Optic nerve injury induces rapid retinal neurodegeneration; however, how distinct retinal cell type responses are coordinated from the hyperacute injury phase to the early repair phase remains incompletely understood. In this study, to explore the dynamic changes in intercellular and intracellular signaling events between different cell types and elucidate their potential roles in retinal ganglion cell survival and early repair, we generated a time-resolved single-nucleus RNA sequencing atlas of adult male mouse retinas across five hyperacute-to-acute timepoints (2 hours, 8 hours, 1 day, 3 days, and 7 days) following optic nerve injury. Using computational network analysis, we reconstructed dynamic cell-to-cell communication and subsequent internal genetic responses among retinal ganglion cells, Müller glia, microglia, and endothelial cells. Distinct stage-specific intercellular communication networks were identified, including transient Itgb1-associated signaling between Müller glia and retinal ganglion cells that peaked at early timepoints, enhanced Nrxn1-Nlgn1-mediated signaling in endothelial cells during the acute phase, and sustained Sema6a-Plxna4 interactions in microglia through day 7. Functional pathway analysis linked these signaling events to focal adhesion, energy metabolism, immune regulation, and cell adhesion pathways. Multiplex immunofluorescence further validated the temporal dynamics and spatial localization of key signaling molecules, including Itgb1, Nlgn1, and Plxna4, consistent with the transcriptomic findings. Collectively, these results delineate a coordinated hyperacute-to-acute neuro-glial-vascular signaling network that supports retinal ganglion cell survival and identify potential molecular targets for therapeutic intervention following optic nerve injury.

Introduction Retinoblastoma is divided into two clinical types, sporadic and heritable, and is the most common primary intraocular malignancy in infancy and childhood. It was the first malignancy to demonstrate the genetic etiology of cancer, with the RB1 gene as the only pathological gene present in heritable cases. Although the RB1 mutation p.E125* had been previously reported in other retinoblastoma patients, it lacked functional analysis.Methods In this study, we identified the RB1 p.E125* mutation in a bilateral retinoblastoma patient from China. We investigated the distribution, cell localization, and function of this mutation using molecular biology and structural analysis following the transfection of cells with plasmids encoding the mutant RB1 gene.Results Functional analyses revealed abnormal protein localization, altered cell cycle distribution, and apoptosis in cells transfected with the mutant RB1 plasmids.Discussion Our findings contribute to a better understanding of RB1 mutation hotspots. Furthermore, our results highlight the importance of offering targeted genetic testing and counseling to families with RB1 mutations. The identification of the somatic origin of this mutation was vital in ruling out the heritability of this condition in this specific patient.

PurposeTo evaluate the efficiency and safety profiles of a technique of simultaneous reconstruction of traumatic lacrimal duct obstruction and medial canthus deformity using micro-anchors.MethodsA retrospective interventional study was performed from 2018 to 2022 on 26 consecutive patients who presented with medial canthal deformities and traumatic nasolacrimal duct obstruction. All patients underwent medial canthoplasty using micro-anchors combined with dacryocystorhinostomy to address the traumatic nasolacrimal duct obstruction. Preoperative and postoperative measurements of telecanthus, the superior or the inferior displacement of the medial canthal tendon (MCT), surgical technique of MCT fixation, advantages and limitations of using micro-anchors, functional and cosmetic outcomes were recorded and compared.ResultsSatisfactory and symmetrical medial canthal fixation outcomes were achieved in 96% (25/26) patients using the micro-anchors. The surgical technique was easy and offered secure and reliable reattachment of the MCT in long-term. The DCR surgery was anatomically and functionally patent in 92% (24/26) patients. One patient had unsatisfactory medial canthal outcomes where the canthus remained partly lateral, and inferior compared to the uninjured side. The two failures following DCR surgery were secondary to complete cicatricial closure and had persistent epiphora.ConclusionThe present study noted that micro-anchors can correct the medial canthal deformity and leave more space for the DCR osteotomy simultaneously.

