|Year : 2016 | Volume
| Issue : 3 | Page : 1-4
Basic Science Research
|Date of Web Publication||27-Sep-2016|
Source of Support: None, Conflict of Interest: None
|How to cite this article:|
. Basic Science Research. J Head Neck Physicians Surg 2016;4, Suppl S1:1-4
BS1: Selected Molecular Markers as Indicators of Clinical Profile, Tumour Characteristics and Treatment Outcome in Squamous Cell Carcinoma of the Larynx
Elizabeth Mathew Iype, B Rajan, S Lakshmi,
V T Jissa, K Jayasree, Philip John
Regional Cancer Centre, Trivandrum, Kerala, India
Loco-regional recurrence in Laryngeal carcinoma still poses a significant oncological management dilemma for a subset of these tumours. Therefore, the possibility that specific tumour markers may aid the clinician in choosing optimal treatment for a specific tumour holds great promise. Objectives were to study the expression of molecular markers p53, Bcl-2, EGFR, Ki67, Cyclin D1 and Cox-2 in LSCC so as to form a hypothesis that a particular group of markers will be useful in determining the prognosis. We studied 72 patients with LSCC in an attempt to determine relationship between their clinicopathological characteristics and treatment outcome with the expression of these six molecular markers. This is one of the largest comprehensive studies of multiple molecular markers in predicting ECS, PNI, LN spread and survival. EGFR, Cyclin D1, and Ki67 were intense in ECS and PNI. N0 with intense Cyclin D1 or Ki67 had worse DFS. Old age and T4 stage with intense p53, Bcl-2 had worse outcome. Cox-2 was highly predictive of Nodes. In advanced carcinoma larynx cases, EGFR, Cyclin D1, and Ki67 estimation will be beneficial in planning treatment and should form the integral part of work-up. N0 cases with intense cyclin D1 or Ki67 would do better with prophylactic neck treatment. P53/Bcl-2 should be determined for prognostication in old age and T4 stage. Cox-2 will be useful in predicting the occult nodes. From our study, we categorized the 6 markers in to three groups as markers of aggressiveness, markers of invasiveness and markers predicting survival.
BS2: To Study the Varied Histomorphology of all Sinonasal and Nasopharyngeal Masses and Unusual Lesions in these Locations
Tanushri Mukherjee, Ravi Roy, Rajat Dutta,
Command Hospital, Kolkata, West Bengal, India
Background: Sinonasal tract and nasopharyngeal tract masses have diverse morphology and histopathology due to anatomic and embryonic distinction with occurrence of unusual lesions or tumors in these location. Methods: A retrospective observational study for 5 years from 2011-2016 till date (May 2016) in Command hospital was carried out. Total analysis of 125 cases was done to study the varied clinical presentation and histomorphology of all sinonasal and nasopharyngeal masses and also note the unusual types of lesion rare in this location. Results and Discussion: This is a retrospective analysis of 125 cases of sinonasal and nasopharyngeal masses. Clinical examination and radiological findings were correlated with histomorphology. Out of total 126 cases, 42 (33.3%) were non-tumorous and 84 (66.6%) were neoplastic masses, 23 masses (18.2%), were proved to be of malignant etiology. In benign neoplastic category unusual entity were Chordoma, hemangiopericytoma, meningioma, cavernous hemangioma. Amongst the malignant cases the distinct and unusual entities were diffuse large B cell lymphoma, esthesioneuroblastoma, sinonasal adenocarcinoma, adenoid cystic carcinoma. Conclusion: Sinonasal and nasopharyngeal masses may arise in distinct locations and are non-neoplastic and neoplastic with benign and malignant varied histopathology and also the unusual varieties also occur which rare to these locations and should be promptly diagnosed for accurate treatment.
