ObjectiveTo observe the possibility and mechanism of microRNA (miRNA)-203 inducing the human epidermal stem cells to differentiate into sweat gland cells. MethodsFive normal human foreskin tissues were harvested to prepare a single cell suspension by 0.25% trypsin-EDTA digestion method, then the human epidermal stem cells were isolated and cultured by type IV collagen differential adherent method. The cell morphology was observed by inverted phase contrast microscope. The monoclonal antibodies of integrin β1 (ITGB1), cytokeratin19 (CK19), CK1, CK10, CK18, and carcinoembryonic antigen (CEA) were used for identification by immunocytochemical staining. Double stranded mimics of has-miR-203 were transfected into the human epidermal stem cells with Lipofectamine 2000 (experimental group) and the human epidermal stem cells transfected with nonsense miRNA mimics served as control group. The monoclonal antibodies of ITGB1, CK19, CK1, CK10, CK18, and CEA were used for identifying the cells after transfection by immunocytochemical staining; the mRNA relative expressions of miRNA-203, P63, ITGB1, CK19, CK1, CK10, CK18, and CEA were detected by real-time fluorescence quantitative RT-PCR before transfected and at 72 hours after transfected. The protein relative expressions of P63, ITGB1, CK19, CK1, CK10, CK18, and CEA were detected by Western blot. The mRNA expression of miRNA-203 and the mRNA and protein expressions of P63 were analyzed respectively with Pearson correlation. ResultsThe CK19 and ITGB1 were positively expressed before transfection, but CK1, CK10, CK18, and CEA were expressed positively after transfection. The mRNA relative expression of miRNA-203 after transfection in experimental group was significantly higher than that before transfection (P<0.05). The mRNA and protein relative expressions of CK1, CK10, CK18, and CEA after transfection in experimental group were significantly higher than those before transfection and control group (P<0.05), while the mRNA and protein relative expressions of P63, CK19, and ITGB1 were significantly lower than those before transfection and control group (P<0.05). These indicators showed no significant difference between the control group and before transfection (P>0.05). The expression level of miRNA-203 was negatively correlated with the mRNA and protein relative expressions of P63 before and after transfection, the correlation coefficients before transfection were -0.91 (t=3.862, P=0.042) and -0.96 (t=5.971, P=0.009) respectively; the correlation coefficients after transfection were -0.92 (t=4.283, P=0.031) and -0.95 (t=5.842, P=0.011) respectively. ConclusionmiRNA-203 can induce epidermal stem cells to differentiate into sweat gland cells by targeting inhibition of P63 probably.
【摘要】 目的 研究ΔNP63/TAP63在上皮性卵巢腫瘤組織中的表達及其與臨床病理特征的關系。 方法 運用熒光定量聚合酶鏈反應方法檢測2002年-2004年54例卵巢上皮性腫瘤中ΔNP63/TAP63的基因水平。 結果 33例卵巢上皮細胞癌組織中ΔNP63的表達高于21例良性上皮性腫瘤中組織。卵巢上皮細胞癌中的表達強度與腫瘤組織病理學分期相關（Plt;0.05），良性腫瘤的表達低于惡性腫瘤（Plt;0.05）。ΔNP63的表達高于TAP63（Plt;0.05）；各組間TAP63的表達差異無統計學意義（Pgt;0.05）。 結論 ΔNP63在上皮性卵巢癌中高表達，可能成為上皮性卵巢癌診斷及預后的分子標志物。【Abstract】 Objective To explore the expression of ΔNP63 / TAP63 in human epithelial ovarian tumor tissues and its relationship with the clinicopathological features. Methods Fluorescent quantitative PCR method was used to detect 54 cases of ΔNP63 / TAP63 gene level in 54 patients with epithelial ovarian tumors diagnosed between 2002 and 2004. Results ΔNP63 expression in the 33 cases of ovarian epithelial cell carcinoma was higher than that in the 21 cases of benign epithelial tumor tissue. The expression in ovarian epithelial cell carcinoma was concerned with the pathological staging of tumor (Plt;0.05); the expression in benign tumors was lower than that in the malignant tumors (Plt;0.05). In all cases, the expression of ΔNP63 was higher than that of TAP63 (Plt;0.05); the difference in the expression of TAP63 among the groups was not significant (Pgt;0.05). Conclusion ΔNP63 in epithelial ovarian cancer is highly expressed, which may become the molecular makers with diagnosis and prognosis of epithelial ovarian cancer diagnosis in the future.