The Research About Human Papilloma Virus (HPV) Infection And P16Gene Mutation In Esophageal Squamous Cell Carcinoma

Background: The carcinoma of the esophagus is one of the mostcommon malignancies in China with high mortality. The prevalence in southof Hebei province, the Taihang mountains and north of Henan provinceregions is the highest of the word. The mainly histological types is esophagealsquamous cell carcinoma (95%). The early symptoms of this disease are notobvious, and easily overlooked. An early diagnosis and appropriate treatmentlead a better prognosis. Most of the patients are in advanced stage whendiagnosed, which is the reason of poor prognosis. So the prevention ofetiology early diagnosis and treatment are very important to improveprognosis. Human papilloma virus (HPV) was demonstrated to associate withmany tumour since it was found at the beginning of the20th century. Forexample, Human papillomavirus(HPV)infection is necessary for thedevelopment for cervical cancer. The success of HPV prophylactic vaccinedevelopment is the milestone of cervical cancer prevention of humankind.Syrjanen complete the first research on the link between HPV and ESCC in1982. Some scholars like Zhoujian did the same research for the first time inChina in1987. To work out the relationship between HPV and ESCC, muchresearch has been done at home and abroad. The rasearches found that insome area the HPV might be the reason of ESCC. But in another area theresult might contrary. Even in the same aera the different methods andresearchers could get different results. Therefore, the relationship betweenHPV infection and ESCC has still not been confirmed. p16gene, a newmember of the tumor suppressor genes in people, is the first one which canacts at the cell cycle. It plays a more and more important role in preventingcancer. It is known that p16gene change in tumour cell. But what is the reasonof the change? Is HPV infection the real reason for the change in ESCC? Objectives: This study use application polymerase chain reaction (PCR)joint DNA probe technologies to detect the satuation of HPV infection inESCC in south centre area of Hebei and further provide basis for vaccinedevelopment. The reasearch also use Polymerase chain reaction(PCR)-singlestrand conformation polymorphism (sscp) to screen the esophageal cancerand control groups for finding the change of p16gene exon2and to work outthe change in the ESCC process and the relationship between HPV infectionand ESCC.Methods: The tissue samples came from two groups. Esophageal SCCsamples were obtained from surgery specimens of47patients from southernarea of Hebei province. In the control group esophageal micosa were obtainedendoscopically from37healthy volunteers who came from the city ofShijiazhuang. QIAamp DNA mini kit was used to extract DNA. And HPVwas detected with LINEAR ARRAY HPV Genotyping Test kit of the DNA.β-Globin was used as internal control to ensure DNA quality. In addition tothe positive and negative control of the kit, HPV positive cervical cancertissres were used as the positive control and DEPC water, the negativecontrol. HPV DNA was detected using HPV E6gene primer p16genesexon2primers was used to PCR (design the primer to three parts to enhancesensitiveness), and join degeneration into PCR products with high temperatureto degeneration for5minutes.The DNA electrophoresis of polyacrylamide gelwas carried out in4℃. After the electrophoresis,silver stain method wasused to view the expression. Statistical analysis was conducted using SPSS13.0and SAS8.0.Results:1The infection rate of HPV was0%in esophageal squamous cell carcinomapatients,0%in the control group, positive in the positive control of the kit,positive in the cervical specimen, HPV E6gene expression was0%inesophageal cancer, and0%in the control group, positivein the cervical cancersamples2P16exon2gene Deletion was0%in control group, the mutation rate is 44.7%. p16exon2gene Deletion was0%, the mutation rate was0%in controlgroup. Rank-sum test was used to detect the change of p16gene in thedifferent pathological stage The results showed that the asymptotic significantvalue was0.002, chi-square value was12.738. The results showed that thelater pathology stages, the higher p16gene exon2variation.3Using Pearson related analysis, the gradual significant bewteen p16genesexon2of patients' age, gender, smoking history, drinking history, familyhistory and lymph node metastasis was:0.496,0.143,0.000,0.547,0.113,0.005.4To detect the relationship between p16gene exon2and HPV infection,Fisher exactly methods analysis showed that p>0.05.Conclusion:1In the south centre of hebei province in China, the esophageal squamous cellcarcinoma and HPV infection is irrelevant. HPV infection may not be thereason of esophageal squamous cell carcinoma.2p16genes exon2and ESCC have some relationships.3The later pathology stages of ESCC Esophageal, the more p16genes exon2mutations. p16genes exon2and patients smoking history and lymph nodemetastasis have some relationships.4HPV infection is not the reason of p16exon2mutations.