Expressions Of Pokemon, MDM2 And P53 Protein In Esophageal Squanous Cell Carcinoma And Their Clinical Significance

Objective: China is a country that has high incidence and mortality of esophageal cancer in the world.For many years,the high incidence and mortality of esophageal cancer seriously has threatened national life and health.Different from the European and American countries,esophageal cancer pathological histology of Chinese residents is mainly for squamous cell carcinoma which accounted for more than 90 percent.Currently accepted,early detection,early diagnosis and early treatment is the most effective measures for prevention and control of esophageal cancer,and the early detection and early diagnosis is the key to realize the early treatment.Right now,a single tumor marker has limited to the early diagnostic value of esophageal cancer in clinic,then more research is focused on the multiple tumor markers combined detection to improve the early diagnostic rate.Therefore,our study choosed esophageal squamous cell carcinoma(ESCC)as the research object and Pokemon,MDM2 and p53 gene in the Pokemon→ ARF→ MDM2→ wtp53(Wild type p53)Cancer pathways as the indicators,to detecte the expressions of Pokemon,MDM2 and p53(including Wild type p53 and Mutant type p53)protein in ESCC tissus and normal esophageal mucosa tissus using Immunohistochemical Streptomyces avidin peroxidase connection method,and analyzed the relationship between Pokemon,MDM2 and p53 protein and the clinical pathological features of Patients with esophageal cancer.Methods:1 Esophageal squamous carcinoma tissues and normal esophageal mucosa tissues were derived from the 62 cases of ESCC patients in the First Hospital of Hebei Medical University,who underwent surgery surgical resection but not received preoperative chemotherapy,radiation therapy,and other associated with anti-tumor treatment during January in 2010 to December in 2014.39 cases were men,23 cases were women,aged 48 to 75 years old,mean age was 63.13±5.67 years old.All subjects were confirmed by two independent chief physicians in pathology department.The subjects with consistent diagnosis were included in the study.Groups: In the 62 cases of ESCC,10 cases were highly differentiated squamous carcinoma,28 cases were moderately differentiated squamous cell carcinomas,24 cases were low differentiated squamous cell carcinoma and 35 cases of adjacent normal tissues as the normal control group.Above them,36 cases with fibrosa infiltration,26 cases with no fibrosa infiltration;24 cases with lymph node metastasis,38 cases with no lymph node metastasis;42 cases with Ⅰ~Ⅱ stage in TNM stage,20 cases with Ⅲ~Ⅳ stage.2 Routine histological processing: all specimens were fixed in 10% formalin solution,embedded in paraffin and sliced,HE staining was for pathological examination.Histological grade was according to TNM staging system of ESCC formulated by the American Joint Committee on Cancer(AJCC)/ International Union Against Cancer(UICC).3 Immunohistochemistry: Tissues were paraffin-embedded and sliced continuously with 4um thickness,oven 60℃~65 overnight,dewaxed,citric ℃repair liquid boiling to repair,used SP immunohistochemical staining according to immunohistochemistry kit manual steps strictly,colored with DAB,hematoxylin staining,known Pokemon protein,MDM2 protein,and p53 protein positive slice as positive control,PBS was used instead of primary antibody as a negative control.4 Interpretation of the results: used double-blind method to observe results under a microscope.Pokemon and MDM2 and p53 protein is nucleus staining.Positive determination was judged by positive cells and staining intensity.5 Statistical treatment: The experiment data were analyzed by using SPSS 16.0 software.The χ2 test was applied in the intergroup comparison of enumeration data.Correlation detection was used in Spearman rank correlation analysis.P<0.05 was considered that difference was statistically significant.Results: 1 The relationship between the expression of Pokemon protein in two groups and the clinical pathological features.1.1 Pokemon protein comparison between the groups: Pokemon protein expression was 67.74%(42/62)in ESCC,the paracarcinoma normal control group was 42.86%(15/35).The expression of Pokemon protein in ESCC was significantly higher than paracarcinoma normal control group,the difference was statistically significant(P<0.05).1.2 Relations of Pokemon expression in esophageal squamous cell