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LncRNA PVT1 promotes proliferation and invasion through enhancing Smad3 expression by sponging miR-140-5p in cervical cancer

Qing-Qing Chang
Qing-Qing Chang
, 
Chun-Yan Chen
Chun-Yan Chen
, 
Zhao Chen
Zhao Chen
 oraz 
Shuai Chang
Shuai Chang
   | 18 paź 2019
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Article Category: Research Article
Data publikacji: 18 paź 2019
Zakres stron: 443 - 452
Otrzymano: 23 maj 2019
Przyjęty: 21 sie 2019
DOI: https://doi.org/10.2478/raon-2019-0048
© 2019 Qing-Qing Chang, Chun-Yan Chen, Zhao Chen, Shuai Chang, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

Expressions of PVT1, miR-140-5p and Smad3 in normal cervical epithelial cells and cervical cancer cell lines. Expression levels of PVT1 (A), miR-140-5p (B) and Smad3 (C) were detected by qRT-PCR. (D) Protein expressions of Smad3 were detected by western blotting. T he data are presented as means ± SD of three independent experiments. Statistical significance compared with the normal cervical epithelial cells is indicated by *P < 0.05 and **P < 0.01.
Expressions of PVT1, miR-140-5p and Smad3 in normal cervical epithelial cells and cervical cancer cell lines. Expression levels of PVT1 (A), miR-140-5p (B) and Smad3 (C) were detected by qRT-PCR. (D) Protein expressions of Smad3 were detected by western blotting. T he data are presented as means ± SD of three independent experiments. Statistical significance compared with the normal cervical epithelial cells is indicated by *P < 0.05 and **P < 0.01.

Figure 2

MiR-140-5p was a target of lncRNA PVT1 and Smad3 was a downstream target of miR-140-5p. (A) Binding sites between lncRNA PVT1 and miR-140-5p. (B) Binding sites between miR-140-5p and Smad3. Dual-luciferase assay was applied to explore the interaction between PVT1 and miR-140-5p in HeLa cells (C) and SiHa cells (D). Dual-luciferase assay was applied to explore the interaction between miR-140-5p and Smad3 in HeLa cells (E) and SiHa cells (F). The data are presented as means ± SD of three independent experiments. Statistical significance compared with miR-NC is indicated by *P < 0.05 and **P < 0.01.
MiR-140-5p was a target of lncRNA PVT1 and Smad3 was a downstream target of miR-140-5p. (A) Binding sites between lncRNA PVT1 and miR-140-5p. (B) Binding sites between miR-140-5p and Smad3. Dual-luciferase assay was applied to explore the interaction between PVT1 and miR-140-5p in HeLa cells (C) and SiHa cells (D). Dual-luciferase assay was applied to explore the interaction between miR-140-5p and Smad3 in HeLa cells (E) and SiHa cells (F). The data are presented as means ± SD of three independent experiments. Statistical significance compared with miR-NC is indicated by *P < 0.05 and **P < 0.01.

Figure 3

Silencing of lncRNA PVT1 inhibited proliferation of cervical cancer cells. Both HeLa and SiHa cells were transfected with sh-NC or sh-PVT1. Expressions of PVT1 (A, B) and miR-140-5p (C, D) were determined by qRT-PCR. (E-F) mRNA levels and (G, H) proteins levels of Smad3 were determined by qRT-PCR and western blotting respectively. (I, J) Colony formation assays were performed to evaluate the proliferation of both HeLa and SiHa cells. The data are presented as means ± SD of three independent experiments. Statistical significance compared with sh-NC is indicated by *P < 0.05 and **P < 0.01.
Silencing of lncRNA PVT1 inhibited proliferation of cervical cancer cells. Both HeLa and SiHa cells were transfected with sh-NC or sh-PVT1. Expressions of PVT1 (A, B) and miR-140-5p (C, D) were determined by qRT-PCR. (E-F) mRNA levels and (G, H) proteins levels of Smad3 were determined by qRT-PCR and western blotting respectively. (I, J) Colony formation assays were performed to evaluate the proliferation of both HeLa and SiHa cells. The data are presented as means ± SD of three independent experiments. Statistical significance compared with sh-NC is indicated by *P < 0.05 and **P < 0.01.