BS3: Molecular Predictors of Chemotherapy Resistance/Response in Head and Neck Cancer
Ram Bhupal Reddy, D R Ravindra, Naveen Hedne, Vikram Kekatpure, Moni Abraham Kuriakose, Amritha Suresh
Mazumdar-Shaw Centre for Translational Research, Integrated Head and Neck Oncology Program, Bengaluru, Karnataka, India
Objectives: The study aims to develop head and neck squamous cell carcinoma (HNSCC) specific panel of biomarkers of resistance/response to standard chemotherapy drugs (Taxol and Platinum) using a systematic meta-analysis of existing databases. Methodology: Gene expression databases of squamous cell carcinoma stratifying response/resistance to platinum or taxol were obtained from the public domains. This data was analyzed using Gene spring software [v12.5, Agilent, USA] and cross compared with HNSCC database previously developed by our group (1). Quality control evaluation and statistical analysis were carried out as per analytical pipeline. The statistically significant genes (p-value < 0.05 and fold value >2) were then validated in patients by expression profiling at transcript and protein levels in three patient cohorts [Group I: Surgery followed by adjuvant chemotherapy with radiotherapy; Group II: Neo-adjuvant chemotherapy; Group III: Recurrent patients]. PET-CT evaluation was carried out for treatment response assessment. To determine biologic significance, small molecule inhibitors targeting selected markers were used in HNSCC cell lines. Results: Microarray series (n = 9) that profiled response to taxol/platinum therapy in squamous epithelial cancers were analyzed. These included 133 Non-responders (NR) and 55 responders (R). 78 entities were identified after cross comparison with HNSCC database (Platinum = 65; Taxol = 8; common = 5). Among this, a set of 17 genes were validated by qPCR in patient group I (N = 40). ROC curve analysis revealed set of 7 markers (MMP2, SPP1, FZD6, SNAI2, COL3A1, SLC2A1 and CEP55) with high predictive value (AUC > 0.5). The top markers (MMP2, CEP55 and FZD6) are being validated by IHC. The role of these markers in resistance to chemotherapy will be evaluated using specific inhibitors in Cisplatin resistant cell lines (CAL-27 CisR) (2) by cytotoxicity assay and profiling of downstream markers. A similar experimental pipeline will also be used for identifying markers relevant to Group II and III. Conclusion: The study is an attempt to develop a panel of markers for predicting resistance/response markers in different patient cohorts of HNSCC and to understand the molecular mechanism of resistance response to the treatment. The study also attempts to identify target molecules to reverse the resistance to the therapy.
BS4: Molecular Analysis of Drug Treated Oral Cancer Cell Line Cal27 by LC-MS/MS Mass Spectrometry and Bioinformatics
Priya Sivadasan, N Reddy Harsha Vardhan,
Manoj Kumar Gupta, Safeena Kulsum,
Amritha Suresh, Moni Abraham Kuriakose,
Mazumdar Shaw Cancer Centre, Bengaluru, Karnataka, India
Background: Though induction chemotherapy has demonstrated over 60% response among patients, disease relapse is a common feature. This modality also has not lead to improvement in survival rate. Combination of Taxol, Platinum and 5-fluoro uracil (TPF) is the standard regimen for HNSCC patients as induction chemotherapy. Inherent or acquired resistance to these drugs in a subset of patients contributes to treatment failure. The aim of this study is to understand the molecular changes in the drug treated OSCC cell line Cal27 to understand factors that may be responsible for chemo-resistance and would help in treatment management. Methods: Differential protein expression profiling of the parental cell line (Cal 27 P) and its drug resistant versions (developed in the lab, Kulsum et al., 2016, Sindhu et al., 2015) - Cisplatin (CisR), 5FU (5FUR) and Cisplatin + 5FU + Docetaxel (TPFR) were carried out by iTRAQ - based quantitative proteomics analysis. Functional classification and biological pathway analysis of the differentially expressed proteins (DEPs) was carried out using Uniprot and KEGG database. Results and Discussion: There were 229 DEPs observed in CisR; (215 up and 14 down) 113 in 5FUR (63 up, and 50down) and 511 in TPFR (376 up, 135 down). 112 of these DEPs found to be present in patient tissues based on comparison with our database of DEPs in OSCC. These DEPs were also compared with the IHC data on head and neck cancer from HPA to confirm its presence in tissue and we found that 150, 83 and 364 DEPs from CisR, 5FUR and TPFR, respectively to overlap. The major biological processes represented were of protein metabolism, apoptosis and actin cytoskeletal reorganization. The major biological pathways represented were Ribosome, metabolic any particular, RNA transport and protein processing in endoplasmic reticulum. We found that two proteins COPA and HIST1H1B to be common DEPs for all the drug resistant cell lines. COPA is known to be associated with inhibition of apoptosis in mesothelioma and HIST1H1B is associated with DNA methylation. Conclusion: We identified DEPs in CisR, 5FUR, TPFR cells of Cal27 cell line which are detectible in HNSCC tissue and in saliva. Ribosome, metabolic and ECM-receptor interaction pathways are found to be deregulated in drug resistant cells compared to parental cells. The proteins COPA and HIST1H1B need to be further explored to understand its role in OSCC chemo resistance.
