LncRNA PVT1 knockdown affects proliferation and apoptosis of uveal melanoma cells by inhibiting EZH2.

OBJECTIVE: To detect the expression of long non-coding ribonucleic acid (lncRNA) plasmacytoma variant translocation gene 1 (PVT1) in uveal melanoma (UM) tissues, and to investigate its influence on the proliferation and apoptosis of UM cells as well as its mechanism. PATIENTS AND METHODS: 40 cases of UM tissues and 40 cases of adjacent tissues surgically resected in our hospital from October 2015 to April 2018 were collected. The expression level of lncRNA PVT1 in these tissues was determined by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Stable knockdown of lncRNA PVT1 was constructed in human UM cell line OCM-1 using small interfering RNA (siRNA). The impact of lncRNA PVT1 on UM cell proliferation was detected by Cell Counting kit-8 (CCK-8) and colony formation assay. Flow cytometry was applied to measure the apoptotic level of UM cells in the blank control group and lncRNA PVT1 knockdown group. Meanwhile, the expression level of enhancer of zeste homologue 2 (EZH2) was determined by Western blotting. RESULTS: The expression level of lncRNA PVT1 in UM tissues was remarkably higher than that in the adjacent tissues (p<0.05). UM cell proliferation was notably repressed after lncRNA PVT1 knockdown by siRNA. Flow cytometry results indicated that the number of apoptotic UM cells in lncRNA PVT1 knockdown group significantly increased compared with that in the blank control group (p<0.05). The protein expression of EZH2 was suppressed after lncRNA PVT1 knockdown (p<0.05). CONCLUSIONS: LncRNA PVT1 knockdown in UM cells can repress the proliferation of UM cells and promote their apoptosis by regulating EZH2 expression.

[Clinical analysis of orbital ectopic meningiomas].

Objective: To investigate the clinical features, imaging features, diagnoses and therapeutic regimens of orbital ectopic meningiomas. Methods: A retrospective study. All the clinical data of 10 patients (10 eyes) with orbital ectopic meningiomas were analyzed retrospectively from August 1999 to October 2016. They included imaging data such as color Doppler ultrasound, CT and MRI, and information of diagnosis, pathology, therapeutic regimen and prognosis. Results: The age of orbital ectopic meningiomas was 7 to 68 years old. There were 4 males and 6 females. The clinical manifestations were mainly with swollen eyelid (8 cases), exophthalmos (7 cases), visual impairment (2 cases) and accidentally found a mass in the orbit with physical examination(2 cases). The tumor was located in (5 cases) or outside (5 cases) the muscular funnel of the orbit. The imaging features of CT included the irregular shape, unclear border, and extraocular muscular adhesions (10 cases). T1 weighted image of MRI showed low and medium signals and T2 weighted image showed medium and high signals(8 cases). There was no definite diagnosis before surgery. Ten patients were treated with surgery, and it was confirmed intraoperatively that the tumors were not adjacent to the optic nerve and orbital periosteum. The pathological diagnoses were mostly epithelial meningiomas (9 cases). There were no significant changes in visual acuity, but temporary eye movement disorders occurred postoperatively (10 cases). The patients were followed for 6 months to 6 years. Two cases underwent recurrent at 3 months and 2 years after operation, and received treatment of γ-knife radiation therapy. The tumor reduced after γ-knife radiotherapy, and had no change in 2 years and 6 years, respectively. Conclusions: As a kind of rare orbital tumors, it is difficult to diagnose orbital ectopic meningiomas accurately. The clinical features and imaging findings can help to detect the disease, but lacking the characteristics of optic nerve sheath meningiomas and periosteal meningiomas. Surgical resection could achieve a good prognosis without visual impairment. If the tumor recurs after pathological diagnosis, γ-knife conformal radiation therapy may still be effective for tumor recurrence. (Chin J Ophthalmol, 2018, 54: 665-670).

[Clinical analysis of patients with orbital nonspecific inflammatory response diseases whose CT or MR images show enlargement of the infraorbital nerves].

