Proteins In Squamous Cell Carcinoma Research Articles

Novel Schiff Bases: Synthesis, Characterization, Bioactivity, Cytotoxicity, and Computational Evaluations

Hypopharyngeal cancer is rare subtype of head and neck cancers with relatively poor prognosis. Current therapeutic modalities lack the potential to provide patients with better clinical outcome and quality of life. This study was conducted on the synthesis of 2-methoxy-4-((3-alkyl(aryl)-5-oxo-1H-1,2,4-triazol-4(5H)-ylimino)-methyl)-phenyl-4-(2-methoxy-4-((3-alkyl(aryl)-5-oxo-1H-1,2,4-triazol-4(5H)-ylimino)-methyl)-phenyl)-4-oxobutanoates (3) using biologically important 1,2,4-triazole. The condensation of 3-alkyl(aryl)-4-amino-4,5-dihydro-1H-1,2,4-triazol-5-ones (1) with 4-formyl-2-methoxyphenyl-4-(4-formyl-2-methoxyphenyl)-4-oxobutanoate yielded the biologically active 2-methoxy-4-((3-alkyl(aryl)-5-oxo-1H-1,2,4-triazol-4(5H)-ylimino)-methyl)-phenyl-4-(2-methoxy-4-((3-alkyl(aryl)-5-oxo-1H-1,2,4-triazol-4(5H)-ylimino)-methyl)-phenyl)-4-oxobutanoates (3). The compounds obtained were analyzed via FT-IR,1H-/13C-NMR spectrometers, elemental analysis, and HRMS spectroscopic techniques. Furthermore, we aimed at investigating the potential of compounds 3a-g against FaDu hypopharyngeal cancer cells. We demonstrated that compounds 3c, 3e, and 3 g had relatively lower IC50 values compared to the remaining tested compounds and more importantly their IC50 values were comparable to 5-FU, which suggests them as important therapeutic agent candidates. These newly synthesized compounds were assessed for their inhibitory activities toward two human carbonic anhydrase isoforms I and II (hCA I and II). Then, molecular docking calculations were made to compare the biological activities of studied molecules against cancer proteins. Compound 3c has a docking score of −7.15 against squamous cell carcinoma protein with ID: 2DO4 and −5.49 docking score against squamous cell carcinoma protein with ID:5PJZ. ADME/T analysis was performed to examine the drug properties of studied molecules.

Association of IHC p16INK4a expression and ELISA Plasma p16INK4a Protein in Squamous Cell Carcinoma of Uterine Cervix: A Concept of Liquid Biopsy

Introduction: Cervical cancer is the 3rd most common cancer among women worldwide. p16 biomarker is the surrogate marker in cervical cancer and can be detected by Immunohistochemistry (IHC). Aim: To evaluate the association of IHC p16INK4a expression in tissue sections and Enzyme-linked Immunosorbent Assay (ELISA) p16INK4a protein in plasma of paired samples in Squamous Cell Carcinoma (SCC) of uterine cervix. Materials and Methods: This laboratory-based observational, pilot study was conducted at Sri Devaraj Urs Medical College (tertiary health care centre) in South India from June 2020 to May 2021. Total 17 new cases of SCC of cervix diagnosed by histopathology were considered for the study. The cases were staged as per International Federation of Gynecology and Obstetrics (FIGO) staging system and classified by histopathology as keratinising and non keratinising types. IHC p16INK4a evaluation was done on tissue sections and classified as block positivity, ambiguity and negative. ELISA p16INK4a estimation was done using plasma from Dipotassium Ethylenediaminetetraacetic Acid (K2-EDTA) blood sample of same cases. Statistical analysis was done using Statistical Package for the Social Sciences (SPSS) version 22.0 software. Results: The age ranged from 30-80 years with mean of 54.3±12.0 years. Plasma ELISA p16INK4a level ranged from 3.4-19.6 ng/ mL with a mean of 7.2±2.35 ng/mL in SCC of cervix. The plasma p16INK4a ELISA levels of 5.1 to 6.2 ng/mL, 6.0 to 6.6 ng/mL and 5.5 to 9.7 ng/mL predicts negative, ambiguity and block positivity of IHC p16INK4a expression respectively in corresponding tissue biopsy. Plasma ELISA p16INK4a levels were maximum in Non Keratinising Squamous Cell Carcinoma (NKSCC) followed by Well Differentiated Squamous Cell Carcinoma (WDSCC), Moderately Differentiated Squamous Cell Carcinoma (MDSCC) and Poorly Differentiated Squamous Cell Carcinoma (PDSCC). Conclusion: The plasma ELISA p16INK4a levels and IHC p16INK4a expression were maximum in higher disease stage compared to lower stage. This was a pilot study to evaluate the association between tissue IHC p16 expression and plasma ELISA p16 levels. Further study has to be done on larger study population with standardised procedure to prove the hypothesis.

