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Expressions of Pim-3 and Bcl-2 in Human Esophageal Squamous Cell Carcinoma and Their Correlative Analysis

Author: ZhaoHui
Tutor: LiHuiXiang；LvXinQuan
School: Zhengzhou University
Course: Pathology and Pathophysiology
Keywords: Esophageal squamous cell carcinoma Pim-3 bcl-2 Semi - quantitative RT-PCR Western blotting In situ hybridization Immunohistochemistry
CLC: R735.1
Type: Master's thesis
Year: 2010
Downloads: 22
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Abstract

Esophageal cancer is a very high degree of malignancy of the digestive tract tumor more common, is one of the most harmful to human health in the world today malignant Current treatment of esophageal cancer has significantly improved compared to the past, with the improvement of medical standards, . Still the best method of treatment of early esophageal cancer is surgical resection; treatment of advanced esophageal status quo is not optimistic, with partial surgical treatment is difficult to achieve satisfactory results, the treatment of esophageal cancer at this stage of the radiotherapy, chemotherapy and medication, but the recurrence rate is still high, patients with five-year survival rate is still very low, it is difficult to meet the ultimate goal of the current medical development. The current rise of targeted gene therapy has become a new type of alternative means to put, not satisfied with the effect of chemotherapy treatment of cancer patients. Thus, from the molecular level to explore the molecular mechanism of the development of esophageal squamous cell carcinoma, and to find effective therapeutic target for inhibition of esophageal squamous cell carcinoma development, clinical treatment of esophageal squamous cell carcinoma, prevention, and patient survival and quality of life improvement extremely important significance. Pim-3 was originally found in the rat pheochromocytoma cell line PC12 cells, was identified as the gene induced depolarization KID-1, KID-1 and expression of serine / threonine kinase activity of the proto-oncogene Pim family having a high degree of homology, thus the name of Pim-3. In recent years, studies have shown that activation of Pim kinases play an important role in the pathogenesis of malignant tumors, a very close relationship with tumor development, and plays an important role in tumor cell apoptosis in tumor cells wither death mechanism may be a proto-oncogene, Pim-3 phosphorylation of Bad protein serine 112, and further lead to bcl-XL and bcl-2 release, prevent apoptosis. So far, the proto-oncogene Pim-3 and suppression role in the development of esophageal squamous cell carcinoma of the relationship between the apoptotic gene bcl-2 both at home and abroad has not been reported, the study by RT-PCR, Western blotting and in situ four different methods of hybridization and immunohistochemistry combined detection of high incidence of esophageal patients with esophageal cancer in Anyang, Henan Province, surgical removal of the cancer tissue oncogene Pim-3 and antiapoptotic gene bcl-2 mRNA and protein expression analysis its relationship with clinicopathological features, to further explore Pim-3 and bcl-2 expression in esophageal squamous cell carcinoma, the study aims to further explore Pim-3 and bcl-2 in esophageal squamous cell carcinoma development in role and looking for the detection of esophageal squamous cell carcinoma of the new sub-targets lay the foundation. Method 1. Semi-quantitative RT-PCR method to detect esophageal squamous cell carcinoma, adjacent atypical hyperplastic tissue and normal esophageal mucosal epithelial tissue Pim-3 and bcl-2 mRNA relative expression levels. 2. Relative expression levels of esophageal squamous cell carcinoma, adjacent atypical hyperplastic tissue and normal esophageal mucosal epithelial tissue Pim-3 and bcl-2 protein was detected by Western blotting. Using in situ hybridization technique detected 45 cases of esophageal squamous cell carcinoma, adjacent atypical hyperplastic tissue in 22 cases of cancer and 45 cases of normal esophageal mucosal epithelial tissue Pim-3 and bcl-2 mRNA expression, and analysis of protein expression related sex. By immunohistochemical methods detected 45 cases of esophageal squamous cell carcinoma, adjacent atypical hyperplastic tissue in 22 cases of cancer and 45 cases of normal esophageal mucosal epithelial tissue Pim-3 and bcl-2 protein expression, and analysis of protein expression related sex. 5. Statistical analysis: SPSS 13.0 software analysis, using χ2 test, t-test and analysis of variance, and correlation analysis. P lt; 0.05 considered statistically significant. 1.Pim-3 semi-quantitative RT-PCR results showed that the 500bp Pim-3 gene successfully amplified in esophageal squamous cell carcinoma, and no corresponding bands in the normal esophageal mucosal epithelial tissue. Esophageal squamous cell carcinoma and adjacent atypical hyperplastic tissue Pim-3 relative expression abundance were 0.809 ± 0.015 and 0.092 ± 0.009, and expression in normal tissue 0 expression among a significant difference (P lt; 0.05). 