Data_Sheet_1_LncRNA RP11-499E18.1 Inhibits Proliferation, Migration, and Epithelial–Mesenchymal Transition Process of Ovarian Cancer Cells by Dissociating PAK2–SOX2 Interaction.XLSX

Background: Ovarian cancer (OC)is a deadly gynecological malignancy worldwide. It is urgent to identify diagnostic biomarkers of OC to disclose the underlying mechanism.Methods and Materials: Bioinformatics analysis was used to identify target genes. Gene expression was detected and altered by qRT-PCR and cell transfection, respectively. The interaction between RP11-499E18.1 and PAK2, as well as that between PAK2 and SOX2, was determined using RNA pulldown, RNA immunoprecipitation (RIP), and co-immunoprecipitation (co-IP) assay, respectively. Localizations of RP11-499E18.1, PAK2, and SOX2 were respectively determined employing immunohistochemical (IHC), IF, and FISH. The regulatory effects of RP11-499E18.1, PAK2, and SOX2 on OC cell proliferation, migration, colony formation, epithelial–mesenchymal transition (EMT)-related factor expression, and SOX2 nuclear translocation were determined. Finally, the effects of RP11-499E18.1 and PAK2 expression on the tumor growth in nude mice were determined.Results: RP11-499E18.1, PAK2, and SOX2 were selected in our study. RP11-499E18.1 was downregulated, while PAK2 and SOX2 was upregulated in OC tissues and cells. RP11-499E18.1 coexists in the nucleus and cytoplasm of OC cells. There is an interaction between RP11-499E18.1 and PAK2, as well as PAK2 and SOX2 in OC cells. Alteration of RP11-499E18.1 and PAK2 expression both had no influence on PAK2 and SOX2 levels, but PAK2 upregulation notably augmented p-SOX2 level. RP11-499E18.1 overexpression suppressed OC cell proliferation, migration, and colony formation, as well as SOX2 nuclear translocation. Besides, it inhibited tumor growth in nude mice. However, these effects were notably reversed by PAK2 upregulation and eventually offset by SOX2 knockdown. Additionally, RP11-499E18.1 overexpression reduced PAK2–SOX2 interaction and SOX phosphorylation, and increased the binding of RP11-499E18.1 by PAK2.Conclusion: These lines of evidence demonstrated that RP11-499E18.1 might play its tumor suppressor roles in OC via regulation of the RP11-499E18.1–PAK2–SOX2 axis. This research indicated that RP11-499E18.1 might be used as a diagnostic biomarker for OC in the future.</p

Data_Sheet_2_LncRNA RP11-499E18.1 Inhibits Proliferation, Migration, and Epithelial–Mesenchymal Transition Process of Ovarian Cancer Cells by Dissociating PAK2–SOX2 Interaction.DOCX

Background: Ovarian cancer (OC)is a deadly gynecological malignancy worldwide. It is urgent to identify diagnostic biomarkers of OC to disclose the underlying mechanism.Methods and Materials: Bioinformatics analysis was used to identify target genes. Gene expression was detected and altered by qRT-PCR and cell transfection, respectively. The interaction between RP11-499E18.1 and PAK2, as well as that between PAK2 and SOX2, was determined using RNA pulldown, RNA immunoprecipitation (RIP), and co-immunoprecipitation (co-IP) assay, respectively. Localizations of RP11-499E18.1, PAK2, and SOX2 were respectively determined employing immunohistochemical (IHC), IF, and FISH. The regulatory effects of RP11-499E18.1, PAK2, and SOX2 on OC cell proliferation, migration, colony formation, epithelial–mesenchymal transition (EMT)-related factor expression, and SOX2 nuclear translocation were determined. Finally, the effects of RP11-499E18.1 and PAK2 expression on the tumor growth in nude mice were determined.Results: RP11-499E18.1, PAK2, and SOX2 were selected in our study. RP11-499E18.1 was downregulated, while PAK2 and SOX2 was upregulated in OC tissues and cells. RP11-499E18.1 coexists in the nucleus and cytoplasm of OC cells. There is an interaction between RP11-499E18.1 and PAK2, as well as PAK2 and SOX2 in OC cells. Alteration of RP11-499E18.1 and PAK2 expression both had no influence on PAK2 and SOX2 levels, but PAK2 upregulation notably augmented p-SOX2 level. RP11-499E18.1 overexpression suppressed OC cell proliferation, migration, and colony formation, as well as SOX2 nuclear translocation. Besides, it inhibited tumor growth in nude mice. However, these effects were notably reversed by PAK2 upregulation and eventually offset by SOX2 knockdown. Additionally, RP11-499E18.1 overexpression reduced PAK2–SOX2 interaction and SOX phosphorylation, and increased the binding of RP11-499E18.1 by PAK2.Conclusion: These lines of evidence demonstrated that RP11-499E18.1 might play its tumor suppressor roles in OC via regulation of the RP11-499E18.1–PAK2–SOX2 axis. This research indicated that RP11-499E18.1 might be used as a diagnostic biomarker for OC in the future.</p

