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08.05.2019 | Original Article | Ausgabe 3/2019

Hepatology International 3/2019

C1QTNF1-AS1 regulates the occurrence and development of hepatocellular carcinoma by regulating miR-221-3p/SOCS3

Zeitschrift:
Hepatology International > Ausgabe 3/2019
Autoren:
Hang Li, Bo Zhang, Meng Ding, Shang Lu, Hui Zhou, Dajun Sun, Gang Wu, Xianfeng Gan
Wichtige Hinweise

Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1007/​s12072-019-09944-5) contains supplementary material, which is available to authorized users.

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Abstract

Background

The aim of our study was to explore how C1QTNF1-AS1 regulated miR-221-3p/SOCS3 axis in human hepatocellular carcinoma (HCC).

Methods

Differentially expressed lncRNAs and genes were examined via RNA-seq. GO analysis and KEGG pathway enrichment analysis were carried out based on the function of dys-regulated mRNAs. RT-qPCR was employed to detect the relative mRNA expression level of C1QTNF1-AS1, miR-221-3p, SOCS3 and key genes in the JAK/STAT signaling pathway in HCC tissues and cells, and western blot analysis was conducted to detect the relative protein expression levels of SOCS3 and key proteins in the JAK/STAT signaling pathway in HCC tissues and cells. MTT assay, transwell assay and flow cytometry were utilized to assess HCC cell proliferation, invasion, migration and apoptosis. Dual luciferase reporter gene assay was used to verify the targeted relationship between C1QTNF1-AS1 and miR-221-3p, as well as between miR-221-3p and SOCS3. A tumorigenicity assay in nude mice was conducted to investigate the effects of C1QTNF1-AS1 on HCC tumor growth in vivo.

Results

C1QTNF1-AS1 and SOCS3 were down-regulated, while miR-221-3p was up-regulated in HCC tissues and cells. In HepG2 and Huh7 cells, overexpression of C1QTNF1-AS1 or SOCS3, as well as silence of miR-221-3p inhibited HCC cell proliferation, migration, and invasion and promoted HCC cell apoptosis. The results of the dual luciferase reporter gene assay indicated that miR-221-3p could directly target both C1QTNF1-AS1 and SOCS3. In addition, up-regulation of C1QTNF1-AS1 suppressed HCC tumor growth in vivo.

Conclusion

Overexpression of C1QTNF1-AS1 down-regulated miR-221-3p and subsequently up-regulated SOCS3, thereby inhibiting HCC cell proliferation, migration and invasion and promoting apoptosis through the JAK/STAT signaling pathway.

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Zusatzmaterial
Supplementary file1 (DOCX 16 kb)
12072_2019_9944_MOESM1_ESM.docx
Supplementary file2 (DOCX 15 kb)
12072_2019_9944_MOESM2_ESM.docx
Supplementary file3 (DOCX 15 kb)
12072_2019_9944_MOESM3_ESM.docx
Supplementary Figure 1 RNA-seq analysis defined down-regulation of C1QTNF1-AS1 and SOCS3 in HCC tissues. (A) (B) Mass fraction diagram of sample base sequences; (C) Heatmap of the ten most up-regulated and down-regulated mRNAs in normal and tumor tissues, in which SOCS3 was included; (D) Heatmap of the ten most up-regulated and down-regulated lncRNAs in normal and tumor tissues, in which C1QTNF1-AS1 was included (TIF 22440 kb)
12072_2019_9944_MOESM4_ESM.tif
Supplementary Figure 2 GO function enrichment and pathway enrichment analysis. (A)(B)(C) GO function analysis cluster diagram. A higher z-score represents an increase in function expression; (D) KEGG analysis revealed the different biological functions in cancer and normal samples; (E) Gene enrichment in the JAK/STAT pathway (TIF 33324 kb)
12072_2019_9944_MOESM5_ESM.tif
Supplementary Figure 3 (A) Effects of C1QTNF1-AS1 on apoptosis analysis of HCC cells; (B) Effects of C1QTNF1-AS1/miR-221-3p on apoptosis analysis of HCC cells; (C) Effects of SOCS3/miR-221-3p on apoptosis analysis of HCC cells (TIF 14187 kb)
12072_2019_9944_MOESM6_ESM.tif
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