Evaluation of enzyme-linked immunosorbent assay plasma p16INK4a protein in squamous cell carcinoma in uterine cervix

Cervical cancer is one of the common cancers in females. The common method of screening is Pap test which has low sensitivity. Hence, better methods are explored with different biomarkers, of which estimation of P16 protein can be opted in early detection of cervical cancer. Seventy cases and seventy controls were considered for the study. Cases were invasive squamous cell carcinoma (SCC) of cervix confirmed by histopathology. Controls were healthy age-matched females. The blood sample of cases and controls was collected in K2 Ethylenediaminetetraacetic acid vacutainer, and the separated plasma was subjected to estimation of P16 protein by quantitative sandwich Enzyme-Linked ImmunoSorbent Assay method. The data were analyzed for the association between p16 protein in plasma in cases and controls. The age among cases and controls ranged from 30 to 80 years. The P16 levels among cases ranged from 3.4 to 19.6 ng/ml with a mean of 7.24 ± 2.35 ng/ml. The plasma P16 level in controls ranged between 0.9 and 9.7 ng/ml with mean of 4.1 ± 2.22 ng/ml. At cutoff more than 4.8 ng/ml in cases, the sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy were 92.86%, 72.86%, 77.4%, 91.1%, and 82.86%, respectively. The specificity increased with increase in plasma p16 levels. The P16 levels were maximum in stage IV disease. This was a pilot study to detect the plasma p16INK4a levels in SCC of cervix. The levels of plasma p16 protein between 3.9 and 5 ng/ml can be considered as the range for the test to be positive. In clinically suspected cases of cervical cancer, levels more than 4.8 ng/ml can be considered for the diagnosis as point of care test.

Immunohistochemical Expression of Epithelial Mesenchymal Transition Proteins in Squamous Cell Carcinoma of the Oral Cavity

Background & Objective: Epithelial-Mesenchymal transition (EMT) is known to be a possible mechanism in tumor progression; however, there is insufficient evidence to support the contribution of this process in human cancers. The present study aimed to evaluate the expression of EMT markers in normal oral epithelium and oral squamous cell carcinoma and also correlates with some clinicopathological parameters.Methods: This study was conducted on 70 samples, including 20 cases of normal epithelium and 50 cases of Oral Squamous cell Carcinoma (OSCC). To examine the expression level of these proteins, immunohistochemical staining was performed for samples using E-cadherin and N-cadherin monoclonal antibodies.Results: Reduced expression of E-cadherin was observed in 74% of OSCC and 15% of normal epithelium samples; this difference was statistically significant (P˂0.000). With the progression of SCC from well towards poor differentiation, the E-cadherin expression decreased; however, this difference was not statistically significant (P=0.642). Normal epithelial specimens were negative for N-cadherin expression in 75% of cases, whereas OSCC specimens showed high expression of N-cadherin in 46% of cases, this difference was statistically significant (P=0.01). Although 62.5% of poorly differentiated OSCC showed high expression of N-cadherin, the difference between the histopathological grades was not significant (P=0.586). No significant relationship was found between markers expression and patient’s age, gender, and tumor location.Conclusion: This study showed that OSCC tissues showed high EMT phenotype (reduced E-cadherin expression and high expression of N-cadherin) compared to normal oral mucosa which may indicate the possible key role of EMT mechanism during oral carcinogenesis.

