| Objective: Esophageal cancer(EC) is the sixth leading cause of cancerrelated death globally. In high-risk areas such as Henan and Hebei, esophageal squamous cell carcinoma(ESCC) accounts for 90% of esophageal cancers. Malnutrition, low intake of fruits and vegetables, drinking hot beverages, and smoking all contribute to the etiology of ESCC. Human papillomaviruses(HPV) are small, double stranded DNA viruses that infect epithelial tissues, including those of the oral,esophageal and anogenital tract. Nowadays some studies have shown that HPV infection is one of risk factors in the development of ESCC. It has been proved that HPV early protein can inhibit the apoptosis pathway of Fas/Fas L and further cause the malignant transformation in cervical carcinoma. But the role of HPV infection in esophageal cancer remains controversial. Therefore, to make sure the role of HPV infection in esophageal cancer, in this study, we analyzed the infection rate of HPV and evaluated the expression of Fas and Fas L in ESCCs. We also investigated the Relationship between infection of HPV and espression of Fas and Fas L on ESCC.Method:1 CasesThis study included 150 patients with ESCC, which were consecutive cases seen at NO.2 Hospital of Hebei Medical University, China between 2009.01 and 2014.05 who had histopathologically confirmed primary ESCC. Patients who had primary tumors outside of the esophagus, tumors of unknown origin, or any histopathologic diagnosis other than ESCC were excluded. Clinicopathological Features of 150 ESCCs include: age, gender, lymph node metastasis and histological differentiation.2 DNA isolation and direct sequencingDNA was extracted from formalin fixed paraffin embedded(FFPE) samples. HPV was analysed by PCR, which was used for the amplification of the HPV L1(MY09/MY11) gene in all samples, with proper negative and positive PCR controls in both amplifications. PCR was used to amplify the housekeeping gene b-actin(200 bp) to assess the presence of PCR inhibitors in the samples used in this study. HPV positive controls were obtained from cervix tumor FFPE samples, which had been ensured HPV infection. HPV negative controls were dd H2 O instead of HPV L1 primers in PCR.3 Detection the expression of Fas/Fas L by immunohistochemistryThe expression of Fas and Fas L protein was investigated by SP immunohistochemical staining method. Fas and Fas L protein positive reaction mainly located in cell membrane, a small amount in the cytoplasm.4 StatisticsThe experimental data were analyzed with Chi-square test correlation with stastics software of SPSS19.0 edition.Results:1 150 ESCC DNA samples were analyzed for the presence of HPV by PCR, with 51 of them presenting a positive amplicon with 34.0% being positive.2 Expression of Fas/ Fas L in different groups: The positive expression rate of Fas was 59.3% in ESCC lower than 94.7% in adjacent normal oesophageal epithelium(P<0.05).The positive expression rate of Fas L was 76.0% in ESCC higher than 44.7% in adjacent normal oesophageal epithelium(P<0.05).3 Expression of Fas/Fas L in HPV infected tissues: The positive expression rate of Fas in ESCC with HPV infected tissues was 25.5% lower than 76.8% in HPV uninfected tissues. The positive expression rate of Fas in adjacent normal oesophageal epithelium with HPV infected tissues was 86.3% lower than 99.0% in HPV uninfected tissues(P<0.05). The positive expression rate of Fas L in ESCC with HPV infected tissues was 56.9% lower than 85.9% in HPV uninfected tissues. No difference in the Fas L expression was found in adjacent normal oesophageal epithelium between HPV infected tissues and HPV uninfected tissues(P>0.05).4 Relationship beween HPV infection and clinicopathological features: in The poorly differentiated rate of ESCC with HPV infection was 43.1% higher than 22.2% in the poorly differentiated rate of ESCC with HPV uninfection(P<0.05). The well/ moderate differentiated rate of ESCC with HPV infection was 56.9% lower than 77.8% in the well/moderate differentiated rate of ESCC with HPV uninfection(P<0.05). No difference in relationship of HPV infective among age, gender and lymph node metastasis(P>0.05).5 Relationship beween expression of Fas/Fas L and clinicopathological Features: The expression of Fas in poorly differentiation of ESCC was 43.2% lower than 66.0% in well/moderate differentiation(P<0.05). No difference in relationship of expression of Fas among age, gender and lymph node metastasis(P>0.05). The expression of Fas L in poorly differentiation of ESCC was 61.4% lower than 82.1% in well/moderate differentiation(P<0.05), and the positive expression rate of Fas L was 65.8% in more than 62 years old(include 62) lower than 87.3% in less than 62 years old(P<0.05). No difference in relationship of expression of Fas L beween gender and lymph node metastasis(P>0.05).6 Relationship among HPV infection, expression of Fas/Fas L and clinicopathological features: The positive expression rate of Fas in ESCC with HPV infection was 13.8% in more than 62 years old(include 62) lower than 40.9% in less than 62 years old(P<0.05). The positive expression rate of Fas L in ESCC with HPV infection was 41.4% in more than 62 years old(include 62) lower than 77.3% in less than 62 years old(P<0.05).Conclusions:1 HPV infective rate is 34.0% in ESCC. There are more poorly differentiation in ESCC with HPV infection. That means HPV was correlated with tumor differentiation.2 In ESCC, the expression of Fas is decreased, but the expression of Fas L is increased. That means the abnormal expression of Fas/Fas L was correlated with the occurrence of ESCC.3 The expression of Fas/Fas L in ESCC with HPV infectived were all decreased, which means HPV was correlated with the occurrence of ESCC though inhibited cell apoptpsis.4 The expression of Fas/Fas L in aged patients with HPV infection is lower, which means the influence of HPV infection in different populations were not exactly the same. There may be a sensitive crowd. |
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