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Objective:To explore the effects of long noncoding RNA adenosine diphosphate-dependent glucokinase antisense RNA 1(ADPGK-AS1) on the proliferation, migration and invasion of human retinoblastoma (RB) Y-79 cells and its regulatory effect on microRNA-623 (miR-623).Methods:The peritumoral tissue and RB specimens were collected from 39 eyes of 39 patients with RB during surgery in The First Affiliated Hospital of Zhengzhou University and Zhumadian Central Hospital from February 2017 to November 2018.Real-time fluorescence quantitative PCR was employed to detect the expression of ADPGK-AS1 and miR-623 in the specimens.Human RB line Y-79 cells were cultured in vitro and divided into small interfering RNA-normal control (siRNA-NC) group, siRNA-ADPGK-AS1 group, microRNA (miR)-NC group, miR-623 group, siRNA-ADPGK-AS1+ anti-miR-NC group and siRNA-ADPGK-AS1+ anti-miR-623 group.The cell proliferation rate was detected by MTT method.Transwell cell experiment was performed to detect the number of migrating and invading cells.The dual luciferase reporter experiment was used to evaluate the targeting relationship between ADPGK-AS1 and miR-623.The expression of Ki-67, matrix metalloproteinases (MMP)-2, and MMP-9 in the cells was detected by Western blot assay.Written informed consent was obtained from each patient prior to any medical examination and treatment.This study protocol adhered to the Declaration of Helsinki.The use of the human specimens was approved by an Ethics Committee of The First Affiliated Hospital of Zhengzhou University (No.2017-KY-73). Results:Compared with the peritumoral tissue, the relative expression level of ADPGK-AS1 in the RB tissue was significantly increased, and the relative expression level of miR-623 was significantly reduced ( t=40.522, 48.497; both at P<0.01). Compared with the siRNA-NC group, both the relative expression level of Ki-67 protein and the proliferation A value of RB Y-79 cells were significantly reduced in the siRNA-ADPGK-AS1 group ( t=26.833, 18.522; both at P<0.01). The relative expression levels of MMP-2 and MMP-9 proteins in the siRNA-ADPGK-AS1 group were significantly lower than those in the siRNA-NC group ( t=22.123, 26.183; both at P<0.01). The number of migrating and invading cells in the siRNA-ADPGK-AS1 group was significantly less than that in the siRNA-NC group ( t=12.385, 19.201; both at P<0.01). The dual luciferase report experiment confirmed that ADPGK-AS1 targeted miR-623.The protein expression levels of the Ki-67, MMP-2 and MMP-9 in the miR-623 group were significantly lower than those in the miR-NC group ( t=22.137, 22.200, 21.094; all at P<0.01). Compared with the miR-NC group, the proliferation A value of Y-79 cells in the miR-623 group was significantly lower, and the number of migrating and invadoing cells was significantly less ( t=16.398, 11.400, 17.846; all at P<0.01). The relative expressions levels of Ki-67, MMP-2 and MMP-9 proteins in the siRNA-ADPGK-AS1+ anti-miR-623 group were significantly higher than those in the siRNA-ADPGK-AS1+ anti-miR-NC group ( t=20.795, 17.493, 23.479; all at P<0.01). Compared with the siRNA-ADPGK-AS1+ anti-miR-NC group, the proliferation A value of Y-79 cells in the siRNA-ADPGK-AS1+ anti-miR-623 group was significantly increased ( t=15.600, P<0.01), and the number of migrating and invading cells was obviously elevated ( t=14.495, 17.855; both at P<0.01). Conclusions:Knockdown of ADPGK-AS1 gene can inhibit the proliferation, migration and invasion of Y-79 cells by up-regulating the expression of miR-623.
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Objective:To investigate whether microRNA-338-3p (miR-338-3p) affects the proliferation and apoptosis of choroidal microvascular endothelial cells by regulating the expression of T cell factor 4 (TCF4).Methods:Human choroidal microvascular endothelial cells were cultured in vitro, and they were divided into vascular endothelial growth factor (VEGF) group and Normal control group.The Cultured cells in the VEGF group were divided into four subgroups, and were transfected with miR-NC, miR-338-3p mimics, miR-338-3p mimics + pcDNA, miR-338-3p mimics + pcDNA-TCF4 before VEGF treatment, respectively.Quantitative real-time PCR (qRT-PCR) was used to detect the expression of miR-338-3p and TCF4.MTT assay was used to detect cell proliferation.Flow cytometry was used to detect the apoptosis rate.The double luciferase report experiment verified the targeting relationship of miR-338-3p and TCF4.Western blot was used to detect the expression of Ki-67, PCNA, bax, and bcl-2. Results:After VEGF treatment, the expression level of miR-338-3p was significantly reduced, the expression level of TCF4 mRNA was significantly increased, the cell viability was significantly increased, and the apoptosis rate was significantly decreased, the levels of Ki-67, PCNA, bcl-2 protein were increased significantly, and the level of bax protein was decreased significantly (all at P<0.05). After miR-338-3p overexpression, the cell viability was significantly reduced, the apoptosis rate was significantly increased, and the levels of Ki-67, PCNA, and bcl-2 proteins were significantly reduced, the level of bax protein was increased significantly (all at P<0.05). Double luciferase reporting experiments confirmed that miR-338-3p targeted TCF4.Compared with the VEGF + miR-338-3p + pcDNA group, the cell viability of the VEGF + miR-338-3p + pcDNA-TCF4 group was significantly increased ([56.48±13.20]% vs. [96.24±16.24]%), and the apoptosis rate was significantly reduced ([30.59±3.57]% vs.[12.36±1.29]%), the levels of Ki-67, PCNA and bcl-2 protein were significantly increased (0.41±0.11 vs. 0.96±0.19; 0.44±0.10 vs. 0.97±0.20; 0.55±0.12 vs. 0.98±0.15), and the levels of bax protein were significantly decreased (0.87±0.13 vs. 0.42±0.11) ( t=5.700, 14.408, 7.516, 7.111, 6.715, 7.927; all at P<0.01). Conclusions:Overexpression of miR-338-3p can negatively regulate TCF4 expression, thereby inhibite choroidal microvascular endothelial cell proliferation and induce apoptosis.