Objective: To discuss the clinical features, imaging features, pathological patterns, treatment principles and prognosis of the orbital nonspecific inflammatory response diseases patients whose CT or MR images show enlargement of the infraorbital nerves. Methods: A retrospective case series study. Seven orbital disease patients who were treated at Tianjin Medical University Eye Hospital between March 2013 and May 2017 were included. All patients, imaging pictures showed enlargement of the infraorbital nerve. The medical histories, clinical featuers, imaging features, pathologies, serological examinations, therapeutic processes and prognosis were collected and analyzed. Results: The 7 patients included 4 males and 3 females aged from 55 to 68 years (the average age was 60). Bilateral involvement was present in 5 of 7 patients. The main clinical manifestations include proptosis, increasing of orbital pressure, impairment of visual functions, reduction in ocular motility, facial sensation, periocular ache, involvement of lymph node and salivary gland, etc. CT results showed enlargement of infraorbital nerve, accompanied with or without the destruction of bone. The inflammatory response may involve with extraocular muscles and lacrimal glands, or were shown as lesions with irregular shape and blurred borders. The MR images generally showed equal T(1) and equal T(2) signal, accompanied with evident enlargement of the infraorbital nerve. Of all the 7 patients, 6 underwent operation, and the pathology confirmed that 2 of the 6 were involved with inflammatory pseudotumors and the rest 4 were involved with IgG4-related ophthalmic disease (IgG4-ROD). The level of IgG4 in the serum were detected for 4 patients, the results of 3 were high and the other was normal. Of all the 7 patients,1 patient underwent conservative treatment, but disease recurred for several times, and the serum IgG4 level for the patient was higher than normal;1 inflammatory pseudotumor patient was cured completely only by surgery, and has a favourable prognosis; 1 IgG4-ROD patient accepted glucocorticoid, surgery, radiotherapy and chemotherapy, but disease recurred several times; The rest of the patients accepted glucocorticoid and operation, 2 were sensitive to glucocorticoid, 2 were not sensitive, and 1 involved with recurrence. Conclusions: If orbital nonspecific inflammation response was accompanied with enlargement of infraorbital nerve, it is suggested that patient is more likely to be involved with IgG4-ROD;Pathology and serological tests can be used for the diagnosis of IgG4-ROD, however treatment effect appears to be poor for most patients, and patients prone to relapse. (Chin J Ophthalmol, 2018, 54: 515-519).

[The value of DCE-MRI in assessing the course of thyroid associated ophthalmopathy].

Objective: To explore the feasibility of the semi-quantitative rectus extraocular muscle (EOM) parameters of dynamic contrast-enhanced magetic resonance imaging (DCE-MRI) in assessing the clinical course of thyroid associated ophthalmopathy (TAO). Methods: It was a retrospective case series study. A total of 136 cases of TAO were recruited from March 2011 to October 2012 in the Tianjin the first center hospital including 63 males and 72 females, aged 24.0-65.0 years, with an average age of (40.5±10.9) years. Forty healthy volunteers were recruited as control group (CG). According to clinical activity score (CAS), all TAO patients were divided into 2 groups, activity group (AG) and inactivity group (IAG). MRI and DCE-MRI orbit scan were performed in each subject. Drew time-intensity curves (TIC) by Siemens 3.0 MR (syngo) post-processing workstation. The semi-quantitative parameters of DCE-MRI were calculated. The semi-quantitative paramters based on TIC include early enhancement coefficient (EEC), maximum enhancement coefficient (Emax) and wash-out coefficient (WC(5min)). Kruskal-Wallis H rank test was used for comparing signal intensity among 3 groups, and Nemenyi test for pairwise comparison between groups. The DCE-MRI parameters (EEC, Emax, WC(5 min)) among groups were compared by one-way ANOVA, and Bonferroni t test is for pairwise comparison between groups. The diagnostic value of mean EEC, mean Emax, WC(5min) for assessment of the clinical course in TAO was analyzed by ROC curve. Results: There were significant difference in signal intensity (SI) of rectus EOM on T(2)WI among CG, AG and IAG, which is significantly different in 88 cases of AG including 45 cases of high intensity 51.1%, 23 cases of moderate intensity 26.1%, and 20 cases of low intensity 22.7%,compared with CG and IAG. EEC (P<0.05), Emax and WC5min values of rectus EOM of TAO group were significantly lower than those of CG(P<0.05), which values of rectus EOM of TAO active group of EEC are 0.63±0.06、0.61±0.05、0.56±0.09、0.57±0.09, and values of rectus EOM of TAO inactive group of EEC are 0.49±0.05、0.50±0.08、0.57±0.10、0.55±0.09. The values of rectus EOM of TAO active group of Emax are 1.35±0.09、1.28±0.09、1.21±0.17、1.25±0.10, and the values of rectus EOM of TAO inactive group of Emax are 1.04±0.06、1.05±0.10、1.20±0.19、1.16±0.11. The values of rectus EOM of TAO active group of WC(5 min) are 0.13±0.03、0.13±0.03、0.13±0.06、0.13±0.03 and the values of rectus EOM of TAO inactive group of WC5min are 0.08±0.02、0.79±0.03、0.11±0.06、0.09± 0.03. EEC (χ(2)=9.20, P<0.05), Emax and WC(5min) values of rectus EOM of TAO group were significantly lower than those of CG (P<0.05). EEC, Emax and WC(5min) values of medial rectus and inferior rectus EOM of IAG were significantly lower than those of AG(P<0.05). WC(5min) values of superior rectus EOM of IAG were significantly lower than those of AG (P<0.05). There were no differences in EEC and Emax values of lateral rectus and superior rectus EOM between IAG and AG (P>0.05). There were no differences in WC(5min) values of lateral rectus EOM between IAG and AG (P>0.05). The area under the curve (AUC) were 0.771, 0.879, 0.898 for mean EEC, mean Emax, and mean WC(5min), respectively. Conclusion: The semi-quantitative paramters of DCE-MRI can show the clinical activity of TAO patients and can be considered as the quantitative index of TAO activity staging. (Chin J Ophthalmol, 2017, 53: 430-435).