Figure 4

Silencing of lncRNA PVT1 suppressed the metastasis of cervical cancer cells. Both HeLa and SiHa cells were transfected with sh-NC or sh-PVT1. (A-D) Wound healing assay was conducted to evaluate the migration activities of HeLa and SiHa cells. (E-H) Transwell assay was conducted to evaluate the invasion activities of HeLa and SiHa cells. (I, J) Expressions of E-cadherin, N-cadherin, vimentin and Snail were determined by western blotting in both HeLa and SiHa cells. The data are presented as means ± SD of three independent experiments. Statistical significance compared with sh-NC is indicated by *P < 0.05 and **P < 0.01.
Silencing of lncRNA PVT1 suppressed the metastasis of cervical cancer cells. Both HeLa and SiHa cells were transfected with sh-NC or sh-PVT1. (A-D) Wound healing assay was conducted to evaluate the migration activities of HeLa and SiHa cells. (E-H) Transwell assay was conducted to evaluate the invasion activities of HeLa and SiHa cells. (I, J) Expressions of E-cadherin, N-cadherin, vimentin and Snail were determined by western blotting in both HeLa and SiHa cells. The data are presented as means ± SD of three independent experiments. Statistical significance compared with sh-NC is indicated by *P < 0.05 and **P < 0.01.

Figure 5

Silencing of miR-140-5p reversed the effects of PVT1 inhibition on the proliferation of cervical cancer cells. Both HeLa and SiHa cells were transfected with or without miR-140-5p inhibitor in the presence of sh-PVT1. (A, B) mRNA expressions and (C, D) protein expressions of Smad3 were determined by qRT-PCR and western blotting in both HeLa and SiHa cells. (E, F) Colony formation assays were performed to evaluate the proliferation of both HeLa and SiHa cells. The data are presented as means ± SD of three independent experiments. Statistical significance compared with sh-PVT1 is indicated by *P < 0.05 and **P < 0.01.
Silencing of miR-140-5p reversed the effects of PVT1 inhibition on the proliferation of cervical cancer cells. Both HeLa and SiHa cells were transfected with or without miR-140-5p inhibitor in the presence of sh-PVT1. (A, B) mRNA expressions and (C, D) protein expressions of Smad3 were determined by qRT-PCR and western blotting in both HeLa and SiHa cells. (E, F) Colony formation assays were performed to evaluate the proliferation of both HeLa and SiHa cells. The data are presented as means ± SD of three independent experiments. Statistical significance compared with sh-PVT1 is indicated by *P < 0.05 and **P < 0.01.

Figure 6

Silencing of miR-140-5p reversed the effects of PVT1 inhibition on the metastasis of cervical cancer cells. (A-D) Wound healing assay was conducted to evaluate the migration activities of HeLa and SiHa cells. (E-H) Transwell assay was conducted to evaluate the invasion activities of HeLa and SiHa cells. (I, J) Expressions of E-cadherin, N-cadherin, vimentin and Snail were determined by western blotting in both HeLa and SiHa cells. The data are presented as means ± SD of three independent experiments. Statistical significance compared with sh-PVT1 is indicated by *P < 0.05 and **P < 0.01.
Silencing of miR-140-5p reversed the effects of PVT1 inhibition on the metastasis of cervical cancer cells. (A-D) Wound healing assay was conducted to evaluate the migration activities of HeLa and SiHa cells. (E-H) Transwell assay was conducted to evaluate the invasion activities of HeLa and SiHa cells. (I, J) Expressions of E-cadherin, N-cadherin, vimentin and Snail were determined by western blotting in both HeLa and SiHa cells. The data are presented as means ± SD of three independent experiments. Statistical significance compared with sh-PVT1 is indicated by *P < 0.05 and **P < 0.01.
eISSN:
1581-3207
Język:
Angielski
Częstotliwość wydawania:
4 razy w roku
Dziedziny czasopisma:
Medicine, Clinical Medicine, Internal Medicine, Haematology, Oncology, Radiology
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