Objective To correlate and evaluate the power and limitations of CT-DCG in determining the level and type of lacrimal duct obstruction in comparison to dacryoendoscopy in patients clinically suspected to be having partial or complete primary acquired nasolacrimal duct obstruction (PANDO). Methods A retrospective chart review was performed on 1232 lacrimal drainage systems of 957 patients who suffered from primary acquired nasolacrimal duct obstruction (PANDO) at Shanghai Ninth People's Hospital. Patients were examined with CT-DCG and correlated with dacryoendoscopy and the findings of clinical examination. Results Of the studied patients, 173 were men and 784 were women with an age range of 18–93 years. Of the 1232 lacrimal pathways, good CT-DCG images could be obtained in 980 cases and dacryoendoscopy in 957 cases. Of these complete obstructions were noted in 81% (794/980), and partial obstructions were identified in 19% (186/980) with CT-DCG. CT-DCG and dacryoendoscopy showed 68.4% agreement for the type of the obstruction and 63% for the level of the obstruction. The majority of the obstructions occurred at the sac-duct junction (62.5%) followed by the upper half of the nasolacrimal duct (27.5%). There was a significant difference in the correlation of the obstruction type with age group and with the duration of symptoms. As the duration of symptoms increased, the proportion of complete lacrimal duct obstructions as shown on CT-DCG images increased and the proportion of incomplete obstruction decreased ( p = 0.015). Conclusions The junction of lacrimal sac and nasolacrimal duct was the most common obstruction site. Age and the duration of symptoms influenced the type of obstruction noted. The degree and level of agreement between the investigations was moderate. A combination of CT-DCG and Dacryoendoscopy could together identify the location more accurately.

PURPOSE. The aim of this study was to describe the transcriptional changes of individual cellular components in the lacrimal sac in patients with primary acquired nasolacrimal duct obstruction (PANDO) and attempt to construct the first lacrimal sac cellular atlas to elucidate the potential mechanisms that may drive the disease pathogenesis. METHODS. Lacrimal sac samples were obtained intra-operatively during the endoscopic dacryocystorhinostomy (EnDCR) procedure from five patients. Single -cell RNA sequencing was performed to analyze each individual cell population including epithelial and immune cells during the early inflammatory and late inflammatory phases of the disease. RESULTS. Eleven cell types were identified among 25,791 cells. T cells and B cells were the cell populations with the greatest variation in cell numbers between the two phases and were involved in immune response and epithelium migration -related pathways. The present study showed that epithelial cells highly expressed the genes of senescenceassociated secretory phenotype (SASP) and were involved in influencing the inflammation, neutrophil chemotaxis, and migration during the late inflammatory stage. Enhanced activity of CXCLs-CXCRs between the epithelial cells and neutrophils was noted by the cell -cell communication analysis and is suspected to play a role in inflammation by recruiting more neutrophils. CONCLUSIONS. The study presents a comprehensive single -cell landscape of the lacrimal sac cells in different phases of PANDO. The contribution of T cells, B cells, and epithelial cells to the inflammatory response, and construction of the intercellular signaling networks between the cells within the lacrimal sac has further enhanced the present understanding of the PANDO pathogenesis.

PurposeTo evaluate the outcomes of endoscopy-assisted modified Weber-Ferguson's approach in the management of primary lacrimal sac tumors with extension into the neighboring tissues.MethodsA retrospective interventional study was performed on all patients with lacrimal sac tumors treated with the endoscopy-assisted modified Weber-Ferguson approach between January 2010 and June 2022 at the Shanghai Ninth People's Hospital, China. Data assessed include demographics, clinical presentations, imaging features, surgical techniques, histopathology, adjuvant modalities of management, complications, and outcomes.ResultsA total of 13 patients were included in the analysis. Epiphora and palpable mass lesion were the presenting complaint in 84.6% (11/13) of the patients. Nearly half of the patients (46.1%, 6/13) were misdiagnosed as lacrimal duct obstruction. All the lacrimal sac tumors in the present series showed uneven enhancement on T1-weighted MRI imaging. Postoperatively, 84.6% (11/13) patients recovered well with excellent esthetics and were disease-free after a mean follow-up of 58.6 months. Two patients who underwent additional exenteration developed recurrence and succumbed (at 41 and 96 months follow up) while they were on palliative chemoradiation.ConclusionThe endoscopic-assisted modified Weber-Fergusson surgical approach is effective in providing better visibility and accessibility to lacrimal sac tumors with extension into neighboring tissue.

