| Background:The endometrial adenocarcinoma is one of the three great malignant tumours in woman reproductive tube. In recent years,there is the trend of escalating incidence in the world wide. About producing mechamisms of endometrial adenocarcinoma are not yet very clear,thus exploring serum biomarkers and searching treatment molecular way become very focus. These studies will be very important to the early clinical diagnosis, treatment and prognosis evaluation of the endometrial cancer. Moreover,it could provide evidence to further explore the pathogenesis of endometrial cancer.Objective:To investigate and compare the expression of sperm-associated antigen 9 (SPAG9) in normal endometrial tissues,endometrial hyperplasia,endometrial carcinoma and adjacent tissues. To detect the humoral immune response of sperm-associated antigen 9 (SPAG9) in human serum. Evaluating its potential value as a serum biomarker. Discussion the possible relationship and its significance between SPAG9 and endometrial cancer.Methods:1. Subjects:Collecting the endometrial tissues of surgical patients,including 47 cases of endometrial cancer (contain 12 cases of adjacent tissues); 10 cases of atypical endometrial hyperplasia;12 cases of endometrial hyperplasia;20 cases of normal endometrium.Collecting the serum of 50 endometrial cancer patients which have 47 cases are corresponding with the cases of tissue samples.Control group serum samples were collected from 10 healthy women,20 benign tumor patients(8 uterine fibroids,7 ovarian cysts, and 5 endometrial polyps).2. Test:(1) Immunohistochemistry (IHC) was used to detect the expression of SPAG9 protein in normal endometrium, endometrial hyperplasia, endometrial cancer tissue and adjacent tissues. To clarify its localization in tissues.(2) Reverse Transcription-Polymerase-Chain Reaction(RT-PCR) was used to detect the expression of SPAG9 mRNA in normal endometrium, endometrial hyperplasia, endometrial cancer tissue and adjacent tissues, and using semi-quantitative analysis.(3) Western blot was used to detect the expression of SPAG9 protein in normal endometrium, endometrial hyperplasia, endometrial cancer tissue and adjacent tissues, and comparing the differences use semi-quantitative analysis.(4) SPAG9 concentration in serum samples from 10 healthy women,10 women with benign diseases and 40 women with endometrial carcinoma was detected by ELISA.Results:1. Immunohistochemistry revealed that SPAG9 protein was localized in the cytoplasm of endometrial cancer, the interstitial tissue has no or little expression. SPAG9 protein expression was observed in 72% tissues of endometrial cancer, which is significantly high than in atypical hyperplasia tissues (30%)(χ2=4.764, P=0.029< 0.05) and endometrial hyperplasia tissues(25%)(χ2=7.249, P=0.007<0.05). The adjacent tissues and normal tissues have no expression. It was also expressed in transferred lymph nodes.2. RT-PCR results showed SPAG9 mRNA in endometrial carcinoma with high expression (0.833±0.667), and SPAG9 gene in tumors with high expression in early stage;The average expression level was significantly higher than other tissues (P <0.05); The adjacent tissues and normal tissues have no expression. The expression of SPAG9 was low in atypical hyperplasia (0.089±0.026) and simple hyperplasia (0.009±0.001), they were all had significant difference compared with cancer tissues(P< 0.05), however, it has no significant difference (P>0.05) between them.3. Western blot results were consistent with immunohistochemistry and RT-PCR results. SPAG9 protein in endometrial carcinoma with high expression (0.656±0.697), the expression levels of adenocarcinoma(0.797±0.762) were significantly higher when compared with serous papillary carcinoma(0.219±0.183)(P<0.05);The adjacent tissues and normal tissues have no expression; The expression of SPAG9 was low in atypical hyperplasia (0.232±0.201) and simple hyperplasia (0.032±0.006), it has no significant difference (P> 0.05) between them,but they were all had significant difference compared with cancer tissues(P<0.05).4. SPAG9 serum concentration (ng/ml) (mean±standard deviation) in normal healthy women is 5.4256±1.52587, in patients with benign tumors was 6.328±1.28181, SPAG9 serum concentration in the serum of patients with endometrial cancer was 19.0114±13.19338, which was significantly higher than the former two (P<0.01).Conclusion:1. SPAG9 protein and mRNA in tumors with high expression in early stage,its expression levels were significantly higher in endometrial carcinoma when compared with atypical endometrial hyperplasia and endometrial hyperplasis; the adjacent tissues and normal tissues have no expression.The expression between atypical endometrial hyperplasia and simple hyperplasia showed no significant differences2. The positive expression of SPAG9 was associated with pathological, and it was also expressed in transferred lymph nodes.3. SPAG9 serum concentration in patients with endometrial cancer was significantly higher than in normal healthy women and patients with benign tumors.4. SPAG9 may serve as a new type of endometrial cancer markers for early detection, diagnosis and prognosis evaluation.a... |
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