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OncoTargets and Therapy Volume 11, 2018 - Issue
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Original Research

lncRNA FBXL19-AS1 regulates osteosarcoma cell proliferation, migration and invasion by sponging miR-346

Runsang Pan1 Department of Clinical Medicine, Guizhou University, Guiyang, Guizhou Province, China, [email protected]; [email protected];2 Department of Orthopedics, Guiyang Children’s Hospital, Guiyang, Guizhou Province, China
,
Zhixu He2 Department of Orthopedics, Guiyang Children’s Hospital, Guiyang, Guizhou Province, China
,
Wanyuan Ruan2 Department of Orthopedics, Guiyang Children’s Hospital, Guiyang, Guizhou Province, China
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Sun Li1 Department of Clinical Medicine, Guizhou University, Guiyang, Guizhou Province, China, [email protected]; [email protected];3 Department of Orthopedics, Guizhou Provincial People’s Hospital, Guiyang, Guizhou Province, China, [email protected]; [email protected]
,
Hui Chen1 Department of Clinical Medicine, Guizhou University, Guiyang, Guizhou Province, China, [email protected]; [email protected];3 Department of Orthopedics, Guizhou Provincial People’s Hospital, Guiyang, Guizhou Province, China, [email protected]; [email protected]
,
Zhiyu Chen1 Department of Clinical Medicine, Guizhou University, Guiyang, Guizhou Province, China, [email protected]; [email protected];3 Department of Orthopedics, Guizhou Provincial People’s Hospital, Guiyang, Guizhou Province, China, [email protected]; [email protected]
,
Fang Liu1 Department of Clinical Medicine, Guizhou University, Guiyang, Guizhou Province, China, [email protected]; [email protected]
,
Xiaobin Tian1 Department of Clinical Medicine, Guizhou University, Guiyang, Guizhou Province, China, [email protected]; [email protected];3 Department of Orthopedics, Guizhou Provincial People’s Hospital, Guiyang, Guizhou Province, China, [email protected]; [email protected]Correspondence[email protected] [email protected]
&
Yingjie Nie1 Department of Clinical Medicine, Guizhou University, Guiyang, Guizhou Province, China, [email protected]; [email protected];3 Department of Orthopedics, Guizhou Provincial People’s Hospital, Guiyang, Guizhou Province, China, [email protected]; [email protected]Correspondence[email protected] [email protected]
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Pages 8409-8420 | Published online: 29 Nov 2018

Figures & data

Figure 1 lncRNA FBXL19-AS1 was upregulated in OS tissues and cell lines.

Notes: (A) FBXL19-AS1 was significantly upregulated in OS tissues compared with normal tissues. (B) FBXL19-AS1 was significantly upregulated in OS cell lines compared with hFOB1.19 cells. Stable FBXL19-AS1 knockdown in (C) MG63 and (D) 143B cells was confirmed by RT-PCR. *P<0.05.
Abbreviations: FBXL19-AS1, FBXL19 antisense RNA 1; OS, osteosarcoma; RT-PCR, reverse transcriptase PCR.
Figure 1 lncRNA FBXL19-AS1 was upregulated in OS tissues and cell lines.

Figure 2 lncRNA FBXL19-AS1 promoted OS cell proliferation in vitro.

Notes: (A) FBXL19-AS1 knockdown suppressed OS cell proliferation, as measured by the CCK-8 assay. (B–C) Colony formation assays showed a significantly lower proliferative rate in both cell lines following FBXL19-AS1 knockdown. (D–E) Flow cytometry was used to explore whether FBXL19-AS1 promoted OS cell proliferation by regulating cell cycle progression. (F–G) Western blot analysis showing FBXL19-AS1 knockdown downregulated cyclin D1, CDK2, and CDK4, which promote cell cycle progression. *P<0.05.
Abbreviations: FBXL19-AS1, FBXL19 antisense RNA 1; OS, osteosarcoma.
Figure 2 lncRNA FBXL19-AS1 promoted OS cell proliferation in vitro.

Figure 3 lncRNA FBXL19-AS1 promoted OS cell tumor growth in vivo.

