Expression Patterns Of CAMR1 And CAMR2 In Mouse Soft Tissue Sarcoma

Soft tissue sarcomas are a heterogeneous group of malignant tumors which arise predominantly from the embryonic mesoderm. Soft tissue sarcomas can occur anywhere in the body and have many pathological types and diverse clinical presentation. The molecular mechanism of soft tissue sarcomas has not been identified. All of those features led to the diagnosis of soft tissue sarcomas are more difficult. Therefor, new biological markers are necessary to been established, which are simply operation, specificity and sensitivity. CAMR family is a group of highly homologous receptor binding proteins, belong to the immunophilin super-family. These proteins play important roles in various physiological processes, such as cell cycle, survival and apoptotic signaling pathways, particularly in cancer. The deregulation of their expression has been observed in cancer, such as breast cancer, prostate cancer, pancreatic cancer and colorectal cancer. A wealth of data in vitro and clinical studies is paving the approach for novel, promising roles of CAMRs in diagnostic, prognostic or therapy-monitoring cancer. CAMR1 and CAMR2 are two main members of CAMRs family. The molecular weights were respectively 12kDa and 12.6kDa. At present, there are many researches about the expression of CAMR 1 and CAMR2 in tumpr tissues, but the research about CAMR1 and CAMR2 on soft tissue sarcomas has not been reported.In our study, we selected the transgenic mice whose background is BALB/C as aninmal model. The normal hypodermis and soft tissue sarcomas distributed in mice ear and neck were collected in different stages. According to haematoxylin-eosin (HE) staining^ periodic acid-silve metheramin (PASM) staining and Trichrome staining, the pathological changes of soft tissue sarcoma in mice ear and neck were observed. The expression and localization of CAMR1 and CAMR2 in mice ear and neck soft tissue sarcomas were compared by RT-PCR, Western Blot and immunohistochemistry. The expression levels of CAMR1 and CAMR2 in transgene mice tail blood and liquid of different periods in mice ear and neck soft tissue sarcomas were compared by Western Blot We investegated the localization and expression level of CAMR1 and CAMR2 in mice soft tissue sarcomas, in order to find the changes of expression pattern of CAMR1 and CAMR2 in the process of mice soft tissue sarcomas, to provide quick and effective biological marker for the diagnosis of soft tissue sarcomas. The main research results are as follows:1. RT-PCR showed that in mRNA levels CAMR1 and CAMR2 were transcripted during every stage of mice ear and neck soft tissue sarcomas within the tumor deveploment. This result indicated that CAMR1 and CAMR2 may play same or similar function in mice ear and neck soft tissue sarcomas2. The expression patterns of CAMR1 and CAMR2 proteins were compared by Western Blot analysis:(1) CAMR1 and CAMR2 were detected in every stage of mice ear and neck soft tissue sarcomas. And compared with normal hypodermis, the expression amount of CAMR1 and CAMR2 were down-regulated in soft tissue sarcomas.(2) CAMR1 and CAMR2 were detected in every stage of transgene mice tail blood. With the appearance of tumor in transgenic mice, the expression amount of CAMR1 and CAMR2 were up-regulated, and it reached the maximum in middle transgene mice tail blood and started to decrease in middle-advanced and advanced transgene mice tail blood, but it was always higher than the normal mice tail blood.(3) CAMR1 and CAMR2 were detected in every stages of liquid in mice ear and neck soft tissue sarcomas. The expression amount increased from early stage to advanced stage.3. Histological analysis:HE staining showed that mice soft tissue sarcomas have significant pathological changes in morphology. They were anomalous cellular morphology and distribution, obvious morphological differences in different stages, increased interstitial vascular, easy haemorrhage and necrosis. PASM staining indicated that the broken or missing phenomenon occurred in basement membrane of soft tissue sarcomas, and the degree of fibrosis increased in soft tissue sarcomas. Trichrome staining further demonstrated that the distribution of collagen fibers expanded and the degree of fibrosis increased.4. Immunohistochemical analysis showed that CAMR1 and CAMR2 protein mainly located in the vascular in mice normal hypodermi. In the mice ear and neck soft tissue sarcomas, they were mainly located in the vascular and the cytoplasm of some tumor cells.In conclusion:according to the data mensured with RT-PCR, Western Blot and Immunohistochemical, we identified that CAMR1 and CAMR2 expressed in every stage of mice ear and neck soft tissue sarcomas and the expression amount were lower than normal hypodermis. They mainly located in the vascular and the cytoplasm of some tumor cells. We also detected that CAMR1 and CAMR2 expressed in every stage of transgene mice tail blood and liquid in mice ear and neck soft tissue sarcomas by Western Blot. The expression amount of CAMR1 and CAMR2 in transgene mice tail blood was higher than normal mice tail blood. And the expression amount of CAMR1 and CAMR2 in liquid increased form early stage to advanced stage. All of those showed that CAMR1 and CAMR2 may play important roles in the occurrence and development of soft tissue sarcomas. They and their chelate may become the effective and fast biomarkers for the diagnosis and treatment of soft tissue sarcomas. Further research is necessary to clarify the mechanism of CAMR1 and CAMR2 in soft tissue sarcomas.