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Urokinase-type plasminogen activator receptor regulates apoptotic sensitivity of colon cancer HCT116 cell line to TRAIL via JNK-p53 pathway

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Abstract

The urokinase-type plasminogen activator receptor (uPAR) serves not only as an anchor for urokinase-type plasminogen activator but also participates in intracellular signal transduction events. In this study, we investigated whether uPAR could modulate TRAIL-induced apoptosis in human colon cancer cells HCT116. Using an antisense strategy, we established a stable HCT116 cell line with down-regulated uPAR. The sensitivity to TRAIL-induced apoptosis was evaluated by FACS analysis. Our results show that the inhibition of uPAR could sensitize HCT116 to TRAIL-induced apoptosis. uPAR inhibition changed the expression of mitochondrial apoptotic pathway proteins, including Bcl-2, Bax, Bid and p53, in a pro-apoptotic manner. We also found that the inhibition of uPAR down-regulated the phosphorylation of FAK, ERK and JNK. The inhibition of p53 by RNA interference rescued cells from enhanced apoptosis, thus indicating that p53 is critical for enhancing TRAIL-induced apoptosis. Furthermore, JNK, but not ERK, inhibition involved in the up-regulation of p53. JNK negatively regulated p53 protein level. Overall, our results show that uPAR inhibition can sensitize colon cancer cells HCT116 to TRAIL-induced apoptosis via active p53 and mitochondrial apoptotic pathways that JNK inhibition is involved.

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Abbreviations

uPAR:

Urokinase-type plasminogen activator receptor

uPA:

Urokinase-type plasminogen activator

AS:

Antisense

TRAIL:

Tumor necrosis factor (TNF)-related apoptosis-inducing ligand

DR4:

Death receptor 4

DR5:

Death receptor 5

ECM:

Extracellular matrix

FAK:

Focal adhesion kinase

ERK:

Extracellular signal-regulated kinases

JNK:

Jun N-terminal kinase

Bcl-2:

B cell lymphoma 2

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Acknowledgments

The authors are grateful to grants from the Ministry of Science and Technology (2012CB967004, 2014CB744501, 2012AA020304 and 2012ZX09401012), the Chinese National Natural Sciences Foundation (81121062, 31200572, 31070706 and 31071196), the Jiangsu Provincial Nature Science Foundation (BE2013630 and BZ2012050), the Bureau of Science and Technology of Changzhou, Jiangsu, China (CZ20130011, CE20135013, CZ20120004, CM20122003 and WF201207).

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The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, 22 Han Kou Road, Nanjing, 210093, People’s Republic of China

    Xiufeng Liu, Fan Qiu, Zhipeng Liu, Yan Lan, Kai Wang, Yao Wang & Zi-Chun Hua

  2. Changzhou High-Tech Research Institute of Nanjing University and Jiangsu Target Pharma Laboratories Inc., Changzhou, 213164, People’s Republic of China

    Zi-Chun Hua

  3. Department of Radiation Toxicology and Oncology, Beijing Institute of Radiation Medicine, 27 Tai Ping Road, Hai Dian District, Beijing, 100850, People’s Republic of China

    Ping-Kun Zhou

  4. Division of Critical Care and Surgery, St. George Hospital, University of New South Wales, Sydney, NSW, 2217, Australia

    Yao Wang

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  1. Xiufeng Liu
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Correspondence to Yao Wang or Zi-Chun Hua.

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Liu, X., Qiu, F., Liu, Z. et al. Urokinase-type plasminogen activator receptor regulates apoptotic sensitivity of colon cancer HCT116 cell line to TRAIL via JNK-p53 pathway. Apoptosis 19, 1532–1544 (2014). https://doi.org/10.1007/s10495-014-1025-9

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