BS5: Application of Nanotechnology for Targeted Imaging of Sentinel Lymph Node
Mohammad Faiyaz Anwar, Sarat K Kottarath,
Amit K Dinda, Madhusudan Bhat, Alok Thakar
All India Institute of Medical Sciences, New Delhi, India
Background/Hypothesis: Detection of sentinel lymph node is an important procedure for accurate prognostication as well as management of the head and neck cancer. Conventional method of injection of dye and radioactive material for detection is not specific as well as involves the time consuming method of microscopic confirmation on frozen section. Methods: We fabricatedthree different nanoparticles composed of calcium phosphate, organically modified silica (ORMOSIL) and low molecular weight collagen. The FDA approved fluorescent dye for surgical use Indocyanin green (ICG) was loaded in these particles. Maximum fluorescence efficiency of ICG was achieved with collagen nanoparticles. These dye loaded nanoparticles were bioconjugated with cytokeratins via EDC and NHS method which are thoroughly characterized using analytical and spectroscopic techniques. Results and Discussion: TEM showed the particles with diameters in the range of 100-150 nm with a core and shell structure. The efficiency of specific binding of the ICG loaded cytokeratine labeled nanoparticles was demonstrated on the frozen section of metastatic lymph node. The particles bound to the cancer epithelial cells with specific fluorescence with out any nonspecific binding to the lymphoid tissue. Conclusion: Thus this study showed the translational potential of the nanoparticle system for specific targeted imaging of sentinel lymph node which may replace the nonspecific time consuming conventional method. (Acknowledgement: Dept. of Biotechnology, Govt. of India for financial assistance).
Key words: Cytokeratins, fluorophore, glutathione, stabilized collagen nanoparticles, tumor surgery
BS228: Recurrent Laryngeal Nerve: Anatomy Revisited in Indian Population
Nitika Gupta, Rohan Gupta, Sonika Kanotra,
Goverment Medical College, Srinagar, Jammu and Kashmir, India
Aim: To study RLN and its branching pattern relation with ITA. Methods: In our observational study done on 60 cadavers in mortuary and 50 patients undergoing thyroid surgeries 194 recurrent laryngeal nerves were studied (100 on right and 94 on left) for its anatomical course and relations with other structures. Results: Extralaryngeal branching of the nerve was seen in 50% of the nerves (51% on right and 48.93% on left). Cervical course of the nerve was divided in four segments in which the neuro-vascular crossing (the RLN and the ITA) and laryngeal entry points were taken as important landmarks for regional classification and location of terminal division of the nerve. The most common type of division on right was type 4 (prearterial) seen in 41.18%, followed by type 3 (prelaryngeal) seen in 27.45%, type 1 (arterial)[21.57%] and type 2 (postarterial)[9.8%]. The most common on left was type 3 in 39.13%, followed by type 4 [23.91%], type 1 [21.74%] and type 2 [15.22%]. In similar study by Gurleyik et al. arterial and postarterial division patterns were common which is not true for Indian population. Conclusion : In mobilization of the gland, the relationship of the nerve and the artery is important while placing ligature in the inferior thyroid artery. In Indian population the nerve and its branches are mostly at risk at the nerve entry and even before crossing of inferior thyroid artery thus they must be identified and preserved to prevent nerve injury.