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Background Orbital cavernous hemangiomas (OCH) is a common benign orbital tumor in adult,and accurate localization and diagnosis before operation supports the significant premise for surgical safety and success of tumor extraction.Objective This study was to research the clinical characteristics,preoperative diagnosis,the selection for different surgical approaches,therapeutic effectiveness and complication prevention of OCH.Methods The clinical data of 117 eyes of 117 patients with OCH who received surgery were retrospectively analyzed.The patients received surgery in Henan Eye Institute,Henan Eye Hospital from January 2011 to December 2014 and followed-up for 3 months to 5 years.The visual acuity,exophthalmos,ocular movement,orbital A/B ultrasound,color Doppler image,CT and MRI were examined before and after surgery.Results The primary clinical manifestations of OCH were gradual exophthalmos and impaired vision.The accordance rate of preoperative diagnosis and pathological diagnosis was 100％ in the group of patients.The surgical approachs included conjunctival approach in 52.14％ (61/117),lateral orbitectomy in 30.77％ (36/117),anterior approach in 16.24％ (19/117) and lateral combined with medial approach in 0.85％ (1/117).At the end of followed-up,visual acuity was significantly improved in 30.77％ (36/117),declined in 8.55％ (10/117) and unchanged in 60.68％ (71/117).Temporary complications after surgery were pupil dilatation in 14.53％ (17/117),emorrhoea in 1.71％ (2/117),ocular motility disorders in 16.24％ (19/117) and ptosis in 4.27％ (5/117).The permanent complications after operation were pupil dilatation in 2.56％ (3/117),visual loss in 0.85％ (1/117) and permanent abduction imitation in 0.85％ (1/117).Conclusions Accurate qualitative and site-specific diagnosis and correct choice of surgeries for OCH depend on clinical and iconographical examinations.Suitable surgical approach and operative skill are helpful to the therapeutic outcome and safety of OCH.
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Background Orbital blow-out fracture often results in the abnormalities of appearance and function of eye.Because of the special and complex anatomical structure of orbital cavity,it is difficult to design and manufacture the corresponding orbital implants.The computer-aided designing and manufacturing (CADM) technology provides a new approach to orbital implants.However,the clinical value of this method is still under evaluation.Objective This study was to investigate the application and the therapeutic effect of CADM for orbital blow-out fracture.Methods The clinical data of 74 eyes of 74 patients who received surgery for orbital blow-out fracture from July 2006 to July 2012 in Henan Eye Institute,Henan Eye Hospital were retrospectively analyzed.Fiftyeight patients underwent CADM implanted surgery and 16 patients received non-CADM surgery in the same period with matched age,gender and lateral eyes in both groups.The individualized 3D orbital implants were designed and manufactured by the technology of CADM and then were implanted in the bone defects in the CADM group,while the traditional hydroxyapatite artificial bone or high density porous polyethylene material (Medpor) was utilized in the non-CADM group with a fellow-up duration for 22 to 69 months.The best corrected visual acuity (BCVA),eyeball exophthalmos,ocular position,eye movement,diplopia and postoperative complications were evaluated.Results The preoperative BCVA were 0.71±0.37 and 0.69±0.41,and the postoperative BCVA were 0.74±0.38 and 0.72±0.41 in the CADM group and the non-CADM group,respectively,showing an insignificant intergroup difference (Fgroup =0.043,P=0.837),but a significant variation was found over time (Ftime =13.576,P ＜ 0.01).The BCVA was significantly improved after surgery compared with before surgery in both groups (both at P＜0.05).No significantdifferences were found in the number of eyes with curative and improved diplopia and eye movement disorders between the two groups during the fellow-up duration (Z =-0.298,P =0.766;Z =-0.548,P =0.584).The preoperative eyeball exophthalmos values were (3.93±0.99)mm and (3.88±0.97)mm and the postoperative ones were (0.91 ±0.67)mmand (1.84±0.80) mm in the CADM group and the non-CADM group,respectively,without significant differencebetween the two groups (Fgroup =3.558,P =0.063).However,the eyeball exophthalmos values after operation wereremarkably lower than those before operation in both groups (both at P＜0.05).CT imaging displayed implants fitting well with fracture defect and attached to bone tissue accurately in all of the eyes in the CADM group,but in the nonCADM group,the bulge of implants damaging extraocular muscles or optical nerve was found in 2 eyes.No postoperative complication was seen throughout the fellow-up duration in the CADM group.Conclusions CADM technology for orbital blow-out fracture can reconstruct a 3D bony orbit and effectively repair ocular position and appearance,and furthermore restore eye movement and visual functions.The therapeutic outcome of CADM technology for orbital blow-out fracture is superior to conventional implants.