| Objective:1. To identified VM structure existence in human orbital adenoid cystic carcinoma tissue samples and investigate the correlations with pathological type and meanings.2. To observe the expression and distribution of CD44v6, CD133, and ABCG2 in orbital ACC and investigate their correlations with pathological type and VM, which can explain the trend to recur and transfer in local.3. Carried on the morphology observation in salivary adenoid cystic carcinoma cell line SACC-83, to make sure that VM exists in SACC-83 and to observe the expression and distribution of CD44v6, CD133, and ABCG2 in SACC-83, which will build foundation for the follow-up studies to get TSCs in ACC and analyse the forming mechanism of VM.Method:1.This experiment accumulates thirty three ACC cases which undergown surgery in the 2nd affiliated hospital of Tianjin Medical University from January 1999 to March 2009. All samples were sectioned and stained by HE to confirm the pathology of the tumors. Immunohistochemical staining, PAS-CD31 double staining were performed to observe the structure and distribution of VM..At the same time, two stages method of immunohistochemical staining was employed in 33 cases of human orbital ACC, to observe the expression and distribution of CD44v6, CD133.and ABCG2.2.According to the observation in the lachrymal ACC,. we found VM in different pathological type and used statistical methods to compare the correlation between VM and CD44v6. CD133,ABCG2.3.In two-dimensional cell culture and three-dimensional cell culture of SACC-83. we observe the expression and distribution of VM and CD44v6, CD133, ABCG2 in salivary adenoid cystic carcinoma cell line SACC-83.Results:1. All the cases were diagnosed as orbital ACC by HE staining and immunohistochemical staining results.33 cases included 13 solid type (39.39%).20 adeno-tubiform type (60.61%).10 cases appeared VM structure, the rate is 30.3%. solid type 6 cases (46.2%), adeno-tubiform type 4 cases(20.0%), There wasn't statistically different between them (p=0.346).2 The VM is formed by ACC cells as tubular structure, which contains RBC. The red blood cells have complete structure, around which exist no bleeding areas. In addition, a part of tumor cells'cytoplasm endocrining CD31 and PAS material were observed.3. The positive rate of CD44v6 was 54.5% (18/33), with 76.9%(10/13) in solid type and 40.0%(8/20) in adeno-tubiform type. There wasn't statistically different between them (p=0.037). The positive rate of CD133 was 57.6%(19/33), with 76.9%(10/13) in solid type and 45% (9/20) in adeno-tubiform type. There was statistically different between them (p=0.070). Parts of the expression of CD133 antigen were both on the cytoplasm and the nucleus. The positive rate of ABCG2 was 21.2% (7/33), with 30.8% (4/13) in solid type and 15% (3/20) in adeno-tubiform type. There wasn't statistically different between them (p=0.297). Lots of positive cells surrounded vessels in tumor tissues.4. In two-dimensional cell culture of SACC-83, the SACC cells grew like loops and nets. In three-dimensional cell culture of SACC-83, the SACC cells grew like tubes and cavities. VM were found in three-dimensional cell culture of SACC-83.The positive rates of CD44v6, CD133, ABCG2 in SACC-83. separately are 42.47±7.37%,56.56±10.27% and 2.94±1.23%Conclusion:1. VM structure exists in both human orbital ACC and three-dimensional cell culture of SACC-83;2. In human orbital ACC, the different pathological types did no directted impact on VM, but related with CD44v6;3. In human orbital ACC, the results showed that rate of VM appeared in ACC is related with CD44v6.that means VM is not only apt to recur and transfer in local.but also has close relations with CD44v6.4. The expression of CD44v6, CD133 and ABCG2 in ACC should have some significance in the progress of ACC. And it could be regarded as one of the factors to find TSCs in ACC;... |
