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

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

Radiology and Oncology
Volume 53 (2019): Issue 4 (December 2019)
By: Qing-Qing Chang,  Chun-Yan Chen,  Zhao Chen and  Shuai Chang  
Open Access
|Oct 2019

Figures & Tables

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.
DOI: https://doi.org/10.2478/raon-2019-0048 | Journal eISSN: 1581-3207 | Journal ISSN: 1318-2099
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Language: English
Page range: 443 - 452
Submitted on: May 23, 2019
|
Accepted on: Aug 21, 2019
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Published on: Oct 18, 2019
Published by: Association of Radiology and Oncology
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
cervical cancer,
lncRNA PVT1,
miR-140-5p,
Smad3
Related subjects:
Medicine,
Clinical medicine,
Internal medicine,
Haematology, oncology,
Radiology

© 2019 Qing-Qing Chang, Chun-Yan Chen, Zhao Chen, Shuai Chang, published by Association of Radiology and Oncology
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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