Immunohistochemical expression of bcl-2; an apoptosis regulatory protein in squamous cell carcinoma of oropharynx: A diagnostic cross-sectional study

BackgroundThe Oropharyngeal squamous cell carcinoma is the second most common head and neck malignancy. Bcl-2 expression alterations have been reported invariably in different cancers. It plays a part in carcinogenesis by inhibiting programmed cell death and thus increasing cell survival. Materials and methodsThe present study was conducted in Department of Pathology, S.G.T. Medical College and University, Gurugram over a period of one year (2019–20) on biopsy proven cases of squmaous cell carcinoma of oropharynx. Grading of the tumor was done using Anneroth's multifactorial grading system. The Bcl-2 scoring was done. ResultsIn the present study, a total of 75 cases of oropharyngeal SCC constituted the study group, with the mean age at presentation of 56.63 years. The correlation between Anneroth's grading system and WHO grade was found to be statistically significant, while correlation between WHO grade with lymph node status was found to be statistically non significant. ConclusionThere was significant correlation between Anneroth's grading system and WHO grading system in SCC of oropharynx and it was found to be more relevant in predicting the stage of tumor. Bcl-2 expression did not correlate with the grading of tumor.

Immunohistochemical Expression of Epithelial Mesenchymal Transition Proteins in Squamous Cell Carcinoma of the Oral Cavity.

Background & Objective:Epithelial-Mesenchymal transition (EMT) is known to be a possible mechanism in tumor progression; however, there is insufficient evidence to support the contribution of this process in human cancers. The present study aimed to evaluate the expression of EMT markers in normal oral epithelium and oral squamous cell carcinoma and also correlates with some clinicopathological parameters.Methods:This study was conducted on 70 samples, including 20 cases of normal epithelium and 50 cases of Oral Squamous cell Carcinoma (OSCC). To examine the expression level of these proteins, immunohistochemical staining was performed for samples using E-cadherin and N-cadherin monoclonal antibodies.Results:Reduced expression of E-cadherin was observed in 74% of OSCC and 15% of normal epithelium samples; this difference was statistically significant (P˂0.000). With the progression of SCC from well towards poor differentiation, the E-cadherin expression decreased; however, this difference was not statistically significant (P=0.642). Normal epithelial specimens were negative for N-cadherin expression in 75% of cases, whereas OSCC specimens showed high expression of N-cadherin in 46% of cases, this difference was statistically significant (P=0.01). Although 62.5% of poorly differentiated OSCC showed high expression of N-cadherin, the difference between the histopathological grades was not significant (P=0.586). No significant relationship was found between markers expression and patient’s age, gender, and tumor location. Conclusion:This study showed that OSCC tissues showed high EMT phenotype (reduced E-cadherin expression and high expression of N-cadherin) compared to normal oral mucosa which may indicate the possible key role of EMT mechanism during oral carcinogenesis.

MRNA expression of ferroptosis-related proteins in squamous cell carcinoma of tongue

Objective: Glossal Squamous Cell Carcinoma (GSCC) has a distinct molecular profile including alterations in programed regulated cell death (RCD). Ferroptosis is a newly reported form of RCD mediated by ferric ions and mediated by several proteins. Till date, the possibility of this phenomenon in GSCC has not been explored. Materials and Methods: The proteins associated with ferroptosis were collated. The mRNA of corresponding proteins expression from GSCC lesional tissues and control tissues from the same patient from the human cancer genome project was obtained as raw values and subjected to differential expression (DE) analysis using DESeq2 statistics. The Benjamini and Hochberg (false discovery rate) approach was used to adjust P-value and P ≤ 0.05 was set at statistical significance. Log fold change was set at 0.6. The significant genes were subjected to network analysis for protein-protein interaction and this network was analyzed for pathway enrichment analysis and presented. Results: There were 12 cases that had GSCC and controls (n = 12). There were 40 genes directly involved with ferroptosis and another 50 genes associated with the ferroptosis. Of this, 28 genes had DE, of which 16 genes were directly involved in the ferroptosis pathway and 12 genes associated with it. These genes, in enrichment analysis, indicated that the other pathways involved several other cancers and cancer signaling. Certain pro-ferroptotic genes and anti-ferroptotic genes had DE. Increase of PTGS2 and ACSL4 mRNA expression in GSCC tissue as compared to control was a significant finding. There is an increase of mRNA of genes that increase the ferric ions intracellularly. These findings would help to draw better and effective treatment strategies that can help to target neoplastic cells in GSCC. Conclusion: In GSCC, ferroptosis is a possibility and the differential expression of certain pro-ferroptotic genes could he harnessed to draw effective and safer treatment strategies.

