Esophageal squamous cell carcinoma (ESCC) is the dominant histological subtype in esophageal cancers (EC) and is one of the most aggressive cancers without effective therapy, Currently, Human papillomavirus (HPV) is suggested to be a potential risk factor for EC in addition to the classic predisposing factors, alcohol and tobacco, We have sought to investigate p16 a well-known biomarker for HPV and theranostic factors (EGFR & BRAF) that may have an impact on survival of ESCC. Methods We have analyzed 68 tissue biopsies which were histopathologically confirmed as ESCC. The paraffin embedded biopsies were subjected to immunohistochemistry (IHC) staining of EGFR and p16. The intensity of membrane staining for EGFR were scored as:0,1+,2+,3+, scores 2+ and 3+ were taken as over expression and score 1 as low expression. IHC for p16 was observed positive if it was brown colour in nucleus and cytoplasm. Then, mutational analysis for BRAF V600E was performed on a non biased data base by using PCR technique and confirming it through bidirectional Sanger sequencing. Results In total of 68 biopsies: Immunohistochemistry, - 56 biopsies (82.3%) overexpressed EGFR of which high grade ESCC’s showed greater expression than low grade. - 42 biopsies (61.7%) expressed p16 - 23 biopsies (33.8%) showing positivity for both p16 & EGFR thereby explaining p16 mediated EGFR upregulation Mutational analysis: Two middle aged female patients showed mutants: - Mutation of ACAGAAAAA was detected in V600E codon for one - Mutation of TCTGTATCA was detected in exon 11 of G466V for another - However IHC for p16 & EGFR of these two patients turned negative. Conclusion Our study reveals comprehensive mutational profile (EGFR, p16 and BRAF) on ESCC, shedding light on potential molecular mechanisms associated with its development and possible therapeutic regimens. Taken together with our findings, we propose a prospective study of a large sized population especially including middle aged female patients to further validate the role of this profile as indicators to unravel the theranostic values. To our knowledge this study is the first of it’s kind in India.