Purpose: The purpose of this study was to evaluate the clinical features, surgical treatments, and outcomes of pediatric post-traumatic nasolacrimal duct obstructions and predictors for surgical outcomes. Methods: A retrospective chart review was performed of patients under the age of 18 years with the diagnosis of traumatic nasolacrimal duct obstructions during an 11-year period from a tertiary referral center. The clinical data were reviewed and analyzed. A binary logistic regression was performed to establish a predictive model for surgical failure after treatment. Results: Eighty-three eyes of 80 consecutive patients were included in the study. Anterior displacement of the lacrimal sac was noted in 71.1% (59/83) cases. Medial canthus deformities were observed in 79.5% (63/83) cases. External dacryocystorhinostomy and endoscopic dacryocystorhinostomy were performed on 65 and 18 cases, respectively. At a mean follow-up of 28 months following stent removal, anatomical and functional success rate was noted in 83.1% (69/83). The presence of orbital fractures involving the rim, gross skin scaring, lacrimal sac displacement, and fracture fragments in the vicinity of the lacrimal sac were found to be significant predictors for surgical failure. The area under the raw current curves of the predictive model was 0.888. Conclusion: This large series of post-traumatic pediatric nasolacrimal duct obstructions showed that sac-duct junction was the most common site of obstruction and anterior displacement of the lacrimal sac and bony fragments in the lacrimal sac area were significant predictors of failures. Medial canthal deformities can be addressed simultaneously with good outcomes.

OBJECTIVE:To evaluate the myocutaneous sliding flap for restructuring the eyelid of divided nevus, thus optimizing the aesthetic outcome.DESIGN:A retrospective clinical study.PARTICIPANTS:Clinical data from patients with a small or medium-sized divided nevus of the eyelids between January 2015 and December 2018 in the Shanghai Ninth People's Hospital were reviewed.METHODS:The safety and efficacy of a surgical approach using a myocutaneous sliding flap were evaluated based on postoperative features and complications.RESULTS:This study included 53 patients with an average age was 21.6 years (range, 2-68 years). The lesions involved the inner canthus in 5 patients (9.43%) and the lateral canthus in 14 patients (26.42%). Three (5.66%) patients had undergone primary surgery elsewhere. Overall, the eyelid margins were in good shape postoperatively at an average follow-up of 22.7 months, and there were no obvious eyelid deformities, lagophthalmos, or other unacceptable complications postoperatively. The nevi were benign intradermal (60.38%), junctional (16.98%), and compound types (22.64%) without malignant transformation, as confirmed by pathologic examination. No malignant transformation was observed until the end of the follow-up period.CONCLUSION:A myocutaneous sliding flap can provide an appropriately size pedicle graft and achieve satisfactory cosmetic results for divided eyelid nevus.

ObjectiveTo investigate the critical prognostic factors of patients with traumatic optic neuropathy (TON) treated with endoscopic transnasal optic canal decompression (ETOCD) and to perform multimodal analysis based on imaging examinations of optical coherence tomography angiography (OCTA) and CT scan. Subsequently, a new prediction model was established.MethodsThe clinical data of 76 patients with TON who underwent decompression surgery with the endoscope-navigation system in the Department of Ophthalmology, Shanghai Ninth People's Hospital from January 2018 to December 2021 were retrospectively analyzed. The clinical data included demographic characteristics, reasons for injury, interval between injury and surgery, multimode imaging information of CT scan and OCTA, including orbital fracture, optical canal fractures, vessel density of optic disc and macula, and the times of postoperative dressing change. Binary logistic regression was used to establish a model for best corrected visual acuity (BCVA) after treatment as a predictor of TON outcome.ResultsPostoperative BCVA improved in 60.5% (46/76) patients and did not improve in 39.5% (30/76) patients. The times of postoperative dressing change had a significant impact on the prognosis. Other factors affecting the prognosis were microvessel density of the central optic disc, the cause of injury, and the microvessel density above the macula. The area under the raw current curves of the predictive model was 0.7596.ConclusionsThe times of dressing changes after the operation, i.e., continuous treatment, is the key factor affecting prognosis. The microvessel density in the center of the optic disc and superior macula, quantitatively analyzed by OCTA, is the prognostic factor of TON and may be used as a prognostic marker of TON.