Decrease in phospho-PRAS40 plays a role in the synergy between erlotinib and crizotinib in an EGFR and cMET wild-type squamous non-small cell lung cancer cell line

IntroductionLung squamous cell carcinomas (SCC) typically harbor a strong activation of epidermal growth factor receptor (EGFR) pathway. Since one of the most common resistance mechanisms against EGFR inhibition relies on the activation of cMET parallel signaling, we investigated the efficacy of a dual blockade with erlotinib and crizotinib in EGFR and cMET wild-type lung SCC cell lines. MethodsDrug sensitivity assays were performed on LUDLU, SKMES-1, H1703, Calu1 and H520 cells. Further studies included analysis of cell cycle, apoptosis, spheroids, migration and Pathscan intracellular signaling array. Expression of emerging proteins was validated by Western blot and evaluated by immunohistochemistry in tissue-microarrays from lung cancer patients. ResultsErlotinib and crizotinib showed additive interaction in Calu1, H520 and SKMES-1, and strong synergism in the LUDLU cells (Combination Index: 0.387), associated to G2/M phase arrest, increased apoptosis, spheroid size reduction and inhibition of migration. Remarkably, this combination decreased the phosphorylation of downstream targets of MAPK and PI3K/Akt/mTOR pathways, with the largest decrease observed for PRAS40 Thr246. Moreover, it reduced the expression of both p-Her3 and p-PRAS40 in the synergistic LUDLU cells. Tissue specimens showed a higher expression of both proteins in SCC compared to adenocarcinoma histology. ConclusionsCombining erlotinib and crizotinib led to an additive/synergistic interaction in 4 out of 5 SCC cells. By combining both inhibitors, MAPK and PI3K/Akt/mTOR pathways were strongly inhibited, leading to increased cell death. p-Her3 and p-PRAS40 might be used as markers for determining the synergistic effect and for selecting potential candidates for the combination treatment.

Immunohistochemical evaluation of cell cycle-related proteins in squamous cell carcinomas using digital image analysis: validation of the ImageJ immunoratio plugin

Immunohistochemical evaluation of cell cycle-related proteins in squamous cell carcinomas using digital image analysis: validation of the ImageJ immunoratio plugin

Immunohistochemical Expression of p53 and Bcl-2 in Algerian Cervical Carcinoma

The Objective of the present study is to evaluate the expression levels of Bcl-2 and p53 proteins in squamous cell carcinoma and adenocarcinoma of the uterine cervix, and try to explain their role as prognostic markers for this cancer. The cohort comprised 90 cases of the cervix lesions. The samples were assessed by immunohistochemistry for the expression of Bcl-2 and p53 proteins. The results showed that the Bcl-2 expression was either absent, low or moderate respectively in 38.96%; 50.65% and 10.39% of SCC cases. However, it was absent or expressed in 76.92% and 23.08% of adenocarcinoma cases respectively. The p53 protein was absent or present respectively in 75.32% and 24.68% of SCC cases as demonstrated by immunohistochemistry. p53 was almost absent in adenocarcinoma samples where only 7.70% of cases were positive. There was no significant correlation between Bcl-2 and p53 expression (p=0.352). We conclude that p53 expression, detected by immunohistochemistry, does not appear to be a prognostic marker for cervical cancer. Nevertheless, Bcl-2 expression seems to provide more information for this disease. It may represent an important indicator for cervical cancer.

