title: Down-Regulation of MicroRNA-133b Suppresses Apoptosis of Lens Epithelial Cell by Up-Regulating BCL2L2 in Age-Related Cataracts.
abstract: BACKGROUND MicroRNA-133b (miR-133b) has been reported to be involved in many diseases, including ovarian cancer and osteosarcoma. Accumulating evidence suggests that miR-133b plays important roles in human disease. In this study, we aimed to investigate the molecular mechanism, including the potential regulator and signaling pathways, of BCL2L2. MATERIAL AND METHODS We first searched the online miRNA database (www.mirdb.org) using the "seed sequence" located within the 3'-UTR of the target gene, and then performed luciferase assay to test the regulatory relationship between miR-133b and BCL2L2. Western blot and real-time PCR were used to determine the expression of BCL2L2 in human samples or cells treated with miRNA mimics or inhibitors. Flow cytometry was conducted to evaluate the apoptosis status of the cells. RESULTS We validated BCL2L2 to be the direct gene using a luciferase reporter assay. We also conducted real-time PCR and Western blot analyses to study the mRNA and protein expression level of BCL2L2 among different groups (control: n=29, cataract: n=33) or cells treated with scramble control, miR-133b mimics, BCL2L2 siRNA, and miR-133b inhibitors, and identified the negative regulatory relationship between miR-133b and BCL2L2. We also conducted experiments to investigate the influence of miR-133b and BCL2L2 on the viability and apoptosis of cells. The results showed that miR-133b positively interfered with the viability of cells, while BCL2L2 negatively interfered with the viability of cells, and that miR-133b inhibited apoptosis while BCL2L2 accelerated apoptosis. CONCLUSIONS BCL2L2 was the virtual target of miR-133b, and we found a negative regulatory relationship between miR-133b and BCL2L2. MiR-133b and BCL2L2 interfered with the viability and apoptosis of cells.
authors: Department of Ophthalmology, The People's Hospital of Beizhou, Binzhou, Shandong, China (mainland).
authors: Feng
authors: F
authors: Zhang
authors: Department of Ophthalmology, The People's Hospital of Binzhou, Binzhou, Shandong, China (mainland).
authors: Weizhe
authors: W
authors: Meng
authors: Department of Ophthalmology, The People's Hospital of Binzhou, Binzhou, Shandong, China (mainland).
authors: Bin
authors: B
authors: Tong
mesh_terms: D020413:3' Untranslated Regions; D017209:Apoptosis / Q000235:genetics; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D002386:Cataract / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D015536:Down-Regulation; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D006801:Humans; D007908:Lens, Crystalline / Q000378:metabolism; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction; D015854:Up-Regulation
chemical_list: D020413:3' Untranslated Regions; D051017:Apoptosis Regulatory Proteins; C491073:BCL2L2 protein, human; C548234:MIRN133 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D034741:RNA, Small Interfering