Stabilization of 4E-BP1 by PI3K kinase and its involvement in CHK2 phosphorylation in the cellular response to radiation.

Objectives: 4E-BP1 is a family member of eIF4E binding proteins (4E-BPs) which act as the suppressors of cap-dependent translation of RNA via competitively associating with cap-bound eIF4E. RNA translation regulation is an important manner to control the cellular responses to a series of stress conditions such as ionizing radiation (IR)-induced DNA damage response and cell cycle controlling. This study aimed to determine the mechanism of 4E-BP1 stabilization and its potential downstream target(s) in the response to IR. Methods: PI3Ks kinase inhibitors were used to determine the signaling control of 4E-BP1 phosphorylation and protein stability. shRNA strategy was employed to silence the expression of 4E-BP1 in HeLa and HepG2 cells, and determine its effect on the irradiation-induced CHK2 phosphorylation. The protein degradation/stability was investigated by western blotting on the condition of blocking novel protein synthesis by cycloheximide (CHX). Results: The phosphorylation of 4E-BP1 at Thr37/46 was significantly increased in both HepG2 and HeLa cells by ionizing radiation. Depression of 4E-BP1 by shRNA strategy resulted in an incomplete G2 arrest at the early stage of 2 hours post-irradiation, as well as a higher accumulation of mitotic cells at 10 and 12 hours post-irradiation as compared to the control cells. Consistently, the CHK2 phosphorylation at Thr68 induced by IR was also attenuated by silencing 4E-BP1 expression. Both PI3K and DNA-PKcs kinase inhibitors significantly decreased the protein level of 4E-BP1, which was associated with the accelerated degradation mediated by ubiquitination-proteasome pathway. Conclusion: PI3K kinase activity is necessary for maintaining 4E-BP1 stability. Our results also suggest 4E-BP1 a novel biological role of regulating cell cycle G2 checkpoint in responding to IR stress in association with controlling CHK2 phosphorylation.

Comparison of anterior section parameters using anterior segment optical coherence tomography and ultrasound biomicroscopy in myopic patients after ICL implantation.

AIM: To compare the agreement of anterior chamber depth (ACD) and central vault measurements obtained by anterior segment optical coherence tomography (AS-OCT) and ultrasound biomicroscopy (UBM) of post surgical high myopic eyes with posterior chamber phakic intraocular lens (Visian ICL; STAAR Surgical) implantation. METHODS: Fifty-two phakic eyes of 28 high myopic patients who underwent implantable Collamer lens (ICL) surgery for the correction of high myopia were studied. The postoperative ACD, the distance between the corneal endothelium and the anterior surface of ICL (cornea-ICL) and the central vault were measured with the AS-OCT system and the UBM system. Intraclass correlation coefficient (ICC) and the Bland-Altman plot were used to evaluate the repeatability and agreement of two devices. RESULTS: The mean ACD, cornea-ICL and central vault in the 52 phakic eyes after ICL surgery was 3.19±0.28 mm, 2.47±0.28 mm, 0.50±0.19 mm by AS-OCT and 3.13±0.25 mm, 2.49±0.25 mm, 0.44±0.19 mm by UBM, respectively. Pairwise comparison of ACD and central vault measurements showed significant differences between AS-OCT and UBM (P<0.05). However, no statistically significant difference was found between these imaging techniques in cornea-ICL (P>0.05). The Pearson correlation coefficient (r) between AS-OCT and UBM measurements for ACD, cornea-ICL and vault was 0.88, 0.80 and 0.89, respectively (P<0.001). The ICC was 0.89-0.94 for the measurements of AS-OCT and UBM. Bland-Altman analysis showed the 95% limits of agreement of ACD, cornea-ICL, central vault measurements between these two devices were -0.20 to 0.32 mm, -0.36 to 0.32 mm and -0.12 to 0.24 mm, respectively. CONCLUSION: Central ACD and vault measurements using AS-OCT demonstrated a slight significantly higher value than using UBM in phakic eyes after ICL surgery. These two devices should not be used interchangeably for measurements of central ACD and vault in patients after phakic intraocular lens implantation.

Corneal topograph-guided laser subepithelial keratomileusis (LASEK) corrects decentered ablation after laser in situ.

PURPOSE: Corneal topograph-guided laser subepithelial keratomileusis (LASEK) can effectively correct decentered ablation occurring post laser in situ keratomileusis (LASIK) and to enhance our understanding and diagnosis of decentered ablation following LASIK. METHODS: Previous studies in the relevant literature are reviewed, and a case report is provided. RESULTS: A patient with high myopia undergoing LASIK in both eyes presented with distorted vision in the left eye, which interfered with the vision in the right eye and caused blurred vision in both eyes. The patient was unable to see objects with both eyes. After receiving corneal topography-guided LASEK, the signs of distorted vision in the left eye and bilateral blurred vision were significantly alleviated, and the patient could see objects with both eyes simultaneously. CONCLUSION: Clinical ophthalmologists should be aware of the occurrence of decentered ablation after LASIK. Corneal topography-guided LASEK is an efficacious tool for correcting decentered ablation.