The Role Of Lncrna RHPN1-AS1 In UM Tumorigenesis And Chol-siRNA SDF-1 In Angiogenesis Inhibition

Purpose: Increasing evidences suggest that aberrant lnc RNAs are significantly correlated with the pathogenesis,development and metastasis of cancers.RHPN1 antisense RNA 1(RHPN1-AS1)is a 2013-bp transcript on human chromosome 8q24.However,the role of RHPN1-AS1 in uveal melanoma remains to be clarified.In this study,we aimed to elucidate the molecular functions of RHPN1-AS1 in UM.Methods: The RNA levels of RHPN1-AS1 in UM cell lines were examined by quantitative real-time polymerase chain reaction(q RT-PCR).Si RNAs were designed to silence RHPN1-AS1 expression and UM cells stably expressed sh RHPN1-AS1 were established.Next,Cell proliferation and migration ability were determined by the colony formation assay and transwell migration assay.Finally,the tumor xenograft model in nude mice was established to confirm the function of RHPN1-AS1 in vivo after transplanted with sh RHPN1-AS1-OCM1 and untreated OCM1 cells.Results: RHPN1-AS1 was significantly upregulated in a number of UM cell lines compared with normal human retinalpigment epithelium(RPE)cell line.RHPN1-AS1 knockdown could significantly inhibit UM cells proliferation and migration in vitro and in vivo.Conclusions: Our data suggest that RHPN1-AS1 could be an onco RNA in UM,which may served as a candidate prognostic biomarker and target for new therapies in malignant UM.Purpose.Ophthalmologist is not willing to see neovascularization(NV)during the ocular tissue repair.NV in optical axiswill cause of visual loss.Cholesterol modify of si RNA allow independent cell transfect without lipo-complex and slow down the RNA degration.SDF-1 chol-si RNA was designed to prevent the expression of chemokine SDF-1.Its angiogenesis effect in vitro and vivo were detected.Method: The ratendothelial progenitor cells(EPCs)and bone marrow mesenchymal stem cells(BMSCs)were separated and cultured in vitro.SDF-1 m RNA expression and the silence effect in vitro was detected by q PCR.Silence effectin vivo were detected by q PCR and ELISA afterintra-vitreous injection.Cell migration was detected in transwellsystem.Angiogenesis of SDF-1 chol-si RNA were test in corneal alkali burn.Result: Rat EPCs and BMSCs showed a high level of SDF-1 m RNA expression and the m RNA expression can be prevent by both normal si RNA and chol-si RNA.Cholesterol modified SDF-1 si RNA showed independent cell transfection ability in vitro and long silence effect in vivo.SDF-1 chol-si RNA inhibited BMSCs migration in vitro and showed angiogenesis effect in corneal alkali burn model.Conclusion: Cholesterol modify proved to be an excellent molecule modification of si RNA to provide anindependent cell transfection with longer silence period.SDF-1 chol-si RNA can effectively reduce the expression of SDF-1,prevent cell migration and angiogenesis effect in vitro and in vivo.Purpose: To compare choroidal microstructural changes in eyes with age-related macular degeneration(AMD)of different stages.Design: Retrospective,cross-sectional case series.Participants: Thirty-two age-matched normal eyes as controls,26 fellow uninvolved eyes of intermediate/late AMD,29 eyes of early AMD,28 eyes of intermediate AMD,and 39 eyes of late AMD were included in the study.Methods: All subjects underwent comprehensive ophthalmological examination.The choroid images,including subfoveal choroidal thickness,percentage of Sattler’s layer area,and en face images of the choroid were obtained using spectral-domain optical coherence tomography.Main Outcome Measures: Subfoveal choroidal thickness changes,percentage of Sattler’s layer area changes,and en-face images of the choroid in AMD eyes were measured.Results: One hundred fifty-four eyes of 96 individuals with mean age of 67.1±9.2 years were included.The mean subfoveal choroidal thickness was 295.4±56.8 μm in age-matched normal eyes,306.7±68.4 μm in fellow uninvolved eyes with AMD,293.8±80.4 μm in early AMD,215.6±80.4 μm in intermediate AMD,and 200.4±66.6 μm in late AMD(F=14.2,all P<0.001).Choroidal thickness was greater in early AMD eyes than in intermediate/late AMD eyes(P<0.001).Mean percentage of Sattler’s area in each group showed a similar tendency.The microstructure of the choroid showed reduced vascular density of Sattler’s areas in late AMD eyes compared to normal eyes.Conclusions: Decreasing subfoveal choroidal thicknesses and percentages of Sattler’s areas were found in the progression of AMD.The choroidal change was related to atrophy of the microstructural changes of underlying capillaries and medium-sized vessels.