Purpose Divided nevus with verrucous hyperplasia will always recur after surgery but non-verrucous divided eyelid nevus rarely recur. This study analyzed the differential expression of Ki-67, S100, Melan A and HMB45 and identified the correlation between the clinical and histopathological features of verrucous and non-verrucous divided eyelid nevus. Methods This study included 29 patients, of whom 8 patients had verrucous divided nevus. Immunohistochemistry labeling was used to assess the expression of Ki-67, S100, Melan A and HMB45 after excision. The difference between verrucous and non-verrucous divided eyelid nevus was analyzed. Results The patients’ ages ranged from 2 to 59 years, with a mean age of 19 years. The lesion size ranged from 1.5 to 2.0 cm in diameter and invaded the eyelid margins and the posterior lamella of the eyelids. Immunohistochemistry labeling showed strong positivity for approximately 98.5% of S100 and positive staining for approximately 27.6% of Ki-67, 72.4% of Melan A and 6.8% of HMB45. However, Ki-67 was significantly upregulated in verrucous divided nevus of the eyelids compared with non-verrucous divided nevus, with approximately 38.8% upregulation in verrucous and 18.3% upregulation in non-verrucous nevus. Conclusions This study correlated the clinic-pathologic features of verrucous divided eyelid nevus by means of statistically analyzing the varied clinical features and pathological impressions. The significant over-expression of S100 may be used as an indicator of divided nevus of the eyelids, and the over-expressed Ki-67 may contribute to the recurrence of verrucous divided nevus. High expression of HMB45 and Melan A may represent malignant transformation.

Metastasis and chemical resistance are the most serious problems in the treatment of highly aggressive uveal melanoma (UM). The newly identified lncRNA OUM1 is overexpressed in UM, functions as a catalyst and regulates protein tyrosine phosphatase (PTP) activity by binding to PTP receptor type Z1 (PTPRZ1), which plays an important role in cell proliferation, metastasis and chemotherapy resistance in the UM microenvironment. Hence, siRNAs that selectively knocking down the lncRNA OUM1 (siOUM1) and its target gene PTPRZ1 (siPTPRZ1) were designed to inhibit the OUM1/PTPRZ1 pathway to reduce PTP activity, and this reduction in activity interrupts protein tyrosine phosphorylation, suppresses UM proliferation and metastasis and improves cisplatin sensitivity in UM cells. Then, to overcome the limitations of the difficulty of drug administration and traditional therapeutics, the indocyanine green (ICG)-labeled manganese metal-organic framework (MOF) nanoparticles (NPs) were fabricated and linked with arginine-glycine-aspartate (RGD) peptide to carry siOUM1/siPTPRZ1 and cisplatin to achieve targeted siRNA interference-mediated therapy, enhanced cisplatin therapy and chemodynamic therapy. This NP system also has a dual-modal imaging ability because ICG is a near-infrared region fluorescent dye and manganese has the potential to be used in magnetic resonance imaging. This study verifies the significance of the newly discovered lncRNA OUM1 as a new therapeutic target for aggressive UM and provides a drug delivery NP system for precise treatment of UM accompanied with a dual-modal imaging ability.