Immunohistochemical Study Of Fibronectin, Tenascin and Type I Collagen In Tongue And Lip carcinomas

AIM: The aim was to compare the immunoexpression of extracellular matrix proteins in squamous cell carcinomas of tongue (SCCTo) and lower lip (SCCLi). METHODS: Eleven SCCTo and 11 SCCLi were selected and examined according to Bryne’s method (1998). For immunohistochemical study utilized antibodies to fibronectin, tenascin and type I collagen. Histopathologic and immunohistochemical analysis were performed on the tumor invasive front. RESULTS: All SCCTo were classified in high score malignant grade and all SCCLi in lower score. Fibronectin showed strong immunorreactivity in the peritumoral basement membrane (BM) in 91% of SCCTo and all cases of SCCLi, while in the tumor stroma (TS) all cases of SCCTo and SCCLi had strong intensity. Tenascin had strong expression in BM of 91% cases of SCCTo and 63.4% of SCCLi and in TS had strong expression in 91% cases of SCCTo and 54.6% of SCCLi. Type I collagen demonstrated weak immunoreactivity in the TS of 72.7% cases of SCCTo and 63.4% of SCCLi. CONCLUSION: These results may suggest that the strong expression of fibronectin and tenascin proteins and the weak expression of type I collagen could play a role in the invasive process of oral SCC.

Expression Profiling and Cellular Localization of Stress Responsive Proteins in Squamous Cell Carcinoma of Human Esophagus

ABSTRACTBackground: The ambiguity in relating expression dynamics of stress response proteins with human esophageal squamous cell carcinoma (ESCC) has sidelined the potential of stress proteins as therapeutic targets. This study was an attempt to unequivocally relate the stress protein dynamics with stage and propensity of ESCC. Methods: Surgically resected tumor and adjacent histologically normal tissue from 46 patients with esophageal squamous cell carcinoma were investigated in the present study. Expression of HSPs was analyzed by Western blotting and immunohistochemistry. Results: HSP expression was observed in all 46 cases both in adjacent normal and tumor tissues. The expression and the localization of individual HSP showed no significant correlation with depth of invasion, tumor grade, and pathological stage of the tumor. HSP 27 was the most abundant protein followed by HSP 90 and HSP 70. The HSP 27 localized exclusively in the cytoplasm of adjacent normal and tumor cells. HSP 70 showed dispersed expression with predominating nuclear localization in both normal and tumor tissue cells and HSP 90 was localized in cytoplasm of adjacent normal and in nucleus of tumor cells in majority of the cases. Conclusion: Our data advocate lack of relationship between stress protein expression and the progression of ESCC. The data renew the prospect of anti-HSP drugs as therapeutic resources in light of the possibility that their use would continue to sensitize cancer cells towards drug induced apoptosis for tumor regression.

Results of investigating the characteristics of bone marrow in patients with squamous cell carcinoma of the head and neck, their clinical value

To reveal the specific features of hematopoiesis in patients with head and neck squamous cell carcinoma (HNSCC), the authors analyzed the indicators of 41 bone marrow aspirates in relation to the clinical and morphological characteristics of the tumor and micrometastases detected in bone marrow. Puncture samples were morphologically and molecularly examined at the Laboratory of Hematopoiesis Immunology, N.N. Blokhin Russian Cancer Research Center. The found changes in the bone marrow cell composition in HNSCC patients showed themselves as an increase in the relative count of segmented neutrophils and a decrease in the level of immature granulocytes with no alternations in the neutrophil maturation index and in the percentage of granulocyte lineage cells. Bone marrow hypocellularity is a characteristic feature of high-grade HNSCC. The significantly enhanced leuko-erythroblastic ratio was due to a reduction in red blood cells: the erythroid cell maturation index rose, by increasing the percentage of bone marrow oxyphilic normoblasts in patients with locally advanced HNSCC. A highly sensitive method for detecting mRNA of the specific squamous cell carcinoma protein E48 was perfected using reverse-transcription polymerase chain reaction and Southern blotting, followed by an enzyme immunoassay that could recognize bone marrow micrometastases in 35 % of the patients with HNSCC. The patients with found bone marrow micrometastases showed a significant decrease in the proportion of polychromatophilic normoblasts and a significant increase in granulocytes in the presence of normal values of lymphocytes and other cellular elements of myelograms. It was established that in the presence of regional lymph node involvement, bone marrow micrometastases were significantly more frequently in the patients with HNSCC and tumor progression over a period of up to 9 months.