[Bone marrow-derived fibrocytes and thyroid-associated opthalmopathy].

Thyroid-associated opthalmopathy(TAO) is a common autoimmune syndrome affecting the thyroid and orbit, which can result in the fibrosis of extraoular muscles and hyperplasia of adipose tissue. Advanced TAO patients could even lose vision caused by cornea ulcer and congestion of optic nerve from expansion of the extraocular muscles and orbit fat. Currently, there are no therapies shown to prevent it, because its cellular and molecular mechanisms are not clear. Some studies have recently implicated bone marrow-derived fibroblast-like, called fibrocytes are involved in the pathogenesis of TAO. We reviewed and summarized the research advances of TAO and also the relationship between the fibrocytes and pathogenesis of TAO in the paper. (Chin J Ophthalmol, 2017, 53: 470-473).

SOST silencing promotes proliferation and invasion and reduces apoptosis of retinoblastoma cells by activating Wnt/ß-catenin signaling pathway.

This study aimed to investigate the effects of SOST and the Wnt/ß-catenin signaling pathway on the proliferation, migration, invasion, and apoptosis of human retinoblastoma cells. Fifty-five retinoblastoma and 21 normal retinal tissue samples were collected as the case group and control group, respectively. HXO-RB44 and SO-RB50 cells were selected and assigned into blank, negative control (NC), siRNA 1, siRNA 2, siRNA 3, IWR-1-endo 1, IWR-1-endo 2 and IWR-1-endo 3 groups. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to detect the expression of SOST, Wnt-1, and ß-catenin in the collected tissue samples. MTT assay, flow cytometry, transwell assay and the starch test were employed to determine the cell proliferation, cell cycle, apoptosis, invasion and migration after transfection. The qRT-PCR and western blotting were also used to detect the mRNA and protein expressions of SOST, Wnt-1, ß-catenin, C-myc, Cyclin D1, MMP-2 and MMP-9. The tumor formation in nude mice was conducted to evaluate the effects of SOST on the growth of a transplanted tumor. Compared with normal retinal tissues, the retinoblastoma tissues exhibited a downregulation of SOST but an upregulation of Wnt-1 and ß-catenin. The proliferation, invasion and migration of HXO-RB44 and SO-RB50 cells in the SOST-siRNA group were significantly higher than the cells in the blank and NC groups. The expressions of Wnt-1, ß-catenin, C-myc, Cyclin D1, MMP-2 and MMP-9 in the three SOST-siRNA groups were elevated, but the SOST decreased when compared with the blank and NC groups. SOST silencing promoted the growth of transplanted tumors in nude mice. These findings indicate that SOST silencing promotes the proliferation, invasion and migration, and decreases the apoptosis of human retinoblastoma cells by activating the Wnt/ß-catenin signaling pathway.

[Observations on 108 normal ophthalmic arteries in 80 patients by digital subtraction angiography].

By digital subtraction angiography, the authors made observations on 108 normal ophthalmic arteries in 80 patients with regard to the origins, courses, branching, morphology and anastomoses of the vessels. The merits and demerits of DSA in observing the ophthalmic artery were discussed.