This study was designed to identify differently expressed circular RNAs (circRNAs) and investigate their potential roles in lacrimal sacs from patients with chronic dacryocystitis. The lacrimal sac samples of three chronic dacryocystitis patients and three control subjects were collected for RNA sequencing after ribosomal RNA was depleted. Differently expressed circRNAs and messenger RNAs (mRNAs) were used for co-expression analysis. CircRNA-microRNA (miRNA)-mRNA interaction network were also established by miRanda software. Meanwhile, pathway and functional enrichment analysis were conducted for the down- and up-regulated mRNAs in the circRNA-mRNA co-expression network. The expression levels of circRNAs and mRNAs in chronic dacryocystitis and control samples were validated by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). In all the 3,909 circRNAs predicted through RNA sequencing, 25 circRNAs (20 up-regulated and 5 down-regulated) expressed differently in chronic dacryocystitis samples. Besides, there identified 1,486 differentially expressed mRNAs. Of these differently expressed circRNAs and mRNAs, eight were validated by qRT-PCR, including MYH2, DSP, CD27, CCL5, FN1, has_circ_0004792, has_circ_0001062, and has_circ_0115476. Gene Ontology (GO) analysis indicated that the majority of altered mRNAs in this co-expression network were involved in immune system processes and meanwhile Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that these altered expressed mRNAs were also amplified in bacterial invasion of epithelial cells, both of which were thought to be involved in the pathogenesis of chronic dacryocystitis. In the circRNA-miRNA-mRNA interaction network, six circRNAs were found to be related to Th1 and Th2 cell differentiation, which was closely associated with the development of chronic dacryocystitis. This study identified statistically significant differences between circRNAs and mRNAs of lacrimal sac samples of chronic dacryocystitis patients and control individuals and provides novel insight into the regulatory mechanism of circRNAs, miRNAs, and mRNAs in the pathogenesis of chronic dacryocystitis.

Background Primary canaliculitis is a chronic infection of the proximal lacrimal pathway. We aimed to evaluate surgical outcomes of a canaliculoplasty procedure for primary canaliculitis associated with canalicular dilatation. Methods This study enrolled 42 primary canaliculitis patients with canalicular dilatation who underwent canaliculoplasty. All patients were treated with canaliculotomy, curettage of canalicular contents and canaliculoplasty with stent placement. Patients’ demographics, clinical features, and follow-up outcomes were evaluated. Results There were 12 males and 30 females with a mean age of 66.1 ± 13.9 years. The mean duration time from the first onset of signs/symptoms to diagnosis was 30.6 ± 39.5 months. Epiphora (90.5%) and mucopurulent discharge from punctum (85.7%) were the most common signs. Thirty-three out of 42 patients (78.6%) achieved complete remission with a mean follow-up time of 25.3 ± 12.9 months. There were 3 patients found to have canalicular stenosis due to obstruction after surgery. Conclusion Canalicular dilatation is a severe condition of primary canaliculitis, probably due to a combined result of long standing disease and the presence of concretions. The surgical procedure of canaliculoplasty can be a highly effective treatment for primary canaliculitis associated with canalicular dilatation.

Uveal melanoma (UM) is the most common primary intraocular carcinoma in adults. Cinobufagin, secreted by the Asiatic toad Bufo gargarizans, is a traditional Chinese medicine, widely used in tumor treatment. Here, we explored the potential antitumor function of cinobufagin and investigated its biochemical mechanisms in UM cells. The antitumor potential of cinobufagin was determined via cell viability, cell cycle, and apoptosis assays. Colony formation assays confirmed that cinobufagin exerted potent antitumor activity in a dose-dependent manner. We found that cinobufagin could induce cell apoptosis and upregulate the expression of cleaved caspase-3, cleaved poly(ADP-ribose) polymerase (PARP), and cleaved caspase-9 in vivo and in vitro. In addition, after treatment with increased concentrations of cinobufagin, the intrinsic mitochondrial apoptosis pathway was also activated, which was demonstrated by increased cell apoptosis with increased expression of Bad and Bax, decreased expression of Bcl-2 and Bcl-xl, and reduced mitochondrial membrane potential (MMP) in OCM1 cells. Taken together, the results of this preclinical study suggest that cinobufagin can both inhibit cell survival and induce cell apoptosis in a dose-dependent manner in UM cells, which provides new insights into the biochemical mechanism of cinobufagin and its potential as a future chemotherapeutic agent for UM.