Expression and clinical implications of enhancer of Zeste homolog 2 and p53 protein in squamous cell carcinoma and precancerous lesions in the cervix.

We investigated the expression and clinical implications of enhancer of Zeste homolog 2 (EZH2) and p53 protein in cervical squamous cell carcinoma (SCC) and precancerous lesions. EZH2 and p53 expressions in SCC (168), cervical intraepithelial neoplasia (CIN)-I (19), CIN-II (35), and normal tissues (30) were detected by streptavidin-peroxidase-conjugation. The correlation between co-expression of EZH2 and p53 protein and the clinic pathological features and prognosis of SCC were discussed. The positive expression rates of EZH2 and p53 were 6.7, 37.0, and 75.6%, and 3.3, 21.1, and 39.3% in normal cervical tissues, CIN, and SCC, respectively, which were significantly different (P < 0.05). The positive expression rate of EZH2 and p53 protein in SCC patients with and without lymph node metastasis was 82.9 and 70.4% (EZH2) and 45.7 and 34.7% (p53), respectively, which was also a significant difference (P < 0.05). The progression-free survival (PFS) rates in followed-up patients (N = 143) who were EZH2- and p53-negative, EZH2- or p53-positive, and EZH2- and p53-positive were 71.3 ± 1.9, 66.1 ± 2.0, and 51.3 ± 3.8 months, respectively, which was a significant difference (P < 0.001); the overall survival among these groups was 72.9 ± 1.1, 68.6 ± 1.8, and 57.4 ± 3.4 months, respectively (P < 0.001). Multivariate analyses revealed that EZH2 expression, lymph node metastasis, and tumor staging were independent prognostic factors of SCC. EZH2 and p53, which affect lymph node metastasis and prognosis of SCC, may play a key role in the occurrence and development of SCC.

Expression of p53 and bcl2 in squamous cell carcinoma of head and neck

Background: Head and neck squamous cell carcinoma (HNSCC) is the sixth-most common malignancy worldwide. Despite advances in radiotherapy and surgical treatment, survival rates have not changed significantly in the last 40 years. Molecular markers are currently being identified that can determine prognosis preoperatively by routine tumor biopsy, lead- ing to improved management of HNSCC patients. Aim: The aim of the present study was to demonstrate the expression of p53 and bcl2 proteins in squamous cell carcinoma of head and neck (HNSCC) and to correlate the expression of p53 and bcl2 with clinical staging (AJCC) and WHO histological grading of SCC. Materials and methods: The study population comprised 50 cases of HNSCC. Tissue sections from these cases were subjected to hematoxylin and eosin (H&E) and immunohistochemical (IHC) staining using p53 and bcl2, and a comparative analysis of the results was performed. Cases of colon carcinoma and benign lymphoid hyperplasia were used as positive controls for p53 and bcl2, respectively. Results: Positivity for p53 was recorded in 30 cases (30/50), while positivity for bcl2 was recorded in 44 cases (44/50). Positivity for p53+/bcl2+ coexpression was seen in 28 cases (28/50). The frequency of p53 expression was associated with tumor histologic grade (p=0.02), increasing lymph node involvement (p=0.01), and clinical stage (p=0.038). The frequency of bcl2 expression was associated with histological grade (p=0.02) and increasing lymph node involvement (p=0.028), but not with clinical stage (p=0.242). Moreover, the combined p53+/bcl2+ expression was significantly associated with histo- logical grade (p=0.02) and lymph node involvement (p=0.01). Conclusion: Study of p53 and bcl2 expression may provide clinicians with more exact information in order to evaluate tumor aggressiveness and survival rates.

