PubMed | Sun Yat Sen University, Kiang Wu Hospital Macau and The First Peoples Hospital of Shunde Foshan
Type: Journal Article | Journal: International journal of clinical and experimental medicine | Year: 2015
This study was aimed to investigate the role of miR-29a in myocardial cell apoptosis induced by high glucose. Myocardial cells were cultured in normal (5.6 mmol/l) or high glucose medium (30 mmol/l). The apoptotic rate of myocardial cells was evaluated using flow cytometry. The mRNA levels of Bax, Bcl-2, miR-29a, and IGF-1 were determined using real-time quantitative PCR (RT-qPCR). The level of IGF-1 in the culture medium was analyzed using enzyme-linked immunosorbent assay (ELISA). The interaction sites between miR-29a and IGF-1 was analyzed using the the Targetscan program. The regulatory effect of miR-29a on the expression of IGF-1 was investigated using dual luciferase reporter system. The results showed that the expression of miR-29a and the Bax/Bcl-2 ratio in myocardial cells were significantly increased after the cells were cultured in high glucose medium for 72 h, which was consistent with increased apoptosis of myocardial cells. The expression of IGF-1 in myocardial cells was significantly decreased after the cells were cultured in high glucose medium for 72 and 96 h. Targetscan identified a potential binding site on the 3-UTR of IGF-1 for miR-29a. We also observed that miR-29a mimic and miR-29a inhibitor reduced and increased the expression of IGF-1 in myocardial cells cultured in high glucose medium, respectively. Dual luciferase reporter analysis showed that miR-29a significantly reduced the fluorescence intensity of wild-type psichek2-IGF-1-3UTR-WT but the fluorescence intensity of mutant psichek2-IGF-1-3UTR-MT was not significantly affected. In conclusions, the expression of miR-29a in myocardial cells cultured in high glucose medium was significantly increased, which down-regulated IGF-1 and increased myocardial cell apoptosis.
Wang F.-H.,Guangzhou Women and Childrens Medical Center |
Wen J.-M.,Kiang Wu Hospital Macau |
Vong H.-T.,Guangzhou Women and Childrens Medical Center |
Yip Y.-C.,Guangzhou Women and Childrens Medical Center
Chinese Journal of Pathology | Year: 2013
Objective To explore the expression and diagnostic significance of glypican-3 ( GPC3 ) in hepatoblastoma. Methods Five tissue microarray paraffin blocks were constructed to include 54 cases of hepatoblastoma. The tumor tissue samples were obtained from 3 surgical biopsies, 33 needle biopsies, 5 stage 1 resection tumors, and 13 stage II resection tumors after transcatheter arterial chemoembolization. Ten samples of non-neoplastic hepatic tissue adjacent to tumor were used as control. Immunohistochemical staining of GPC3 (clone 1G12) was performed. Among the 54 cases of hepatoblastoma, 22 cases were fetal subtype, 24 cases were mixed fetal and embryonal subtype and 8 cases were mixed epithelial and mesenchymal type. Results GPC3 was positive in fetal epithelial cells (54/54,100%), but negative or weakly positive in embryonic epithelial cells in all cases of hepatoblastoma. Undifferentiated small cells and all mesenchymal components were negative for the expression. Non-neoplastic hepatocytes adjacent to tumor were negative for GPC3 expression (0/10 ). Conclusions Fetal epithelial components of hepatoblastoma express GPC3 protein detectable by immunohistochemistry. Normal hepatocytes after birth, small cell undifferentiated and embryonic epithelial components of hepatoblastoma do not or weakly express GPC3 protein. Therefore, GPC3 immunohistochemistry offers a valuable aid to the diagnosis of hepatoblastoma in infants and children.