald ◽

...

Keyword(s):

Lung Cancer ◽

Primary Lung Cancer ◽

Second Primary ◽

Lung Cancers

Download Full-text

Genomics and proteomics-based discovery of novel cancer biomarkers and molecular targets for cell-permeable peptide/small molecule-based drugs.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.15_suppl.e13552 ◽

2012 ◽

Vol 30 (15_suppl) ◽

pp. e13552-e13552

Author(s):

Yataro Daigo ◽

Atsushi Takano ◽

Yusuke Nakamura

Keyword(s):

Lung Cancer ◽

Cancer Cells ◽

Tissue Microarray ◽

Small Molecule ◽

Phosphorylation Site ◽

Lung Cancer Cells ◽

Lung Cancers ◽

Genomics And Proteomics ◽

Cell Permeable Peptide ◽

Erk Kinase

e13552 Background: Identification of cancer-specific oncoproteins is an effective approach to develop new diagnostics and therapeutics. Methods: We have established a strategy as follows to identify new oncoproteins, which can be applied as biomarkers and drug development; i) To identify genes overexpressed in 120 clinical lung cancers using the cDNA microarray representing 27,648 genes, ii) To verify the genes for their low expression in 23 normal tissues by northern-blotting, iii) To validate the clinicopathological significance of their protein expression by tissue microarray covering 262 cases of non-small cell lung cancers (NSCLCs), iv) To verify whether they are essential for the growth of cancer cells by siRNAs, v) To do immunoprecipitation assays and mass-spectrometric analysis to identify their interacting proteins in cancer cells, and screening of cell-permeable peptides that could inhibit the protein-protein interaction that is essential for carcinogenesis. Results: We identified 35 oncoproteins to be upregulated in the majority of lung cancers, and further selected CDCA5 (cell division cycle associated 5) as a good candidate. Tumor tissue microarray analysis of 262 NSCLC patients revealed that CDCA5 positivity was an independent prognostic factor. Suppression of CDCA5 expression with siRNAs inhibited the growth of lung cancer cells; concordantly, induction of exogenous expression of CDCA5 conferred growth-promoting activity in mammalian cells. We also found that extracellular signal-regulated kinase (ERK) kinase interacted with and phosphorylated CDCA5 at Serine 209 in vivo. Functional inhibition of the interaction between CDCA5 and ERK kinase by a cell-permeable peptide corresponding to a 20-amino-acid sequence part of CDCA5, which included the Serine 209 phosphorylation site by ERK, significantly reduced phosphorylation of CDCA5 and resulted in growth suppression of lung cancer cells. Conclusions: CDCA5 positivity should be useful as a novel prognostic biomarker in the clinic. Selective suppression of the ERK-CDCA5 pathway by cell-permeable peptide or small molecule-based drugs could be a promising strategy for cancer therapy.

Download Full-text

Systematic approach for development of new immunotherapeutics for lung cancers through cancer genomics analysis.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.3092 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. 3092-3092

Author(s):

Yataro Daigo ◽

Atsushi Takano ◽

Yusuke Nakamura

Keyword(s):

Lung Cancer ◽

Phase I ◽

Cancer Cells ◽

Cancer Patients ◽

Clinical Studies ◽

Critical Role ◽

Phase I Clinical Trials ◽

Lung Cancers ◽

Lung Cancer Patients ◽

High Immunogenicity

3092 Background: Oncoantigens are defined to be proteins that are very specifically expressed in cancer cells and that have the oncogenic activity and high immunogenicity, and are considered to be promising targets for immunotherapy such as therapeutic cancer vaccines. Methods: We have established a strategy as follows to identify new oncoantigens; i) screening of highly transactivated genes in the majority of 120 lung cancers using cDNA microarray representing 27,648 genes coupled with enrichment of tumor cells by laser microdissection, ii) verification of no expression of each candidate gene in normal tissues by northern-blot analysis, iii) validation of the clinicopathological significance of its high level of expression with tissue microarray containing 300 lung cancers, iv) verification of a critical role of each gene in the growth or invasiveness of cancer cells by RNAi and cell growth/invasion assays, v) screening of the epitope peptides recognized by HLA-A*0201- or A*2402-restricted cytotoxic T lymphocyte (CTL). We conducted phase I clinical trials of these therapeutic peptide vaccines for lung cancer patients. Results: We identified 35 oncoantigens and screened dozens of 10-amino-acid peptides, each of which corresponded to a part of TTK, LY6K, IMP-3, CDCA1, KIF20A, CDC45L, and FOXM1, and was a candidate to be presented on the surface of HLA-A*0201 or HLA-A*2402 that induced in vitro CTL response. Phase I clinical studies indicated that five epitope peptides could strongly induce the CTL activity in cancer patients. For example, we conducted a phase I study for HLA-A*2402-positive, advanced non-small cell lung cancer patients who failed to standard therapy, using the combination of 1, 2 or 3 mg/body of each peptides from LY6K, CDCA1, and KIF20A mixed with adjuvant once a week. This cancer vaccine therapy demonstrated tolerability and had very high immunogenicity of even 1 mg/body dose to induce antigen-specific CTLs in cancer patients. Conclusions: Through systematic genomics-based approach and clinical study, we have identified five epitope peptides, which could induce CTLs very effectively in cancer patients, and therefore it warrants further clinical studies. Clinical trial information: NCT01069575.

Download Full-text