Glycosylated Alpha-1-acid glycoprotein 1 as a potential lung cancer serum biomarker

Presently existing screening approaches for lung cancer are not being proving sufficient and sensitive, so a study was conducted to identify disease related biomarker proteins for diagnostic applications. A total of 100 lung cancer patients (88 non-small cell lung cancer and 12 small cell lung cancer) and 50 healthy controls were included in this study. Serum samples of patients and healthy controls were subjected to a series of proteomic approaches and as a result of two dimensional gel electrophoresis, a ∼43kDa protein was found to be differentially expressed compared to healthy controls. Quantitative profiling of two dimensional gels by Dymension software analysis displayed 3.58 fold increased expression of ∼43kDa protein in squamous cell carcinoma and 2.92 fold in case of adenocarcinoma. Mass spectrometric analysis resulted in identification of 8 differentially expressed proteins, out of which human Alpha-1-acid glycoprotein 1 was targeted for further validations. This candidate protein exhibited N-linked glycosylation at five amino acid residues; 33, 56, 72, 93, and 103 with significant score of 0.66, 0.78, 0.78, 0.53 and 0.66, respectively. Sandwich ELISA quantified high serum levels of Alpha-1-acid glycoprotein 1 in squamous cell carcinoma (2.93g/l±1.22) and adenocarcinoma (2.39g/l±1.13) when compared with healthy controls (0.83g/l±0.21). One-way ANOVA analysis predicted highly significant variation of Alpha-1-acid glycoprotein 1, among all the study types (F-value 65.37, p-value 0.000). This study may prove as a non-invasive, cost effective and sensitive scheme for diagnosis of lung cancer, by passing the expensive and painful screening procedures.

Abstract 4477: A 5-nitrobenzoate-derived compound elicits anti-cancer metastatic activity by inhibition of poloplanin-stimulated tumor cell-induced platelet aggregation

Abstract Tumor cells dissemination and survival within the vascular system play an important role in cancer metastasis and is a clinical challenge for post-surgical cancer management. Podoplanin (PDPN) is one of the frequently over-expressed proteins in squamous cell carcinoma. By binding to platelet C-type lectin-like receptor 2 (CLEC-2), PDPN stimulates tumor cell-induced platelet aggregation (TCIPA) and facilitates cancer cell metastasis. In this study, a series of 5-nitrobenzoate-derived compounds were synthesized to screen for anti-cancer metastasis compounds that elicit functional blockage of PDPN-stimulated CLEC-2 activation and TCIPA. Platelet aggregation assays indicate that the small molecule compound 2CP selectively inhibited PDPN-induced platelet aggregation and the TCIPA of C6/Lung and osteosarcoma cells which expressed high levels of PDPN protein. Mouse tail bleeding time assay revealed that 2CP did not cause deleterious effects on normal blood hemostasis. In xenograft experimental metastasis model, 2CP possessed in vivo anti-cancer metastatic activity and augments the therapeutic efficacy of cisplatin; both the formation of pulmonary tumor foci and the body weight loss associated with the increased tumor burden were attenuated by combined treatment of 2CP and cisplatin. In conclusion, we present a novel and non-cytotoxic small molecule compound 2CP that elicits specific inhibitory effects on tumor cells-platelet interaction through intervening PDPN/CLEC-2 interaction and PDPN-mediated TCIPA. The findings are valuable for future application of 2CP in the prevention and treatment of cancer metastasis. Citation Format: Yao-Wen Chang, Pei-Wen Hsieh, Kowit-Yu Chong, Ching-Ping Tseng. A 5-nitrobenzoate-derived compound elicits anti-cancer metastatic activity by inhibition of poloplanin-stimulated tumor cell-induced platelet aggregation. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4477. doi:10.1158/1538-7445.AM2015-4477

Expression of Bcl-2, P16INK4a, and Cyclin D1 Proteins in Squamous Cell Carcinoma of the Lip

The protein expression of Bcl-2, p16INK4a, and cyclin D1 in squamous cell carcinoma of the lip (SCCL) was evaluated. Study Design: Sixteen cases of SCCL were studied by immunohistochemistry and evaluated according to histological grading and histological features. Results: The expression of p16INK4a was the highest among proteins studied, with 68.7% of positive cases, especially in well-differentiated SCCL and in keratin pearls, followed by Bcl-2 expression in 42.8% of cases, especially in moderately differentiated SCCL and in cords.

Abstract PR04: Distinctive squamous cell carcinoma protein signatures.