2. Bcl-2 Semi-quantitative RT-PCR results showed that 595bp of the bcl-2 gene successfully amplified in esophageal squamous cell carcinoma, and no corresponding bands in the normal esophageal mucosal epithelial tissue. Esophageal squamous cell carcinoma and adjacent atypical hyperplastic tissue, bcl-2 relative expression abundance were 0.671 ± 0.011 and 0.102 ± 0.007, and expression in normal tissue is 0, the expression of the three asked a significant difference (P lt; 0.05). 3. Western blotting results show that Pim-3 and bcl-2 protein is highly expressed in esophageal squamous cell carcinoma, while no expression in normal esophageal mucosa. Pim-3 in esophageal squamous cell carcinoma and of bcl-2 protein relative expression levels were: 0.796 ± 0.018 and 0.757 ± 0.014, significantly higher than the cancer next to atypical hyperplasia Pim-3 and the relative expression of bcl-2 protein ( were: 0.299 ± 0.011 and 0.225 ± 0.009), the normal esophageal mucosa relative expression of Pim-3 and bcl-2 expression among significant difference (P lt; 0.05). 4. Situ hybridization showed that, Pim-3 mRNA is mainly localized in the cytoplasm, blue-purple granules. Pim-3 mRNA in esophageal squamous cell carcinoma, high expression of the positive rate was 82.22% (37/45), significantly higher than in adjacent atypical hyperplasia epithelium and normal esophageal epithelium Pim-3 mRNA expression, both positive rate of 22.73% (5/22) and 0.00% (0/45) were statistically significant differences in expression between the three groups (χ 2 = 82.487, P lt; 0.05). 5.Pim-3 protein is localized in the cytoplasm, pale yellow to brown granules. Pim-3 protein is highly expressed in esophageal squamous cell carcinoma, the positive rate of 75.56% (34/45), significantly higher than the adjacent atypical hyperplastic tissue and normal esophageal epithelium Pim-3 protein expression, the positive rates were : 18.18% (4/22) and 0.00% (0/45), the expression differences between the three groups was statistically significant (χ2 = 72.570, P lt; 0.05). 6 In situ hybridization showed, bcl-2 mRNA was mainly localized in the cytoplasm, blue-purple granules. The number of cases to 35 cases, the positive expression rate: 77.78% (35/45), in adjacent atypical hyperplasia epithelium of the seven cases, the mRNA expression of bcl-2 mRNA positive expression in 45 cases of esophageal squamous cell carcinoma The positive rate was 31.82% (7/22), in normal esophageal mucosa has not yet found the expression of bcl-2mRNA. The positive expression rate of Bcl-2 mRNA in esophageal squamous cell carcinoma, adjacent atypical hyperplastic tissue and normal esophageal mucosa is a statistically significant difference (χ 2 = 58.451, P lt; 0.05). 7. Bcl-2 protein expression was localized in the cytoplasm, pale yellow to brown granules. Immunohistochemical detection results showed that the positive expression of bcl-2 protein in 45 cases of esophageal squamous cell carcinoma in the number of cases to 32 cases, the positive expression rate: 71.11% (32/45), and in adjacent atypical hyperplasia epithelium for the five cases, the expression of the positive rate was 22.72% (5/22), in normal esophageal mucosa have not yet found the expression of bcl-2 protein. The positive expression rate of Bcl-2 protein in esophageal squamous cell carcinoma, adjacent atypical hyperplastic tissue and normal esophageal mucosa in a statistically significant difference (χ 2 = 52.724, P lt; 0.05). 8 using the SPSS 13.0 statistical processing software analysis Pim-3 mRNA expression in esophageal squamous cell carcinoma clinical pathological features of the relationship. The results showed that Pim-3 mRNA expression correlated with lymph node metastasis and TNM stage has a significant correlation (P lt; 0.05), but with the histological grade, depth of invasion unrelated (P gt; 0.05). 9.Pim-3 and bcl-2 correlation analysis showed that, in 45 cases of esophageal squamous cell carcinoma, Pim-3 protein expression in 34 cases, bcl-2 protein expression accounted for 30 cases, while Pim-3 protein negative expression in 11 cases, the negative expression of bcl-2 protein accounted for nine cases. Pim-3 and bcl-2 in esophageal squamous cell carcinoma, the expression intensity was positively correlated (γp = 0.664, P = -0.000). Conclusions. Pim-3 and bcl-2 in esophageal squamous cell carcinoma showed a high expression of Pim-3 and bcl-2 expression in normal esophageal epithelium are missing. Esophageal squamous cell carcinoma Pim-3 and bcl-2 expression and lymph node metastasis and TNM stage, but with the histological grade, depth of invasion, suggesting that Pim-3 and bcl-2 in esophageal squamous cell carcinoma play an important role in the occurrence and development. Esophageal squamous cell carcinoma Pim-3, and the intensity of bcl-2 mRNA and protein expression in esophageal squamous cell carcinoma was positively correlated, suggesting that Pim-3 inhibition of cell apoptosis in the tumor may be close to the expression of bcl-2 related.

Expressions of Pim-3 and Bcl-2 in Human Esophageal Squamous Cell Carcinoma and Their Correlative Analysis

Abstract

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