Expression Of Calcium-related Proteins In Prostate Tumors

Prostate gland plays a vital physiological function in male.In the United States and Western European countries,due to the increase in the elderly population,there is high incidence and high mortality rates of prostate tumor with widespread concerns.In China,there is a high incidence of prostate tumor in males with high mortality rate of malignant tumors as well.As China's population is aging,the incidence of prostate tumor is gradually increased.Early clinical symptoms are not obvious,which leads to tumor being found in the late stages,missing the best period of treatment.In addition,there is the lack of biological targets for the early stages of prostate tumor,resulting in misdiagnosis or late-diagnosis and late treatment,lethality gradually increases in the past decades.Prostate tumor has diverse pathological and clinical manifestations;very long latency period,up to more than ten years or even decades;incidence of occult,early symptoms are not obvious;high incidence and mortality rate,the lack of clear preventive interventions and early diagnosis of prostate tumor makes it a great challenge.High-quality early diagnosis of biological markers for prostate tumor prevention and treatment is very important.At present,some literature shows that the development of calcium-associated proteins and prostate tumors has a correlation.How they are correlated is elusive,therefore,the aim is to study the interaction between prostate tumor and the second messenger Ca2+-related proteins.The expression patterns of calcium channel protein,CAMRs,PP2A and PP2B in the RyR complex and integrated protein integrins,extracellular matrix protein FNs,p53 and PTEN in prostate tumors.A combination of the above methods,including tumor cell culture methods,will provide a strong reference for finding early tumor markers and contribute to a more comprehensive approach for early treatment of prostate tumor personalized.In this study,we employed the J-91 transgenic mice and human prostate tumor samples,mouse and human prostate tumor samples were classified into Group I(Gleason 2-4),Group II(Gleason 5-6)and Group III(Gleason 7-10).The histological changes of prostate tumors and human prostate tumor were observed by routine morphological prostate tumors were detected by immunohistochemistry.The expression of CAMRs,PP2A,PP2B family,p53 and PTEN were detected by Western Blot.The expression of CAMRs and integrin aV in human and prostate tumor were detected by immunohistochemical method.By using Real-time quantitative PCR,the mRNA expressions of CAMRs in three levels of prostate tumor were detected.The isolated cells were analyzed with high-content cell imaging analysis system.The results are as follows:(1)Routine histological staining showed that the prostate gland structure of prostate tumor mice and human were abnormal in compared with normal wild type,and the basal cell layer was gradually disintegrated or even disappeared.Collagen fiber deposition showed that the degree of tumor fibrosis deepened.As prostate epithelial cells transformed into tumor cells,the number of nuclei and the volume of the cells.The blood vessels,fat,lymphatic vessels also increased in these prostatic tissues.(2)Immunohistochemical analysis showed that the expression of CAMR1 and CAMR2 protein in the normal prostate tissue was less,while in the tumor cells were more,which mainly located in the basal membrane and epithelium in mice.The CAMRl,CAMR2 and integrin aV was expressed in the basal membrane and epithelium of human tumor cells.The expression of integrin ?V in human tumor cells was higher than that in human primary tumor.(3)The expression of PP2A A,PP2A C,PP2B B,FN-3,and PTEN was significantly up-regulated(P<0.05).Although,the expression of CAMR1 was up-regulated,which was significant(P<0.01).Furthermore,the expression of CAMR2,PP2A B,PP2A B55 and FN-1,FN-4 and FN-5 increased,whereas,the expression of PP2B A and p53 decreased in tumor cells,which were not significant.(4)Real-time quantitative PCR results showed that CAMR1 and CAMR2 were expressed higher in the three-stage tumor than in normal tissues,which was not significant.(5)The results of high content test showed that the parameters such as the speed and distance of cell movements,size in area of the cells,and average fluorescence intensity of prostate tumor cells were significantly higher than control cells on both the ECM and ordinary dishes.Statistical analysis showed that tissue,dispersed and gathering cells were isolated from G?,G? and G? tumor-derived cells in both cell area and length,cell movement distance and velocity,the nucleus area and the average fluorescence intensity of the high content parameters displayed a very significant difference both in the ECM and ordinary dishes(P<0.001).Conclusions:Prostate tumor is a malignancy of the male reproductive system,and lack of early biomarkers leads to high morbidity and mortality.Studies have reported that calcium-associated proteins is related with the development of prostate tumor,and calcium ions in the second messenger play an important role in cell division,metastasis and apoptosis.In this study,the expression patterns of Ca2+-associated protein,extracellular matrix protein FNs,PTEN and p53 in prostate tumor were investigated by immunohistochemistry,real-time quantitative PCR and Western Blot.The results suggest that calcium-associated proteins are associated with the occurrence of prostate tumor and are likely to be potential biomarkers.A combination of the above methods,including tumor cell culture methods,will help in the preliminary diagnosis of prostate tumor.This research will provide the experimental and theoretical basis for future works.