BS229: Prognostic Evaluation of MCL-1 and its Stabalizing Factor TCTP in Head and Neck Squamous Carcinoma Treated with Chemoradiotherapy
Manish Mair, Tanuja Teni, Prasad Sulakshene, Vedang Murthy
Tata Memorial Hospital, Mumbai, Maharashtra, India
Background: In order to predict and improve the therapeutic outcome, identification of potential clinically relevant molecular markers for early diagnosis and optimal treatment planning are necessary. Evasion of apoptosis is an important hallmark of tumor cells which confers them the ability to sustain cell viability under stressful circumstances. In this study, we look into few of these molecular markers as a predictor of prognosis for head and neck cancer. Methods: This study includes 110 Squamous cell carcinoma patients (oropharynx, hypopharynx and larynx) with tumour stage T1-T3 and nodal stage N0-N2. These patients were treated primarily by Concurrent chemo radiation (CCRT) or Radiation (RT) alone. Follow up detail were prospectively maintained and evaluation of patients was done using PETCECT scan. Immunohistochemistry (IHC) for p16, MCL-1 (Myeloid cell leukemia1) and TCTP (translationally controlled tumour protein) was done on five micron thick sections from FFPE blocks. Results: Median age of the patients was 55 years with male to female ratio of 3.5:1. Node positivity was seen in 57.5% patients. MCL-1, TCTP expression was significantly less in poorly differentiated carcinoma (p - 0.000) and had no association with nodal positivity (p - 0.512). We found significant correlation of TCTP expression with MCL-1 expression (p - 0.01/r - 0.621). Median follow up period was 35 months. Eleven patients died due to other causes (10%), 21 (19.1%) died due to disease, 14 (12.7%) were alive with disease and 64 (58.1%) patients were alive without disease. MCL-1 (>50%) overexpression was associated with statistically significant poor survival. We found no association of TCTP with survival. The patients with p16 positivity and not habitual to tobacco had better survival outcomes. Conclusion: TCTP stabilizes MCL-1 protein and overexpression of MCL-1 protein can be used as a poor prognostic marker for head and neck cancers treated primarily by RT or CCRT. Overexpression of P-16 predicts better survival.
BS230: Targeting Aldh1a1 as a Chemotherapeutic Adjunct in Oral Cancer
Safeena Kulsum, H V Sudheendra, D R Ravindra, Gangotri Siddappa, R Nisheena, Amritha Suresh, Moni Abraham Kuriakose
Mazumdar Shaw Medical Foundation, Bengaluru, Karnataka, India
Objective: The objective of this study is to evaluate the relevance of ALDH1A1 in chemoresistance and the efficacy of its targeting towards improving the efficacy of chemotherapy in Oral squamous carcinoma. Methodology: Aldh1a1 expression in the treatment naive (Group I) and post chemotherapy (Group II) patient were evaluated using Immunohistochemistry (IHC) to correlation its expression with treatment. Expression pattern in resistant cell line, Cal-27 CisR, enriched with high MDR genes and Aldh1a1 positive cells were targeted using Aldh1a1 inhibitors (Si-RNA based targeting and small molecule inhibitor (NCT-501)) to evaluate effect on tumourigenicity, resistance and migration. Expression profile (qPCR) and functional assays (spheroid formation and migration assays) were used to confirm the role of Aldh1a1 inhibition on CSC mediated self-renewal (tumourigenic), chemoresistance and migration. Cell viability (MTT) assay was carried out to evaluate IC50 of Aldh1a1 silenced cells. In an ongoing experiment we try to confirm definite role of Aldh1a1 in chemoresistance, migration and chemotherapeutic response by