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Esophageal cancer is the 8th most common type of cancer and the 6th most common cause of death from cancer, globally.Human papillomavirus (HPV) is a non-enveloped, double-stranded DNA virus with more than 200 genotypes.To date,molecular and epidemiological studies repeatedly demonstrated the relation between cervical cancer and HPV virus and it has recently also been established as a risk factor for Oro-pharyngeal squamous cell carcinoma.Esophageal squamous cell carcinoma (ESCC) has a distinct geographic distribution with a high prevalence in certain regions of the world, the most common carcinoma in China, South Africa and some other areas.The etiology of ESCC still remains unclear, but it is most likely multifactorial.The role of human papillomavirus (HPV) in the etiology of ESCC has been debated over past 20 years.The aim of this research is to evaluate the presence and relationship between HPV, especially high risk HPV16 & HPV 18, and ESCC in vitro.Detection of HPV relies strictly on molecular analyses of HPV DNA sequence.In clinically resected esophageal samples HPV was detected frequently through polymerase chain reaction (PCR) and in situ hybridization (ISH).The incidence of infection of HPV in ESCC, however, differs markedly depending on the geographic location of patient population being studied.Frequency of detection of HPV genome in cancerous tissues varies within different areas from 0％ to 71％.The highest reported prevalence of HPV occurs in the high-risk regions for ESCC in China and South Africa.In this study we launched an evaluation on 195 paraffin embedded tissue blockes we got from pathology department of Tang Shah area of China, as a high incidence area for ESCC, from 2008 to 2012.PCR results after evaluation of HPV16 and HPV18, high risk HPV virus, showed high percentage of genome existence of GP5+/GP6+ (79.79％), HPV16 (40.40％) and HPV18 (47.47％) in cancerous tissues.For the confirmation of the PCR results and to use another complementary method to check HPV genome presence and expression we chose Immunohistochemistry (IHC) method to investigate the proteins related to HPV virus and a few common cancerous proteins like P21 and P16, common biomarkers in detection cancer.The results of this step also confirmed the previous step of evaluation with relatively high prevalence of 41.9％ for E6HPV16/18, 22.7％ for E7HPV16/18, 41.9％, 38.4％ for P16 and 37.9％ for p21.At next step for detection of carcinogenesis of high risk HPV virus in ESCC we used new born blank esophagus tissue and after cell extraction, Established human esophagus epithelium (SHEE), transfeeted this cell line by HPV virus and checked the carcinogenesis of virus after many times culture in different generations, 24, 48,60(1827), 76 with high sensitive gene screening method, DNA microarray.The aim of this step of our research firstly was confirming the effect of HPV infection in the genome of cell and consequent carcinogenic action and the second aid was finding upper-regulated genes after infection during carcinogenic procedure.After analyzing results of this step we found four genes with high expression after HPV infection,AIM2, TBGF2, MAGE A3 and MAGE A6 that one of them after IHC evaluation on ESCC cancerous tissues slides showed direct relation with positive HPV PCR result,MAGE A6.This study found a presence of HPV DNA in esophagus tumors (high risk HPV types 16 and 18) implicates HPV as one of the possible etiology in esophageal squamous cell carcinoma.On the other hand we presented one probable related gene to ESCC caused with HPV infection and this achievement can give new ideas for other researcher to start new study plans with the aim of finding cure, vaccine or detection way of this kind of cancer, ESCC.