The Expression Of Ki67、CD133、Ezrin And RhoA In SW-13 Cell Line And Adrenal Cortex Tumor And Its Clinical Significance

OBJECTIVE To investigate the expression of proliferating cell nuclear antigen Ki67、cancer stem cell marker CD 133、invasion and metastasis associated proteins Ezrin and RhoA in SW-13 cell line in vivo and in vitro and adrenal cortex tumors, analyzing their relationship with clinical pathological parameters. For studying the biological significance of these factors in the tumorigenesis and development of adrenal cortex tumor.METHODS1、Basic research:to establish subcutaneous tumor model and peritoneal metastasis mode, ten nude mice were inoculated subcutaneously in both sides of inguinal region and the other ten nude mice were inoculated intraperitoneally with 1×107/ml SW-13 cells. Analyzing and comparing the characteristics of two models. Histopathological features were examined by HE staining, and the expression of Ki67、CD133、Ezrin and RhoA were detected by immunohistochemistry in SW-13 cells、transplanted tumors and metastatic tumors.2、Clinical pathological specimens research:Ki67、CD133、 Ezrin and RhoA were detected by immunohistochemistry in 64 cases of adrenal cortical tumors (9 cases of adrenocortical hyperplasia,39 cases of ACA,16 cases of ACC) and 10 cases of normal adrenal cortex tissue, to analyze their relationship with clinical pathological parameters and their expression correlation.RESULTS1、It was successful to construct subcutaneous tumor model and peritoneal metastasis model. Subcutaneous tumors were found in both sides of inguinal region of ten nude mice, the tumor-take rate was 100%. Bilateral adrenal glands of ten nude mice increased in size when dissected. Peritoneal metastasis appeared in the nude mouse whose subcutaneous tumor was the biggest. Neoplasias were found in local inoculation of abdominal walls of ten nude mice, the tumor-take rate was 100%. Peritoneal metastatic tumor nodules mainly disseminated to mesentery and greater omentum. Pancreatic metastases were discovered in three mice and renal metastases in two. By HE staining, pathological mitotic figures were common, and neoplastic multinucleated giant cell was found occasionally. Metastatic tumor nodules were identified as mesenteric lymph nodes metastases, the complete structures had been destroyed. Pancreas and kidney were infiltrated by the cancer cells. The bilateral adrenal sizes were reactive increase.2、Ki67、Ezrin and RhoA strong positively or positively expressed in SW-13 cell line in vivo and in vitro. CD 133 positively expressed in SW-13 cell line in vitro, and weakly expressed in transplanted tumor and metastatic tumor.3、Normal adrenal cortex tissue and adrenocortical hyperplasia were not detected positive expression of Ki67. The positive rate of Ki67 in ACA and ACC were 12.8%(5/39)、68.8%(11/16).The positive expression level of Ki67 in ACC was significantly higher than ACA、hyperplasia group and nomal group (all P=0.000). The positive expression of Ki67 did not correlate with gender、 age、blood pressure、capsular invasion and distant metastasis(P>0.05), but was closely related with tumor size、mitotic rate and necrosis (P<0.01); The positive rate of CD 133 in normal adrenal cortex tissue、adrenocortical hyperplasia、ACA and ACC were 30%(3/7)、33.3%　(3/9)、35.9%(14/39)、 37.5%(6/16), and there was no significant difference in four groups (P>0.05). The positive expression of CD 133 did not correlate with gender、age、blood pressure、tumor size、capsular invasion、mitotic rate、necrosis and distant metastasis(P>0.05); The positive rate of Ezrin in normal adrenal cortex tissue、 adrenocortical hyperplasia and ACA were 20%(2/10)、22.2%(2/9)、43.6% (17/39), it mainly distributed on cell membrane. The positive rate of Ezrin in ACC was 75%　(12/16), it distributed on both cell membrane and cytoplasm. The positive expression level of Ezrin in ACC was significantly higher than ACA。 hyperplasia group and nomal group (P<0.01).The positive expression of Ezrin did not correlate with gender、age、blood pressure、tmitotic rate、necrosis and distant metastasis (P>0.05), but was closely related with tumor size and capsular invasion (P<0.05); Normal adrenal cortex tissue were not detected positive expression of RhoA, The positive rate of RhoA in adrenocortical hyperplasia、ACA and ACC were 11.1%(1/9)、33.3%　(13/39)、56.3% (9/16).The positive expression level of RhoA in ACC was significantly higher than ACA、hyperplasia group and nomal group(all P<0.01), The level of RhoA expression in ACA was higher than nomal group (P<0.05). The positive expression of RhoA did not correlate with gender、age、blood pressure、mitotic rate and necrosis(all P>0.05), but was closely related with tumor size、capsular invasion and distant metastasis (P<0.05)4、In adrenal cortex tumor, Ki67 expression level was positively correlated with Ezrin、RhoA (γ=0.762、0.661, P<0.01), there also was positive correlation between Ezrin and RhoA (γ=0.745, P<0.01)5、Evaluation of diagnostic test:the combined sensitivity of Ki67、Ezrin and RhoA was 81.3%, the combined specificity was 89.6%, the combined positive likelihood ratio was 7.82, the combined negative likelihood ratio was 0.19, the combined youden’s index was 0.709.CONCLUSION1、Subcutaneous tumor model and peritoneal metastasis model in nude mice were established by SW-13 cell line, which provided meaningful animal models for studying the affection of different tissue microenvironment on the expression of Ki67、CD133、Ezrin and RhoA in SW-13 cells.2、 Ki67、Ezrin and RhoA highly expressed in ACC, which could serve as the important references for evaluating the malignant potential of adrenal cortex tumor. The accuracy of distinguishing the malignant from benign adrenal cortex tumor was enhanced by combined detection of these three indicators.