EVALUATION OF IMMUNOHISTOCHEMISTRY OF TP53 GENE WITH RISK OF SQUAMOUS CELL CARCINOMA HEAD AND NECK (HNSCC) IN KASHMIRI POPULATION

Dr Shakeel Mohmad Wani 1 , Dr Shahnaz Sheikh 1 , Dr Parth Jigishbhai Amin 2 , Professor(Dr) Ruby Reshi 3 and Dr Umeek Jeelani 2 . 1. Resident ENT HNS, SMHS Hospital, Srinagar, Kashmir, India. 2. Resident ENT, MCH SKIMS, Srinagar, Kashmir, India. 3. Head of Department Pathology Govt. Medical College, Srinagar, Kashmir, India. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History


Objective:-
To find relationship between varied Immunohistochemistry of TP53 and squamous cell carcinoma head and neck (SCCHN) . Study design:-Case control study.

…………………………………………………………………………………………………….... Background:-
The incidence of squamous-cell carcinoma continues to rise around the world. A recent study estimated that there are between 180,000 to 400,000 cases of SCC in the United States in 2013. 1 Risk factors for squamous-cell carcinoma varies with age, gender, race, geography, and genetics. The incidence of SCC increases with age and the peak incidence is usually around 60 years old. Males are affected with SCC at a ratio of 2:1 in comparison to females. Caucasians are more likely to be affected, especially those with fair Celtic skin and chronically exposed to UV radiation. Squamous-cell carcinoma of the skin is the most common among all sites of the body. Solid organ transplant recipients (heart, lung, liver, pancreas, among others) are also at a heightened risk of developing aggressive, high-risk SCC. There are also a few rare congenital diseases predisposed to cutaneous malignancy. In certain geographic locations, exposure to arsenic in well water or from industrial sources may significantly increase the risk of SCC.
Ninety percent of cases of head and neck cancer (cancer of the mouth, nasal cavity, nasopharynx, throat and associated structures) are due to squamous cell carcinoma. Symptoms may include a poorly healing mouth ulcer, a hoarse voice or other persistent problems in the area. Treatment is usually with surgery (which may be extensive) and radiotherapy. Risk factors include smoking, alcohol consumption and hematopoietic stem cell transplantation 2 Aim of the study:-To find relationship between squamous cell carcinoma head and neck (SCCHN) and its impact on p53 protein expression.

Corresponding Author:-Dr Shakeel Mohmad Wani.
Address:-Resident ENT HNS, SMHS Hospital , Srinagar, kashmir, India. Immuno-histochemistry for tp53 protein expression:-Procedure:-The wax blocks of cases and controls were de-paraffinized with xylene 2 changes lasting 5 minutes each. Hydration was done using 100% ethanol 2 changes for 3 minutes each followed by 95% and 80% ethanol for 1 minute each. Then rinsing was done with distilled water. Pre-heating was done using Pre-heat steamer or water bath with staining dish containing Sodium Citrate Buffer or Citrate until temperature reaches 95-100OC .The slides were immersed in the staining dish and incubated for 20-40 minutes. After Turning off steamer slides were removed and allowed to cool for 20 minutes. Sections were rinsed with PBS Tween 20 for 2x2 min. Sections were incubated with primary antibody at appropriate dilution in primary antibody dilution buffer for 1 hour at room temperature or overnight .Then sections were rinsed with PBS Tween 20 for 2x2 min and blocking was done with peroxidase blocking solution for 10 minutes. Again rinsing was done with PBS Tween 20 for 3x2 min.

Materials and Methods:-
1 drop of freshly prepared liquid DAB Chromogen in 1 ml stable DAB buffer was added to the tissue and incubated for 5 minutes at room temp(20-25 O C). Hematoxylin counter staining was done and incubated for 3 min at room temp ((20-25 0 C).The slides were mounted and studied for TP53 positivity.