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LOX-1 is a poor prognostic indicator and induces epithelial-mesenchymal transition and metastasis in pancreatic cancer patients

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Abstract

Purpose

Pancreatic cancer (PC) is an aggressive type of cancer that exhibits a rapid progression. Previously LOX-1, which is a type II trans-membrane glycoprotein that is expressed in endothelial cells, has been found to be involved in the development of several types of cancer. As yet, however, the expression of LOX-1 and its functional consequences in PC have not been documented. The present study was aimed at investigating the prognostic relevance of LOX-1 expression in PC patients and at resolving its role in PC metastasis.

Methods

LOX-1 expression was assessed by immunohistochemistry on a tissue microarray containing samples from 98 PC patients. Kaplan-Meier analyses were performed to compare survival curves, whereas Cox regression analyses were performed to explore the independent prognostic value of LOX-1 expression on the overall survival (OS) of PC patients. Harrel’s concordance index was applied to calculate the predictive accuracy of established models. In addition, in vitro scratch wound healing and Transwell assays were used to assess the effect of LOX-1 expression silencing and over-expression on PC cell migration and invasion, whereas Cell Counting Kit-8 (CCK8) and Flow Cytometry (FCM) assays were used to assess its effects on PC cell proliferation and apoptosis.

Results

We found that LOX-1 is highly expressed in the PC tumor tissues tested and is related to the occurrence of lymph node metastases, higher TNM stages and a poor OS. We also found that LOX-1 expression may serve as an independent prognostic factor for the OS of PC patients. Our in vitro assays revealed that LOX-1 expression may promote the migration and invasion of PC cells through epithelial-mesenchymal transition (EMT). No effect on PC cell proliferation was noted.

Conclusions

From our data we conclude that a high LOX-1 expression in PC tissues is indicative for the occurrence of lymph node metastases, high TNM stages and a poor prognosis. LOX-1 may serve as an independent prognostic biomarker. Our in vitro assays additionally revealed that LOX-1 may enhance the migration and invasion of PC cells through EMT. LOX-1 may also serve as a novel therapeutic target.

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Acknowledgments

This work was supported by grants from the National Basic Research Program of China 973 Program (2012CB822104) and the National Natural Science Fund (31370808, 81572317, 31600648).

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Author notes

  1. Jie Zhang and Lei Zhang contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Glycoconjugate Research Ministry of Public Health, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, People’s Republic of China

    Jie Zhang, Can Li, Caiting Yang, Lili Li, Shushu Song, Hao Wu, Lan Wang & Jianxin Gu

  2. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, 138 Yixueyuan Road, Shanghai, 200032, People’s Republic of China

    Jie Zhang, Can Li, Caiting Yang, Lili Li, Shushu Song, Hao Wu, Lan Wang & Jianxin Gu

  3. Institute of Biomedical Sciences, Fudan University, Shanghai, 200032, People’s Republic of China

    Jie Zhang & Jianxin Gu

  4. Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai, 20032, People’s Republic of China

    Lei Zhang & Fenglin Liu

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  1. Jie Zhang
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Contributions

Jie Zhang designed and carried out experiments, analyzed data and wrote the manuscript; Lei Zhang collected and analyzed clinical data; Can Li, Caiting Yang and Lili Li performed experiments; Shushu Song and Hao Wu analyzed data; Jianxin Gu contributed the materials and experimental equipment; Lan Wang and Fenglin Liu conceived the research idea and took responsibility for this part of the project.

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Correspondence to Fenglin Liu or Lan Wang.

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Electronic supplementary material

Supplementary Fig. S1

ROC analysis for CES score of LOX-1 IHC staining. a ROC curve analysis shows the optimal cut-off value of CES is 6 (CES6), and the area under the ROC curve is 0.825 (95% CI, 0.733–0.916, P < 0.001). CES > 6 indicates high expression of LOX-1, while CES ≤ 6 indicates LOX-1 low expression. b Representative images show high and low expression of CLEC2, respectively. Scale bar, 100 mm. (GIF 660 kb)

High resolution (TIFF 8487 kb)

Supplementary Fig. S2

Survival analyses for PC patients in TNM I and TNM II-IV according to LOX-1 expression. Kaplan–Meier analyses of overall survival were performed in PC patients with early-stage cancer (TNM I) and advanced-stage cancer (TNM II-IV). (GIF 45 kb)

High resolution (TIFF 152 kb)

Supplementary Fig. S3

LOX-1 promotes migration of PC cells. Representative photographs of scratch wound-healing motility assays in AsPC-1 with LOX-1 overexpression or SW1990 cells with LOX-1 knock-down. (GIF 175 kb)

High resolution (TIFF 2817 kb)

Supplementary Fig. S4

LOX-1 has a positive correlation with IL-6 cytokine. a Correlation analysis of levels of different cytokines in LOX-1 knock-down cells and control cells by Real-time PCR. (GIF 69 kb)

High resolution (TIFF 355 kb)

Supplementary Table 1

(DOCX 12 kb)

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Zhang, J., Zhang, L., Li, C. et al. LOX-1 is a poor prognostic indicator and induces epithelial-mesenchymal transition and metastasis in pancreatic cancer patients. Cell Oncol. 41, 73–84 (2018). https://doi.org/10.1007/s13402-017-0360-6

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