ObjectiveTo investigate the expression of microRNA (miRNA, miR)-141-3p in hepatocellular carcinoma (HCC) tissues and its effect on proliferation and invasion of HCC.MethodsBioinformatics tools were used to predict putative miRs with search potential downstream gene–GP73, hsa-miR-141-3p was selected for further analysis and observations. A dual-luciferase reporter assay was performed to validate GP73 as a target of hsa-miR-141-3p. Real time PCR (qRT-PCR) or Western blot was performed to measure the expression level of miR-141-3p and GP73 in HCC and adjacent tissue. MTT, EdU, and Transwell were used to measure cell proliferation and migration.ResultsIt was identified that the expression level of miR-141-3p was significantly lower in the HCC tissues than that of adjacent tissues (P<0.05), but different in GP73 mRNA and its protein (P<0.05). Clinical analysis indicated that decreased miR-141-3p expression was significantly correlated with advanced tumor grade, advanced TNM stage, and vascular invasion (P<0.05). MTT assay showed that the absorbance (A) value of Huh-7 cells transfected with miR-141-3p overexpression plasmid was significantly decreased compared with the miR-negative control (NC) group (transfected empty plasmid) and the blank control group (P<0.05).The EdU detection results showed that the ratio of EdU positive cells in Huh-7 and MHCC-97H cells were lower than those in the blank control group and the miR-NC group after transfection of miR-141-3p (P<0.05). Transwell test found that after transfection of miR-141-3p, the number of invading cells and the number of migrated cells in MHCC-97H cells were lower than those in the blank control group and the miR-NC group (P<0.05). The results of cytological function recovery experiments showed that the number of invading cells and the number of migrated cells in MHCC-97H cells of miR-141-3p+GP73 group were lower than those in blank control group and miR-NC group (P<0.05), but higher than those of the miR-141-3p group and miR-141-3p+GP73-NC group (P<0.05).ConclusionsThe present study revealed that miR-141-3p is reduced in HCC tissues. Additionally, GP73 expression levels are negatively correlated to miR-141-3p in HCC tissues, and that is associated with the progression of HCC. Overexpression of miR-141-3p significantly inhibit proliferation, invasion, and migration of HCC cells.Reversal of partial inhibition of miR-141-3p can be achieved by restoring the expression of the GP73 gene without the 3′ untranslated region (UTR).
Objective To observe the effect of intravitreal injection of bevacizumab (Avastin, IVB) on the expression of integrin-linked kinase (ILK) in fibrovascular membranes and the number of vascular endothelial cells (VECs) in proliferative diabetic retinopathy (PDR). Methods Twenty-four fibrovascular membrane samples were collected during pars plana vitrectomy in 24 patients with PDR. 12 PDR patients had received a single 1.25 mg IVB 7 days preoperatively (bevacizumab group), the other 12 patients (non-bevacizumab group) had not received IVB. For each of 24 fibrovascular membranes specimen, the number of VECs in the membranes were counted after staining with hematoxylin-eosin and von willebrand factor. Expressions of ILK in the fibrovascular membranes were detected through immunohistochemistry analysis. Results Immunohistochemistry revealed that ILK was highly expressed in all of 24 fibrovascular membranes of PDR.The average optical density of ILK expression level in bevacizumab and non-bevacizumab group were (127.78plusmn;15.08) and (129.03plusmn;16.26) respectively, the difference was not statistically significant (t=0.330，P=0.745).The number of VECs in fibrovascular membranes in bevacizumab and non-bevacizumab group were 21.50plusmn;3.94 and 41.33plusmn;7.44 respectively, the difference was statistically significant (t=3.872，P=0.003). Conclusions ILK was expressed in fibrovascular membranes of PDR. IVB can decrease the number of VECs during the process of PDR, but it can not affect the expression of ILK protein.