carcinoma and its clinicopathological features: The expression of Pokemon in esophageal squamous cell carcinoma was related with patients with esophageal squanous carcinoma differentiation degree,lymph node metastasis and TNM staging(P<0.05),its expression in the patients with low differentiated squamous cell carcinoma were significantly higher than those in patients with highly differentiated squamous carcinoma,in patients with lymph node metastasis were significantly higher than in patients without lymph node metastasis,in patients with clinical stage III+IV were significantly higher than I+II patients,but no relation with sex,age,fibrosa infiltration(P>0.05).2 The relationship between the expression of MDM2 protein in two groups and the clinical pathological features.2.1 MDM2 protein comparison between the groups: MDM2 protein expression was 54.84%(34/62)in ESCC,the paracarcinoma normal control group was 28.57%(10/35).The expression of MDM2 protein in ESCC was significantly higher than paracarcinoma normal control group,the difference was statistically significant(P<0.05).2.2 Relations of MDM2 expression in esophageal squamous cell carcinoma and its clinicopathological features: The expression of MDM2 in esophageal squamous cell carcinoma was related with patients with esophageal squanous carcinoma differentiation degree,lymph node metastasis and TNM staging(P<0.05),its expression in the patients with low differentiated squamous cell carcinoma were significantly higher than those in patients with highly differentiated squamous carcinoma,in patients with lymph node metastasis were significantly higher than in patients without lymph node metastasis,in patients with clinical stage III+IV were significantly higher than I+II patients,but no relation with sex,age,fibrosa infiltration(P>0.05).3 The relationship between the expression of p53 protein between groups and the clinical pathological features.3.1 p53 protein comparison between the groups: p53 protein expression was 72.58%(45/62)in ESCC,the paracarcinoma normal control group was 17.14%(6/35).The expression of p53 protein in ESCC was significantly higher than paracarcinoma normal control group,the difference was statistically significant(P<0.05).3.2 Relations of p53 expression in esophageal squamous cell carcinoma and its clinicopathological features: the expression of p53 in esophageal squamous cell carcinoma was related with patients with esophageal squanous carcinoma differentiation degree,fibrosa infiltration,lymph node metastasis and TNM staging(P<0.05),its expression in the patients with low differentiated squamous cell carcinoma were significantly higher than in patients with highly differentiated squamous carcinoma,in patients with fibrosa infiltration were significantly higher than in patients without fibrosa infiltration,in patients with lymph node metastasis were significantly higher than in patients without lymph node metastasis,in patients with clinical stage III+IV were significantly higher than I + II patients,but no relation with sex,age(P>0.05).4 The correlation among Pokemon,MDM2 and p53 protein.Spearman rank correlation analysis shows that the expressions of Pokemon and MDM2 protein in ESCC present positive correlation(r=0.599,P<0.01).Pokemon and p53 protein present positive correlation(r=0.501,P<0.01).MDM2 and p53 protein present positive correlation(r=0.359,P<0.01).Conclusions:1 The positive expression rate of Pokemon,MDM2 and mtp53(Mutant type p53)protein in ESCC were significantly higher than paracarcinoma of normal esophageal mucosa group,suggesting that Pokemon,MDM2 and mtp53 protein play a role of carcinogenesis in the occurrence of ESCC,and have the potential to be of tumor markers early diagnosis.2 The positive expression rate of Pokemon and MDM2 protein were related to esophageal squanous carcinoma differentiation degree,lymphatic metastasis and TNM stage.The positive expression rate of mtp53 protein was related to esophageal squanous carcinoma differentiation degree,whether the fibrosa infiltration was invaded,lymphatic metastasis and TNM stage,suggesting that there is a certain value of Pokemon,MDM2 and mtp53 protein to judge malignant degree and prognosis of esophageal squamous carcinoma.3 The expressions of Pokemon,MDM2 and mtp53 protein present positive correlation with each other,suggesting that the three proteins play an synergistic carcinogenic roles in the process of esophageal squamous carcinoma pathogenesis.