Long Noncoding RNA Expression Profile In HLE B-3 Cells During TGF-β2-induced Epithelial-mesenchymal Transition

Objective:Recent evidence has shown that long noncoding RNAs（lnc RNAs）are involved in the process of epithelial-mesenchymal transition（EMT）.However,little research has focused on the expression profile of lnc RNAs during EMT in human lens epithelial cells（LECs）.We predicted lnc RNAs were involved in the process of EMT in LECs.We designed this study using microarray to find differentially expressed lnc RNAs in LECs which were during EMT,and could introduce new avenues for the prevention and treatment of posterior capsule opacification（PCO）.Methods:LECs were induced by TGF-β₂for 24 h.Then we used inverted microscope to observe cell morphology change.We confirmed EMT relative biomarkers including E-cadherin、α-SMA and vimentin via immunofluorescence,quantitative real-time reverse transcription polymerase chain reaction（q RT-PCR）,and western blot.Dysregulated lnc RNAs and m RNAs in normal human lens epithelial B-3（HLE B-3）cells and during TGF-β₂-induced EMT were analyzed via lnc RNA microarray.Gene Ontology（GO）and KEGG Pathway analyses of differentially expressed m RNAs were performed to identify their functions and pathologic pathways.Six candidate lnc RNAs were validated via q RT-PCR to confirm the microarray data.Results:After induced by TGF-β₂,cells were transformed from single polygonal to long and spindle-shaped.The results of immunofluorescence,q RT-PCR,and western blot showed,compared to control group,E-cadherin expression decreased,whileα-SMA and vimentin expression increased.A total of 775 lnc RNAs（325 up-regulated and 450 down-regulated）and 935 m RNAs（329 up-regulated and 606 down-regulated）were differentially expressed in HLE B-3 cells during TGF-β₂-induced EMT compared to normal HLE B-3cells.GO and KEGG Pathway analyses indicated the functions of differentially expressed m RNAs in the TGF-β₂-induced EMT in HLE B-3 cells.QRT-PCR confirmed the trends indicated in microarray analysis for all 6 candidate lnc RNAs.Conclusions:Compared with normal control,lnc RNAs expression in TGF-β₂-induced LECs significantly changed,indicating that lnc RNAs were involved in the process of EMT in LECs.