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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

Department of Obstetrics and Gynecology, Hunan Provincial People’s HospitalChangsha, China
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Chang, Qing-Qing
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Chun-Yan Chen

Chun-Yan Chen

Department of Obstetrics and Gynecology, Hunan Provincial People’s HospitalChangsha, China
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Chen, Chun-Yan
, 
Zhao Chen

Zhao Chen

Department of Hematology, Hunan Provincial People’s HospitalChangsha, China
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Chen, Zhao
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Shuai Chang

Shuai Chang

Changsha Medical UniversityChangsha, China
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Chang, Shuai
  
Oct 18, 2019
LncRNA PVT1 promotes proliferation and invasion through enhancing Smad3 expression by sponging miR-140-5p in cervical cancer's Cover Image
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Article Category: Research Article
Published Online: Oct 18, 2019
Page range: 443 - 452
Received: May 23, 2019
Accepted: Aug 21, 2019
DOI: https://doi.org/10.2478/raon-2019-0048
Keywords
© 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.
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