increasing the Aldh1a1 expression exogenously using GFP tagged-ORF plasmid vector system in Cal-27 and Cal-27 CisR cell lines (in vitro). Finally, the clinical relevance of targeting Aldh1a1 with Cisplatin was evaluated in Cal-27 CisR derived xenografts in Hsd Athymic Nude-Foxn1 nu female mice (in vivo). The effect of targeting Adh1a1 expression was also assessed in oral cavity surgical tissue samples by ex vivo treatment using cisplatin, NCT-501 and combination. Results: Immunohistochemisrty analysis showed a significant increase in Aldh1a1 expression (p = 0.034) among the Group II patients (230 ± 20). Cytotoxicity assay showed 30-35% reduced cell viability in si-RNA silenced cells and 16% reduction in NCT-501 based Aldh1a1 inhibited cells along with a down regulation of MDR genes MRP2 (p = 0.001), ERCC1 (p = 0.043), CTR1 (p = 0.14) as compare to Cal-27 CisR cells. Similarly, functional assays showed reduced migration (p = 0.001) with 13% (p = 0.008) and 32% at 18 hours and 24 hours respectively as compared to 26% and 41% of the untreated control. Reduced spheroid formation capacity in NCT-501 based Aldh1a1 inhibition 40 nM (p = 0.0001) and 80 nM (p = 0.0001) in number and size 40 nM (0.0001), 80 nM (0.0001) were also observed. NCT-501 in combination with Cisplatin in in vivo study showed >50% reduction in tumourigenicity (213 mm 3 ) (p = 0.0009) as compared to untreated (512 mm 3 ). Post treatment response to relapse and recurrence are under observation. Ex vivo explant study (n = 4), with 5 uM cisplatin and 20 nM NCT-501 treatment showed a significant decrease in Ki-67 index (25.2% ± 1.18, p = 0.0015) as compared to untreated or cisplatin treatment ex vivo. The effect of exogenous expression of ALDH1A1 in the cells is currently in progress. Conclusion: Aldh1a1 targeting induced sensitivity to cisplatin in in vitro, in vivo and ex vivo models of oral cancer with possible therapeutic implications as an adjunct to chemotherapy.
BS231: Compliance with Treatment Recommendations in Oral Cancer: A Single Institution Experience
Vikram Bhardwaj, Shamit Chopra,
Anubha Bharthuar, Tanya Sharma
Patel Hospital, Noida, Uttar Pradesh, India
Oral cavity cancers are the commonest subsite among head and neck cancers. A significant number of patients diagnosed with Oral cavity cancers do not complete treatment owing to multiple factors. Treatment compliance is crucial for optimal treatment outcomes. Objectives: To assess compliance in a cohort of oral cancer patients in a single tertiary level cancer institute. To attempt correlation of compliance with multiple variables. Materials and Methods: A retrospective analysis of patients with elicitable data and all stage of oral cancer presenting to head and neck OPD of a single institute in tier 2 city was performed. Factors analyzed: age, gender, co-morbidities, stage, compliance for primary vs recurrent disease, relapse and type. Pearson Chi squared test was used to identify significance of observed values. Results: 367 patients were identified of which 290 were primary and 77 were recurrent disease. Overall compliance to treatment recommended in the first line setting was 59.4% (n = 218) while compliance for recurrent/relapsed disease was 79.1% which was statistically significant (p = 0.01). Stage I/II patients had significantly higher compliance compared with Stage III/IV (p = 0.01). Patients with comorbidities had higher compliance as compared to those with no comorbidities (p = 0.009). Compliance was higher when surgery as a treatment modality was offered (p = 0.03). No significant disparity was noticed in compliance rates based on age or gender.
