Analysis of the expression and association of retinoblastoma binding protein 6 with the JNK signaling pathway in prostate cancers.

This study aims to investigate the expression of retinoblastoma binding protein 6 (RBBP6) in prostate cancer (PCa) and its association with the c-Jun N-terminal kinase (JNK) pathway. Immunohistochemistry was used to detect RBBP6 and JNK1/2 expression in PCa and benign prostatic hyperplasia tissues. RBBP6 expression in PCa cells (LNCap, PC3, and DU145) and noncancerous prostate epithelial cells (RWPE-1) was determined by quantitative real-time polymerase chain reaction and western blot analysis. PC3 and DU145 cells were transfected with RBBP6 small interfering RNAs (siRNAs) to examine the biological characteristics. Anisomycin (a JNK activator) with/without RBBP6 siRNA was used to treat PC3 cells for further investigating the ramification of the RBBP6-mediated JNK pathway in PCa. PCa tissues and cells showed higher RBBP6 and JNK1/2 expression. RBBP6 was positively correlated with JNK1/2 in PCa tissues. Besides, RBBP6 expression was correlated to clinical tumor stage, lymph node metastasis, Gleason grade, preoperative prostate-specific antigen level, as well as prognosis of PCa. RBBP6 siRNA reduced cell proliferation, arrested cells at G2/M, and promoted cell apoptosis, and suppressed JNK pathway. In addition, migration and invasion decreased after the RBBP6 siRNA transfection with downregulated matrix metallopeptidase-2 (MMP-2) and MMP-9. Anisomycin promoted the proliferation, invasion, and migration of PC3 cells and inhibited PC3 cell apoptosis, which could be reversed by RBBP6 siRNA. RBBP6 expression was upregulated in PCa tissues and positively correlated with expression level of JNK1/2. With inhibition of RBBP6 expression, the proliferation, invasion, and migration of PCa cells decreased dramatically, while PC3 cell apoptosis increased appreciably, accompanied by the suppression of the JNK pathway.

Role of miR-23a/Zeb1 negative feedback loop in regulating epithelial-mesenchymal transition and tumorigenicity of intraocular tumors.

Role of the two-way negative feedback regulation channel formed by miR-23a and Zeb1 in epithelial-mesenchymal transition (EMT), tumorigenic ability, and migration and metastasis capacity of the intraocular malignant tumor cells was investigated. Molecular biological methods such as real time-quantitative PCR (RT-qPCR), immunoblotting method, and immunofluorescence were used to detect the expression levels of mRNA and protein in the Zeb1 factor in OCM-1, WERI-RB1, and Y79 cells before and after miR-23a transfection. Transwell cells were used to detect the in vitro membrane permeation and migration ability in OCM-1, WERI-RB1, and Y79 cells (non-transfection group, blank control transfection group, mimic transfection group, inhibitor transfection group). The results revealed that the relative expression of miR-23a in the cells in the miR-23a mimic transfection group increased significantly compared with that in the control group (p<0.05). There were significant differences in the relative expression of mRNA between the mimic transfection and control group (p<0.05). RT-qPCR detection showed that the relative expression of mRNA of the epithelial-labeled factor E-cadherin increased significantly in the miR-23a mimics group (p<0.05). Expression of the protein E-cadherin increased while the expression of the mesenchyme-labeled proteins of vimentin and N-cadherin decreased in the mimics group. Zeb1 has a negative feedback effect on miR-23a. They can form a negative feedback loop. The results showed that miR-23a and Zeb1 form a bidirectional inhibitory negative feedback loop, which plays an important role in regulating EMT. In conclusion, the significant changes in the mesenchymal phenotype of the stable strains with Zeb1 overexpressed in the OCM-1 cells cannot be completely explained with the changes in cytoskeleton caused by EMT.

Comparison of treatments for bullous keratopathy in rabbits.

The aim of the present study was to compare deep lamellar endothelial keratoplasty (DLEK) and penetrating keratoplasty (PK) treatments for bullous keratopathy (BK). In total, 36 healthy New Zealand white rabbits were randomly divided into 3 groups termed the experimental, DLEK and PK groups. The experimental control group received no treatment. The DLEK and PK groups were observed for corneal astigmatism at 1, 2, or 3 months post-surgery using a corneal topography instrument and a slit lamp microscope. The incidence of immune rejection after 3 months of recovery was determined using hematoxylin and eosin (H&E) staining. The corneal specimens from the surgery groups were compared with those from the control group. In the 12 rabbit eyes that underwent the DLEK surgery, the central cornea became clear after 1 week. After 3 months, these corneas were almost transparent and no eye infections or other complications were observed in 10 of the eyes, while surgical perforations in 2 eyes led to surgical lamellar failure. In the PK surgery group, in which 12 rabbit eyes were also treated, nine were almost transparent after 3 months of recovery, while three eyes were immunologically rejected due to the corneal grafts. The occur-rences of corneal astigmatism that were observed following DLEK and PK treatment were significantly different after 1, 2 and 3 months of recovery (P<0.05). Normal corneal staining was observed in the DLEK and PK rabbits subjected to H&E staining after 3 months of recovery. A BK animal model was established by curetting the Descemet's membrane (DM film). In comparison with PK, DLEK is a superior surgical treatment for BK.