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Long non-coding RNA XIST regulates PTEN expression by sponging miR-181a and promotes hepatocellular carcinoma progression

View Article: PubMed Central - PubMed

ABSTRACT

Background: Tumor metastasis often occurs in hepatocellular carcinoma (HCC) and influences the patient’s prognosis, and microRNAs are reported to play key roles in tumor metastasis. This study was conducted to explore the effect of microRNAs on HCC metastasis.

Methods: The levels of miR-181a in HCC tissues, adjacent tissues, metastatic HCC tissues, and non-metastatic HCC tissues at different stages were determined by qRT-PCR. Effect of miR-181a on the proliferation, invasion, and metastasis of HCC cells was estimated by cell counting kits-8 (CCK-8), wound-healing, and Transwell assays. Software analysis and luciferase assays were used to explore the target gene of miR-181a.

Results: MiR-181a was up-regulated in HCC tissues and its expression level in metastatic HCC tissues was much higher than in non-metastasis samples. PTEN was found to be a target gene of miR-181a. MiR-181a had multiple binding sites with the long non-coding RNA (lncRNA) XIST. The regulation of miR-181a on PTEN was mediated by lncRNA XIST. The proliferation and invasion of cells with siXIST were significantly enhanced compared with those of control cells, while knockdown of miR-181a abolished the enhancing effects.

Conclusions: MiR-181a can promote HCC metastasis by targeting PTEN, which is regulated by lncRNA XIST.

Electronic supplementary material: The online version of this article (doi:10.1186/s12885-017-3216-6) contains supplementary material, which is available to authorized users.

No MeSH data available.


Related in: MedlinePlus

MiR-181a promoted HCC cell invasion and EMT by activating AKT signaling. a The protein expression of EMT related genes (Snail, Slug, N-cadherin, Vimentin and E-cadherin), MMP-2, and MMP-9. b Western blot analysis of phosphorylated (active) AKT and mTOR in HCC cells transfected with miR-181a mimics or inhibitor. HCC, hepatocellular carcinoma. EMT, epithelial-to-mesenchymal transition. All values are mean ± SD. ** vs NC group, p < 0.01
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Fig4: MiR-181a promoted HCC cell invasion and EMT by activating AKT signaling. a The protein expression of EMT related genes (Snail, Slug, N-cadherin, Vimentin and E-cadherin), MMP-2, and MMP-9. b Western blot analysis of phosphorylated (active) AKT and mTOR in HCC cells transfected with miR-181a mimics or inhibitor. HCC, hepatocellular carcinoma. EMT, epithelial-to-mesenchymal transition. All values are mean ± SD. ** vs NC group, p < 0.01

Mentions: E-cadherin is a marker of epithelial cells, and MMP-2 and MMP-9 are markers of mesenchymal cells. Their changes correlated with EMT [14]. We examined the expression of EMT markers (Snail, Slug, N-cadherin, Vimentin, and E-cadherin), MMP-2, and MMP-9 in HCCLM3 and Huh7 cells. The results showed the expression of Snail, Slug, N-cadherin, Vimentin, MMP-2, and MMP-9 in HCCLM3 cells decreased after the knockdown of miR-181a, while the expression of E-cadherin increased. In Huh7 cells, we found increased expression of Snail, Slug, N-cadherin, Vimentin, MMP-2, and MMP-9 but E-cadherin expression decreased after transfection with miR-181a mimics (Fig. 4a). Many studies have shown that GSK3b is a target of PI3K/Akt and can regulate MMPs [15]. We examined the effect of miR-181a on AKT signaling pathway and found that knockdown of miR-181a significantly decreased the expression of phosphorylated AKT and mTOR in HCCLM3 cells, whereas overexpression of miR-181a significantly increased the expression of phosphorylated AKT and mTOR in Huh7 cells (Fig. 4b).Fig. 4


Long non-coding RNA XIST regulates PTEN expression by sponging miR-181a and promotes hepatocellular carcinoma progression
MiR-181a promoted HCC cell invasion and EMT by activating AKT signaling. a The protein expression of EMT related genes (Snail, Slug, N-cadherin, Vimentin and E-cadherin), MMP-2, and MMP-9. b Western blot analysis of phosphorylated (active) AKT and mTOR in HCC cells transfected with miR-181a mimics or inhibitor. HCC, hepatocellular carcinoma. EMT, epithelial-to-mesenchymal transition. All values are mean ± SD. ** vs NC group, p < 0.01
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5383949&req=5

Fig4: MiR-181a promoted HCC cell invasion and EMT by activating AKT signaling. a The protein expression of EMT related genes (Snail, Slug, N-cadherin, Vimentin and E-cadherin), MMP-2, and MMP-9. b Western blot analysis of phosphorylated (active) AKT and mTOR in HCC cells transfected with miR-181a mimics or inhibitor. HCC, hepatocellular carcinoma. EMT, epithelial-to-mesenchymal transition. All values are mean ± SD. ** vs NC group, p < 0.01
Mentions: E-cadherin is a marker of epithelial cells, and MMP-2 and MMP-9 are markers of mesenchymal cells. Their changes correlated with EMT [14]. We examined the expression of EMT markers (Snail, Slug, N-cadherin, Vimentin, and E-cadherin), MMP-2, and MMP-9 in HCCLM3 and Huh7 cells. The results showed the expression of Snail, Slug, N-cadherin, Vimentin, MMP-2, and MMP-9 in HCCLM3 cells decreased after the knockdown of miR-181a, while the expression of E-cadherin increased. In Huh7 cells, we found increased expression of Snail, Slug, N-cadherin, Vimentin, MMP-2, and MMP-9 but E-cadherin expression decreased after transfection with miR-181a mimics (Fig. 4a). Many studies have shown that GSK3b is a target of PI3K/Akt and can regulate MMPs [15]. We examined the effect of miR-181a on AKT signaling pathway and found that knockdown of miR-181a significantly decreased the expression of phosphorylated AKT and mTOR in HCCLM3 cells, whereas overexpression of miR-181a significantly increased the expression of phosphorylated AKT and mTOR in Huh7 cells (Fig. 4b).Fig. 4

View Article: PubMed Central - PubMed

ABSTRACT

Background: Tumor metastasis often occurs in hepatocellular carcinoma (HCC) and influences the patient&rsquo;s prognosis, and microRNAs are reported to play key roles in tumor metastasis. This study was conducted to explore the effect of microRNAs on HCC metastasis.

Methods: The levels of miR-181a in HCC tissues, adjacent tissues, metastatic HCC tissues, and non-metastatic HCC tissues at different stages were determined by qRT-PCR. Effect of miR-181a on the proliferation, invasion, and metastasis of HCC cells was estimated by cell counting kits-8 (CCK-8), wound-healing, and Transwell assays. Software analysis and luciferase assays were used to explore the target gene of miR-181a.

Results: MiR-181a was up-regulated in HCC tissues and its expression level in metastatic HCC tissues was much higher than in non-metastasis samples. PTEN was found to be a target gene of miR-181a. MiR-181a had multiple binding sites with the long non-coding RNA (lncRNA) XIST. The regulation of miR-181a on PTEN was mediated by lncRNA XIST. The proliferation and invasion of cells with siXIST were significantly enhanced compared with those of control cells, while knockdown of miR-181a abolished the enhancing effects.

Conclusions: MiR-181a can promote HCC metastasis by targeting PTEN, which is regulated by lncRNA XIST.

Electronic supplementary material: The online version of this article (doi:10.1186/s12885-017-3216-6) contains supplementary material, which is available to authorized users.

No MeSH data available.


Related in: MedlinePlus