Images under SEM and EDX traces of the scaffold with 2.5% ZnO doped after SBF incubation for 7 days.

<p>Images under SEM and EDX traces of the scaffold with 2.5% ZnO doped after SBF incubation for 7 days.</p

The morphology of initial MG-63 cells visualized by light microscopy (a), the morphology of MG-63 cells cultured on the scaffolds after 5 days by SEM (b) pure TCP; (c) 0.5 wt% ZnO; (d) 1.5 wt% ZnO; (e) 2.5 wt% ZnO and (f) 3.5 wt% ZnO, the fluorescence microscope images of MG-63 cells before (g) or after (h) incubation on the scaffolds with 2.5 wt% ZnO doping.

<p>The morphology of initial MG-63 cells visualized by light microscopy (a), the morphology of MG-63 cells cultured on the scaffolds after 5 days by SEM (b) pure TCP; (c) 0.5 wt% ZnO; (d) 1.5 wt% ZnO; (e) 2.5 wt% ZnO and (f) 3.5 wt% ZnO, the fluorescence microscope images of MG-63 cells before (g) or after (h) incubation on the scaffolds with 2.5 wt% ZnO doping.</p

Representative porous TCP scaffold under compression test (a); overview of scaffold after compression test (b); Effect of ZnO contents on the compressive strength (c) and stiffness (d) of porous scaffold; Effect of ZnO contents on the densification (e), microhardness (f) and fracture toughness (g) of the strut.

<p>Data represents mean ± standard deviation for <i>n = </i>6, *<i>P</i><0.05 and <i>**P<</i>0.01 (compared with pure TCP scaffold).</p

Micrographs of (a) raw β-TCP powder; (b) raw ZnO powder; (c) unetched surface of scaffold; etched surface of scaffold: (d) pure TCP; (e) 0.5 wt% ZnO; (f) 1.5 wt% ZnO; (g) 2.5 wt% ZnO and (h) 3.5 wt% ZnO.

<p>Micrographs of (a) raw β-TCP powder; (b) raw ZnO powder; (c) unetched surface of scaffold; etched surface of scaffold: (d) pure TCP; (e) 0.5 wt% ZnO; (f) 1.5 wt% ZnO; (g) 2.5 wt% ZnO and (h) 3.5 wt% ZnO.</p

XRD patterns (A): 2θ from 10 to 40° and (B): 2θ from 30.9 to 31.15° of (a) the raw β-TCP powder, the scaffolds with different ratio of ZnO: (b) 0 wt%; (c) 0.5 wt%; (d) 1.5 wt%; (e) 2.5 wt% and (f) 3.5 wt%. (β-TCP, JCPDS file number 09-0169; α-TCP, JCPDS file number 09-0348; ZnO, JCPDS file number 36-1451).

<p>XRD patterns (A): 2θ from 10 to 40° and (B): 2θ from 30.9 to 31.15° of (a) the raw β-TCP powder, the scaffolds with different ratio of ZnO: (b) 0 wt%; (c) 0.5 wt%; (d) 1.5 wt%; (e) 2.5 wt% and (f) 3.5 wt%. (β-TCP, JCPDS file number 09-0169; α-TCP, JCPDS file number 09-0348; ZnO, JCPDS file number 36-1451).</p

Weight loss assay of the scaffolds after incubation with SBF for different days.

<p>Data represents mean ± standard deviation for <i>n = </i>6, *<i>P</i><0.05 (compared to pure TCP scaffold).</p

Scaffold model designed using SolidWorks software: D = 1.2 mm; H = 11 mm; L = 18 mm.

<p>Scaffold model designed using SolidWorks software: D = 1.2 mm; H = 11 mm; L = 18 mm.</p

Summary of the properties of human bone.

<p>Summary of the properties of human bone.</p