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. 2022 Apr;13(4):8302-8310.
doi: 10.1080/21655979.2022.2038449.

Long non-coding RNA NCK1-AS1 is overexpressed in esophageal squamous cell carcinoma and predicts survival

Xin Fu  1 , Xi Chen  1 , Yuanyuan Si  1 , Youjie Yao  1 , Zhengming Jiang  1 , Kui Chen  1
Affiliations

Long non-coding RNA NCK1-AS1 is overexpressed in esophageal squamous cell carcinoma and predicts survival

Xin Fu et al. Bioengineered. 2022 Apr.

Abstract

Long noncoding RNAs have shown pivotal regulatory roles in tumorigenesis and progression. NCK1-AS1 promotes cervical cancer, while its involvement in esophageal cancer is hardly known. This study enrolled 52 esophageal squamous cell carcinoma (ESCC) patients (30 males and 22 females) at the average age of 56.4 ± 6.6 years in the range from 46 to 70 years, explored the involvement of NCK1-AS1 in ESCC, and analyzed the possible interaction between NCK1-AS1 and TGF-β signaling. Changes in gene expression were analyzed using RT-qPCR and Western blot. Interactions between gene expressions were analyzed using ESCC cells with transient transfections. Cell invasion and migration were analyzed using Transwell assays. Our data showed that plasma NCK1-AS1 was overexpressed in ESCC patients and positively correlated with NCK1-AS1 expression in tumor tissues but not in non-tumor tissues. Moreover, high plasma NCK1-AS1 levels were accompanied with poor survival. TGF-β1 expression level was also increased in tumor tissues compared to the adjacent normal tissues and positively correlated with NCK1-AS1 in tumor tissues. TGF-β1 overexpression in ESCC cells did not affect NCK1-AS1 expression, while NCK1-AS1 overexpression in ESCC cells upregulated TGF-β1. Moreover, TGF-β1 and NCK1-AS1 overexpression increased ESCC cell migration and invasion, while TGF-β inhibitor reduced the effects of NCK1-AS1 overexpression. Overall, NCK1-AS1 may promote ESCC by upregulating TGF-β1.

Keywords: Esophageal squamous cell carcinoma; TGF-β1; lncRNANCK1-AS1; regulation; survival.

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Conflict of interest statement

No potential conflict of interest was reported by the author(s).

Figures

None
Graphical abstract
Figure 1.
Figure 1.
NCK1-AS1 was upregulated in ESCC tissues and positively correlated with its plasma level. Expression data analyzed by paired t-test showed that NCK1-AS1 expression was significantly upregulated in ESCC (a). * p < 0.05. Linear regression showed that plasma NCK1-AS1 levels were positively and significantly correlated with its levels in ESCC tissues (b) but not in adjacent non-cancer tissues (c).
Figure 2.
Figure 2.
High plasma NCK1-AS1 levels were correlated with poor survival. Overall survival condition of patients in the high NCK1-AS1 group was significantly worse than that of patients in the low NCK1-AS1 level group.
Figure 3.
Figure 3.
TGF-β1 mRNA expression was upregulated in ESCC tissues and positively correlated with NCK1-AS1. Luciferase reporter assay detected that TGF-β1 is related to NCK1-AS1 (a). Expression data analyzed by paired t-test showed that TGF-β1 mRNA expression was significantly upregulated in ESCC tissues compared to non-cancer tissues (b) (* p < 0.05). Linear regression showed that TGF-β1 and NCK1-AS1 were significantly and positively correlated in ESCC tissues (c) but not in adjacent non-cancer tissues (d).
Figure 4.
Figure 4.
NCK1-AS1 overexpression stimulated TGF-β1 expression. Our cells have undergone cell STR identification. Overexpression of TGF-β1 and NCK1-AS1 was confirmed at 24 h after transient transfections (a). TGF-β1 overexpression did not affect NCK1-AS1 (b), while NCK1-AS1 upregulated TGF-β1 expression in ESCC cells (c) (* p < 0.05).
Figure 5.
Figure 5.
NCK1-AS1 stimulated ESCC cell migration and invasion through TGF-β1. TGF-β1 and NCK1-AS1 overexpression increased ESCC cell migration (a) and invasion (b). In addition, TGF-β inhibitor SB431542 attenuated the effect of NCK1-AS1 overexpression. TGF-β1 and sh-NCK1-AS1 affected ESCC cell migration (c) and invasion (d) (* p < 0.05).
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