1.Expressions of CD133 in hepatocellular carcinoma and its clinical significance
Hua ZHANG ; Zhihong GAO ; Tianying XU ; Jianfeng ZHU ; Yaug ZHANG
Cancer Research and Clinic 2009;21(7):440-443
Objective To investigate relationship between CD133 expression and progression of hepatocellular carcinoma (HCC) by detecting expressions of CD133. Methods 65 cases with HCC were enrolled and their HCC tissues and peripheral blood samples were taken. Expression of CD133 in HCC tissue was detected with immunohistochemical method and the percentages of CD45- CD133+ cells in peripheral blood were assayed by the method of flow cytometry (FCM), and 10 healthy persons were chosen as controls. Results In HCC patients, the positive expression rate of CD133 in HCC tissues was 6.4 % and the percentage of CD45- CD133+ cells in peripheral blood was 2.0 %. The expression rate of CD133 was not found in normal liver tissues and peripheral blood (P <0.01). The amount of CD133+ cells was significantly related to portal vein tumor embolus (P<0.01), tumor amicula integrality (P<0.01), pathology grade (P<0.01) and TNM stage (P <0.01), but not related to tumor diameter(P0.05) and AFP value (P0.05). It showed a positive correlation between HCC tissues and peripheral blood (r=0.36, P <0.01). Conclusion The percentage of CD133+ cell in HCC tissues and peripheral blood from HCC patient is greatly correlated to the progression of HCC. It is useful to exam the CD133 expression in predicting the diagnosis and prognosis evaluation.
2.Suppressing effects of RNA interference targeting AKT1 and phosphatidylinositol 3-kinase p85 on the invasion ability of malignant glioma U251 cells in vitro
Lei HAN ; An-Ling ZHANG ; Xiao YUE ; Yaug YANG ; Guang-Xiu WANG ; Zhi-Fau JIA ; Pei-Yu PU ; Chun-Sheng KANG
Chinese Journal of Neuromedicine 2010;9(6):566-570
Objective To investigate the suppressing effects of RNA interference (RNAi)targeting AKT1 and phosphatidylinositol 3-kinase p85 (PI3KP85) on the invasion ability of malignant glioma U251 cells in vitro.Methods Normal control group,negative control group (U251 cells being transfected by nonsense sequence of adenovirus) and gene treatment group were chosen in the experiment.A recombinant rctrovims expressing short interference RNA (siRNA) sequence targeting AKTi and PI3KP85 genes was established and transfected into the U251 cells in the gene treatment group.The silencing effect of RNAi on AKTI and PI3KP85 expressions was identified by real time PCR and Westem blotting,respectively.Western blotting was also employed to analyze the expression of some functional proteins.Enzyme-linked immuno sorbent assay (ELISA) was used to detect the changes of concentration of ectocytic matrix metalloproteinases 2 (MMP2) and MMPg.The invasion ability of U251 cells in the 3 groups was evaluated by scarification and Transwell assay.Results The expressions of AKT1 and PI3KP85 in U251 cells in the gene treatment group were dramatically down-regulated.As compared with the normal control and negative control groups,the gene treatment group showed significantly lower expression level of MMP2 and MMP9(P<0.05);meanwhile tissue inhibitor of matrix metalloproteinase-2(TIMP2)in the gene treatment group was significantly up-regulated.ELISA also indicated obvious changes of concentration of ectocytic MMP2 and MMP9.The scarification and Transwell assay showed that the invasion ability in the gene treatment group was significantly decreased as compared with that in the other 2 groups (P<0.05).Conclusion RNAi targeting AKT1 and PI3KP85 can significantly down-regulate the expressions of AKT1 and PI3KP85 and decrease the invasion ability of U251 cells in vitro.