BS232: Co-relation of p53, p16 and Cyclin D Expression in Oral Cavity Squamous Cell Carcinoma with Tumour T, N Stage and Grade
Lourembam Sunil Singh, Harit Chaturvedi, Bishwajyoti Hazarika, Vikas Singh, Javeed Altaf, Bikram Deka
Max Super Specialty Hospital, Saket, New Delhi, India
Introduction: Expression of certain proteins in squamous cell carcinoma of oral cavity have recently been explored in attempt to stratify them into prognostically different types. Such proteins expression includes p53, p16 and Cyclin D. Aim: to asseses any relation of p53, cyclin D and p16 protein with T, N stage and tumour grade in Oral cavity squamous cell cancers. Methods: We retrospectively collected 46 cases of squamous cell carcinoma of oral cavity, operated at Max Super specialty Hospital, Saket between Jan 2014 to June 2016. No neoadjuvant treatment were given in all cases. Expression of p53, p16 and Cyclin D protein were assessed using IHC and their expression were co-related with their T stage, N stage and tumour grade. Results: Expression of p53 and Cyclin D protein were related with higher tumour stage and poorer differentiation. Interestingly, expressions of p53 were very poor in our study cohort. Conclusion: p53 and cyclin D expression associated with higher T and N stage and poorer differentiation.
BS233: Is Elective Neck Dissection is Necessary in Early Tongue Carcinoma?
Vikas Singh, Harit Chaturvedi, Biswajyoti Hazarika, Sunil Singh, Javed Altaf, Bikram Deka
Max Super Speciality Hospital, New Delhi, India
Introduction: The indication of neck dissection in tongue carcinoma is a problem of risk - benefit evaluation b/w probability of neck metastasis and the problem of complication a/s with neck dissection. There is no consensus on the elective treatment of the neck in early oral cancer patients with a clinically N0 node. Whether patient with early stage cancer should be treated with elective neck dissection at the primary surgery or with therapeutic neck dissection after nodal relapse during watchful waiting. It is proposed that occult metastasis are detected in approximately 30% of all N0 patients and it is associated with higher rates of relapse rates in N0 patients on watchful waiting. Patients and Methods: We retrospectively analysed 150 patients of anterior tongue cancer with n0 nodes treated at max cancer institute delhi between 2010 t0 2016. All patients were treated with partial glossectomy elective modified radical neck dissection. Histopathology report of all patients are examined for occult and skip metastasis and pattern of nodal metstasis. Reults: The overall micrometastasis rate in our patients favours watchful waiting with therapeutic neck dissection on relapse. Moreover it was the incidence of metastases to level IV from early carcinoma is very low and not in favour of extended supraomohyoid neck dissection.
BS351: Nanotechnology: Revolution in diagnosis and treatment of cancer
Priyanka Raina, Kaustubh Patel
HCG Cancer Centre, Ahmedabad, India
Nanotechnology is an area of science devoted to the construction of molecular structures in the nanometer scale size range (often 100nm or smaller) . It derives its name from Greek word for "dwarf". Nanotechnology can target a tumor, carry imaging capability to document the presence of tumor, sense pathophysiological defects in tumor cells, deliver therapeutic genes or drugs based on tumor characteristics, respond to external triggers to release the agent and document the tumor response and identify residual tumor cells. Different kinds of nanoparticles suitable for drug and gene delivery, probing DNA structures include Liposomes, Polymeric Nanoparticles (Nano spheres and Nano capsules), Solid lipid particles, Nano crystals, Polymer Therapeutics such as dendrimers, fullerenes, Inorganic Nanoparticles eg. Gold & Magnetic Nano particles.The major area in which nanomedicine is being developed in cancer involves:
- Early detection of tumours (analysis of cancer associated markers and designing contrast agents that improve the resolution of tumour area comparing with the nearby normal tissues).
- Cancer treatment (Creating nanodevices that can release chemotherapeutic agents).
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