Purpose: To report a case with neurofibromatosis type 1 presenting as prominent enophthalmos and abnormal infraorbital artery. Case description: A 19-year-old man with a family history of neurofibromatosis presented with prominent right enophthalmos. Computed tomography showed orbital dysplasia and enlarged inferior orbital fissure but no plexiform neurofibroma. Prominent intraoperative hemorrhage originated from several abnormal arteries in the infraorbital region during orbital reconstruction. A tortuous and dysplastic infraorbital artery was verified postoperatively by computed tomography angiography. The bleeding vessels were supposed to be the orbital branches of the dysplastic infraorbital artery. Conclusions: The orbital malformation and enlargement of inferior orbital fissure probably resulted in an abnormal infraorbital artery. Selective artery embolization may be chosen as a preceding treatment before orbital reconstruction surgery.

BACKGROUND:Long non-coding RNAs (lncRNAs) have been identified as important epigenetic regulators that play critical roles in human cancers. However, the regulatory functions of lncRNAs in tumorigenesis remains to be elucidated. Here, we aimed to investigate the molecular mechanisms and potential clinical application of a novel lncRNA, retinoblastoma associated transcript-1 (RBAT1), in tumorigenesis.METHODS:RBAT1 expression was determined by real-time PCR in both retinoblastoma (Rb) and bladder cancer (BCa) cell lines and clinical tissues. Chromatin isolation using RNA purification (ChIRP) assays were performed to identify RBAT1-interacting proteins. Patient-derived xenograft (PDX) retinoblastoma models were established to test the therapeutic potential of RBAT1-targeting GapmeRs.RESULTS:Here, we found that RBAT1 expression was significantly higher in Rb and BCa tissues than that in adjacent tissues. Functional assays revealed that RBAT1 accelerated tumorigenesis both in vitro and in vivo. Mechanistically, RBAT1 recruited HNRNPL protein to E2F3 promoter, thereby activating E2F3 transcription. Therapeutically, GapmeR-mediated RBAT1 silencing significantly inhibited tumorigenesis in orthotopic xenograft retinoblastoma models derived from Rb cell lines and Rb primary cells.CONCLUSIONS:RBAT1 overexpression upregulates a known oncogene, E2F3, via directly recruiting HNPNPL to its promoter and cis-activating its expression. Our finding provides a novel mechanism of lncRNA biology and provides potential targets for diagnosis and treatment of Rb and BCa.

BACKGROUND:To introduce a modified deep anterior lamellar dissection technique to improve visibility during surgery for vitreoretinal diseases with coexisting corneal opacity.CASE PRESENTATION:Two patients with retinal detachment and coexisting corneal blood staining or corneal decompensation underwent modified deep anterior lamellar dissections followed by vitrectomy. The modified deep anterior lamellar dissection techniques, unlike the dissection and removal of corneal lamellar in a typical deep anterior lamellar keratoplasty, included the creation and preservation of a deep lamellar corneal flap, the retroillumination to visualize and easily remove the remaining opaque stroma on the Descemet membrane, and the big air bubble technique in the eye with endothelial decompensation. The patient's own cornea flap was sutured back after vitrectomy was done. The modified dissection techniques provided adequate fundus view during vitrectomy while removing as less corneal tissue as possible and decreasing the surgical complications and the requirement of a fresh cornea. Postoperatively, in case 1, the corneal blood staining was gradually absorbed and the vision improved from light perception to counting fingers. In case 2, even though the cornea remained cloudy and the vision was poor, the cornea endothelial decompensation was stable and asymptomatic. Both retinas were attached after silicone oil removal at 6-month follow-up.CONCLUSIONS:This modified and limited deep anterior lamellar corneal dissection procedure appears to be a useful alternative to penetrating keratoplasty, ophthalmic endoscope and temporary keratoprosthesis during the vitrectomy with coexisting corneal opacity.