Notes: (A) The FBXL19-AS1 knockdown group showed a reduced tumor growth rate. (B–C) The FBXL19-AS1 knockdown group showed reduced tumor diameters and volumes compared with controls, and the tumor-bearing mice lost less weight. *P<0.05; **P<0.01.
Abbreviations: FBXL19-AS1, FBXL19 antisense RNA 1; OS, osteosarcoma.
Figure 3 lncRNA FBXL19-AS1 promoted OS cell tumor growth in vivo.

Figure 4 lncRNA FBXL19-AS1 promoted OS cell migration and invasion in vitro.

Notes: (A–D) Wound healing assays showed significantly decreased migratory abilities in both cell lines following FBXL19-AS1 knockdown. (E–F) Transwell assays showed significantly decreased migration abilities in both cell lines following FBXL19-AS1 knockdown. (G–H) Transwell assays showed significantly decreased invasive abilities in both cell lines following FBXL19-AS1 knockdown. *P<0.05.
Abbreviations: FBXL19-AS1, FBXL19 antisense RNA 1; h, hours; OS, osteosarcoma.
Figure 4 lncRNA FBXL19-AS1 promoted OS cell migration and invasion in vitro.

Figure 5 miR-346 is a FBXL19-AS1 target.

Notes: (A) We found that miR-346 could potentially bind FBXL19-AS1 using starBASE 2.0. (B) FISH staining showed the localization of FBXL19-AS1 in MG63 and 143B cells. (C, D) RT-PCR results indicated FBXL19-AS1 was primarily expressed in the cytosol; GAPDH and U6 served as cytoplasmic and nuclear controls, respectively. (E) qPCR showed that FBXL19-AS1 knockdown upregulated miR-346 expression in MG63 and 143B cells. (F) RT-PCR showed that miR-346 expression was markedly reduced in OS cell lines. (G) RT-PCR showed that expression miR-346 was markedly reduced in OS tissues (P<0.01). (H) The correlation between FBXL19-AS1 and miR-346 expression in OS cell lines. (I) The correlation between FBXL19-AS1 and miR-346 expression in OS tissues. (J) miR-346 overexpression reduced the relative luciferase activity of pmirGLO-FBXL19-AS1-WT, while there was no significant reduction in the luciferase activity of pmirGLO-FBXL19-AS1-MUT. *P<0.05.
Abbreviations: FBXL19-AS1, FBXL19 antisense RNA 1; FISH, fluorescence in situ hybridization; MUT, mutation; NC, negative control; NS, not significant; OS, osteosarcoma; RT-PCR, reverse transcriptase PCR.
Figure 5 miR-346 is a FBXL19-AS1 target.
Figure 5 miR-346 is a FBXL19-AS1 target.

Figure 6 miR-346 mediates the pro-proliferation roles of FBXL19-AS1 in OS.

Notes: (A) Cells co-transfected with anti-miR-346 rescued the inhibitory effect of sh-FBXL19-AS1 in the CCK-8 assay. (B, C) Cells co-transfected with anti-miR-346 rescued the inhibitory effect of sh-FBXL19-AS1 on colony formation. *P<0.05.
Abbreviations: FBXL19-AS1, FBXL19 antisense RNA 1; OS, osteosarcoma.
Figure 6 miR-346 mediates the pro-proliferation roles of FBXL19-AS1 in OS.

Figure 7 miR-346 mediates the pro-proliferative roles of FBXL19-AS1 in OS.

Notes: (A–D) Wound healing assays showed a reduced migratory ability in both cell lines following co-transfection of anti-miR-346 and sh-FBXL19-AS1. (E–H) Transwell assays indicated that miR-346 inhibition promoted the migration and invasion of sh-FBXL19-AS MG63 and 143B cells. Magnification 400×. *P<0.05; **P<0.01.
Abbreviations: FBXL19-AS1, FBXL19 antisense RNA 1; OS, osteosarcoma.
Figure 7 miR-346 mediates the pro-proliferative roles of FBXL19-AS1 in OS.

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