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. 2019 May 24:12:4099-4107.
doi: 10.2147/OTT.S185156. eCollection 2019.

Long non-coding RNA H19 promotes the proliferation and invasion of lung cancer cells and regulates the expression of E-cadherin, N-cadherin, and vimentin

Shu Liao  1 , Chaxiu Yu  2 , Hucheng Liu  3 , Congkai Zhang  1 , Yong Li  1 , Xiaojun Zhong  1
Affiliations

Long non-coding RNA H19 promotes the proliferation and invasion of lung cancer cells and regulates the expression of E-cadherin, N-cadherin, and vimentin

Shu Liao et al. Onco Targets Ther. .

Abstract

Background: This study aimed to explore the effect of long non-coding RNA (LncRNA) H19 on the proliferation and invasion of lung carcinoma cells A549, and to determine its molecular targets.

Methods: A549 cells were with either LncRNA H19 or LncRNA H19 shRNA, and the expression levels of LncRNA H19 were evaluated by quantitative real-time PCR (RT-PCR). We measured cell proliferation using the CCK-8 assay, cell counting assays, and colony formation assay in response to shLncRNA H19-2. Cell migration and invasion were assessed by wound healing assay and Transwell assay, respectively. The mRNA and protein expression levels of E-cadherin, N-cadherin, and vimentin were determined by RT-PCR and western blot, respectively.

Results: The three LncRNA H19 shRNAs used in our study significantly reduced the expression levels of LncRNA H19 in A549 cells (P<0.05). Moreover, LncRNA H19 shRNA 2 (shLncRNA-2) was the most potent inhibitor of LncRNA H19 expression, and was selected for further experimentation. Transfection with shLncRNA H19-2 significantly decreased the proliferation, migration, and invasion of A549 cells, while overexpression of LncRNA H19 had the opposite effect in these cells (P<0.05). In response to shLncRNA H19-2, the expression levels of E-cadherin were notably elevated (P<0.05), while the expression levels of N-cadherin and vimentin were decreased (P<0.05). In contrast, overexpression of LncRNA H19 induced the expression of E-cadherin, and blocked the expression of N-cadherin, and vimentin (P<0.05).

Conclusion: Our results suggest that LncRNA H19 mediates the proliferation and invasion of lung cancer cells via upregulation of N-cadherin and vimentin, and downregulation of E-cadherin.

Keywords: E-cadherin; N-cadherin; long non-coding RNA; lung cancer; vimentin.

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Conflict of interest statement

Disclosure The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
mRNA expression levels of LncRNA H19 in A549 cells after transfection with various LncRNA H19 silencing constructs, as measured by RT-PCR. Notes: Treatment with shLncRNA H19-2 resulted in the lowest mRNA expression levels of LncRNA H19 compared with all other constructs. *P<0.05 vs the control group. Abbreviations: RT-PCR, reverse transcription PCR; LncRNA, long non-coding RNA.
Figure 2
Figure 2
mRNA expression levels of LncRNA H19 in A549 cells transfected with different LncRNA H19 gene vectors, as measured by RT-PCR. Notes: Transfection with shLncRNA H19 significantly reduced the mRNA expression levels of LncH19 in A549 cells, while LncRNA H19 significantly elevated the mRNA expression levels compared with the control group. *P<0.05 vs the control group. Abbreviations: RT-PCR, reverse transcription PCR; LncRNA, long non-coding RNA.
Figure 3
Figure 3
The proliferation of A549 cells transfected with different LncRNA H19 gene vectors, as assessed by CCK-8 assay (A), cell counting (B), and colony formation (C). Notes: shLncRNA H19 decreased the proliferation of A549 cells, while LncRNA H19 increased their proliferation compared with the control group. *P<0.05 vs the control group. Abbreviations: LncRNA, long non-coding RNA; CCK-8, Cell Counting Kit-8.
Figure 4
Figure 4
(A) The migration of A549 cells transfected with different LncRNA H19 gene vectors, as evaluated by wound healing assay. (B) Qualitative distribution and quantitative number of A549 cells transfected with different LncRNA H19 gene vectors in a typical visual field of the lower chamber, as evaluated by transwell assay. Compared with the control group, the migration and invasion of A549 cells was significantly reduced by shLncRNA H19 and was increased by LncRNA H19. *P<0.05 vs the control group. Abbreviation: LncRNA, long non-coding RNA.
Figure 4
Figure 4
(A) The migration of A549 cells transfected with different LncRNA H19 gene vectors, as evaluated by wound healing assay. (B) Qualitative distribution and quantitative number of A549 cells transfected with different LncRNA H19 gene vectors in a typical visual field of the lower chamber, as evaluated by transwell assay. Compared with the control group, the migration and invasion of A549 cells was significantly reduced by shLncRNA H19 and was increased by LncRNA H19. *P<0.05 vs the control group. Abbreviation: LncRNA, long non-coding RNA.
Figure 5
Figure 5
mRNA and protein levels of E-cadherin, N-cadherin, and vimentin in A549 cells transfected with different LncRNA H19 gene vectors, as assessed by RT-PCR and Western blot, respectively. Notes: In response to shLncRNA H19, the expression levels of E-cadherin were significantly increased, and the expression levels of N-cadherin and vimentin were reduced. An opposite effect with regards to the expression levels of E-cadherin, N-cadherin, and vimentin was observed with LncRNA H19 overexpression. *P<0.05 vs the control group. Abbreviations: RT-PCR, reverse transcription PCR; LncRNA, long non-coding RNA.
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