The NADPH oxidase Nox4 is a multi-pass membrane protein responsible for the generation of reactive oxygen species that are implicated in cellular signaling but may also cause pathological situations when dysregulated. Although topological organization of integral membrane protein dictates its function, only limited experimental data describing Nox4's topology are available. To provide deeper insight on Nox4 structural organization, we developed a novel method to determinate membrane protein topology in their cellular environment, named Topological Determination by Ubiquitin Fusion Assay (ToDUFA). It is based on the proteolytic capacity of the deubiquitinase enzymes to process ubiquitin fusion proteins. This straightforward method, validated on two well-known protein's topologies (IL1RI and Nox2), allowed us to discriminate rapidly the topological orientation of protein's domains facing either the nucleocytosolic or the exterior/luminal compartments. Using this method, we were able for the first time to determine experimentally the topology of Nox4 which consists of 6 transmembrane domains with its N- and C-terminus moieties facing the cytosol. While the first, third and fifth loops of Nox4 protein are extracellular; the second and fourth loops are located in the cytosolic side. This approach can be easily extended to characterize the topology of all others members of the NADPH oxidase family or any multi-pass membrane proteins. Considering the importance of protein topology knowledge in cell biology research and pharmacological development, we believe that this novel method will represent a widely useful technique to easily uncover complex membrane protein's topology.

Background: MicroRNAs (miRNAs) have been shown to function in many different cellular processes, including proliferation, apoptosis, differentiation and development. miR-181a, -181b, -181c and -181d are miR-181 members of the family, which has been rarely studied, especially uveal melanoma. Methods: The expression level of miR-181 family in human uveal melanoma cell lines was measured via real-time PCR (RT-PCR). The function of miR-181 on cell cycle was detected through Flow Cytometry assay. Microarray assay and Bioinformatics analysis were used to find the potential target of miR-181b, and dual-luciferase reporter assays further identified the target gene. Results: MiR-181 family members were found to be highly homologous across different species and their upregulation significantly induces UM cell cycle progression. Of the family members, miR-181b was significantly overexpressed in UM tissues and most UM cells. Bioinformatics and dual luciferase reporter assay confirmed CTDSPL as a target of miR-181b. miR-181b over-expression inhibited CTDSPL expression, which in turn led to the phosphorylation of RB and an accumulation of the downstream cell cycle effector E2F1, promoting cell cycle progression in UM cells. Knockdown CTDSPL using siRNAs showing the same effect, including increase of E2F1 and the progression of cell cycle. Conclusions: MiR-181 family members are key negative regulators of CTDSPL-mediated cell cycle progression. These results highlight that miR-181 family members, especially miR-181b, may be useful in the development of miRNA-based therapies and may serve as novel diagnostic and therapeutic candidate for UM.

Increasing evidence suggests that aberrant long non-coding RNAs (lncRNAs) are significantly correlated with the pathogenesis, development and metastasis of cancers. RHPN1 antisense RNA 1 (RHPN1-AS1) is a 2030-bp transcript originating from human chromosome 8q24. However, the role of RHPN1-AS1 in uveal melanoma (UM) remains to be clarified. In this study, we aimed to elucidate the molecular function of RHPN1-AS1 in UM. The RNA levels of RHPN1-AS1 in UM cell lines were examined using the quantitative real-time polymerase chain reaction (qRT-PCR). Short interfering RNAs (siRNAs) were designed to quench RHPN1-AS1 expression, and UM cells stably expressing short hairpin (sh) RHPN1-AS1 were established. Next, the cell proliferation and migration abilities were determined using a colony formation assay and a transwell migration/invasion assay. A tumor xenograft model in nude mice was established to confirm the function of RHPN1-AS1 in vivo. RHPN1-AS1 was significantly upregulated in a number of UM cell lines compared with the normal human retinal pigment epithelium (RPE) cell line. RHPN1-AS1 knockdown significantly inhibited UM cell proliferation and migration in vitro and in vivo. Our data suggest that RHPN1-AS1 could be an oncoRNA in UM, which may serve as a candidate prognostic biomarker and target for new therapies in malignant UM.