Abstract Squamous cell carcinoma (SQ) of the lung causes approximately 400,000 deaths per year worldwide, and no specifically targeted treatments yet exist. We used the SOMAscan proteomic platform (which measures 1129 proteins with a median limit of detection of 40 fM and 5% CV) in a broad-based analysis of serum and tissue samples to gain insight into the distinctive drivers of SQ malignant growth. We discovered unique SQ markers and pathways that show early expression of invasion and metastasis signals. Strong SQ signals were revealed. Our multi-center serum study of 94 non-small cell lung cancer (NSCLC) cases and 269 long-term smoker and benign pulmonary nodule controls resulted in the discovery of 60 lung cancer biomarkers and the development of a 7-marker diagnostic panel. This panel was validated in two independent cohorts. The AUC for detection of SQ carcinoma was 0.93 in training and 0.89 in the Univ. of Heidelberg validation set (56 SQ cases/27 benign nodule controls). Performance was confirmed with an AUC of 0.87 in an independent validation cohort assembled by the EDRN (25 SQ cases/20 benign nodules/52 smoker controls). This non-invasive test could be used as an adjunct to CT to detect rapidly growing SQ tumors that are disproportionately missed as interval cancers in CT screening studies. In addition, we analyzed 68 NSCLC tumor and matched non-tumor tissue lysates. This study consisted of 49 adenocarcinoma (AD) and 19 SQ tumors, 88% of which were Stage I or II. Proteomic comparisons of tumor/non-tumor or AD/SQ tissue samples performed using the Mann-Whitney test identified 79 tumor markers. Differences between tumor and non-tumor tissue were dominated by inflammatory, apoptotic and cell proliferation proteins. Just as we observed in the serum studies, the most common pattern was an increasing difference in protein levels from non tumor to AD to SQ. Of note, 24% of the proteins were only markers in SQ. These SQ-only markers are enriched for angiogenesis, cell proliferation and cell adhesion functions. Unexpected findings that the SQ proteome is dominated by proteins that promote malignancy and metastasis even in early stage tumors were revealed through broad proteomic profiling. These insights may lead to essential connections between the biochemical pathways altered in malignancy and tools for SQ cancer diagnosis and treatment. This abstract is also presented as Poster B14. Citation Format: Rachel Ostroff, Michael R. Mehan, Stephen Williams, Edward Brody, Harvey Pass, William Rom, Jill Siegfried, Thomas Muley, Wilbur Franklin, Dan Merrick, Adrie van Bokhoven, Holly Wolf, William Feser, Anna E. Barón, York Miller. Distinctive squamous cell carcinoma protein signatures. [abstract]. In: Proceedings of the AACR-IASLC Joint Conference on Molecular Origins of Lung Cancer; 2014 Jan 6-9; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2014;20(2Suppl):Abstract nr PR04.

Cisplatin benefit is predicted by immunohistochemical analysis of DNA repair proteins in squamous cell carcinoma but not adenocarcinoma: theranostic modeling by NSCLC constituent histological subclasses

Most non-small-cell lung cancer (NSCLC) patients receive cisplatin-based chemotherapy though clinical response is restricted to a subset of patients. DNA repair protein levels are possible surrogates for cisplatin-induced DNA adduct (and subsequent cell death) repair efficiency and thus molecular determinants of therapeutic efficacy. The International Adjuvant Lung Trial (IALT)-Bio study previously suggested ERCC1 and MSH2 as predictive of cisplatin-based therapeutic benefit. DNA repair protein expression (XPF, BRCA1, ERCC1, MSH2, p53, PARP1, and ATM) was assessed by immunohistochemistry on a large subset of patients (N = 769) from the IALT trial. Tissue Microarray slides were digitally scanned and signal quantified by user-defined macros. Statistical analyses (univariate and multivariate) of 5-year disease-free survival (DFS) and 5-year overall survival used binary cut-offs (H score low/high expression). In patients with squamous cell carcinoma (SCC), ATM, p53, PARP1, ERCC1, and MSH2 displayed significant (borderline) predictive values, mainly on DFS with chemotherapy efficacy limited to low marker levels. Adenocarcinoma (ADC) results were not significant. BRCA1 and XPF were not significant for predictive modeling in either SCC or ADCs. Here predictive utility of DNA repair enzymes co-segregates with SCC histology, focusing their predictive value to this histological subclass of NSCLC. Distinct mechanisms of chemotherapeutic response or resistance might exist among histological subclasses of solid tumors.