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Fig. 7 | Cancer Cell International

Fig. 7

From: The lncRNA NEAT1 promotes the epithelial-mesenchymal transition and metastasis of osteosarcoma cells by sponging miR-483 to upregulate STAT3 expression

Fig. 7

The lncRNA NEAT1/miR-483/STAT3 axis contributes to the liver and lung metastases of osteosarcoma in mice. U2OS cells were transfected with the sh-NC or sh-NEAT1 plasmid. Subsequently, cells were subcutaneously injected into nude mice to establish liver and lung metastases of osteosarcoma (n = 5). Untreated nude mice served as the negative control. a Representative images of resected tumours from tumour-bearing mice. b The weight of resected tumours from tumour-bearing mice. c The tumour volume of tumours was monitored every 7 days for 6 weeks. d Representative images showing the distribution, quantity, and volume of metastatic osteosarcoma tumours in the lung (left panel) and liver (right panel) from different groups of mice, normal lung and liver tissues in mice without any treatment were served as the control. e H&E staining showing the invasion of metastatic osteosarcoma tumours, normal lung and liver tissues in mice without any treatment were served as the control. Scale bar: 50 μm. f The relative expression of NEAT1, miR-483 and STAT3 in primary tumour tissues and metastatic tumour tissues was examined using qRT-PCR. The NEAT1 and STAT3 mRNAs were normalized to the GAPDH mRNA, miR-483 was normalized to U6. Normal lung and liver tissues in mice without any treatment were served as the control in liver and lung panel. g The levels of STAT3 and STAT1 and the phosphorylation of STAT3 in primary tumour tissues and metastatic tumour tissues were assessed using western blotting. GAPDH was employed as the input control. Normal lung and liver tissues in mice without any treatment were served as the control in liver and lung panel. h The levels of N-cadherin, E-cadherin, Vimentin, and Snail in primary tumour tissues were assessed using western blotting. GAPDH was employed as the input control. i The levels of N-cadherin, E-cadherin, Vimentin, and Snail in metastatic tumour tissues in the lung and liver were assessed using western blotting. GAPDH was employed as the input control. All experiments described in this study were performed at least three times, which referred to biological replicates. Each biological replicate consisted of 3 wells with cells of the same passage, which referred to technical replicates. Error bars denoted mean ± SD. p values were calculated using unpaired two-tailed Student’s t-test or one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. ***p < 0.001, **p < 0.01, and *p < 0.05

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