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Down-regulation of JAK1 by RNA interference inhibits growth of the lung cancer cell line A549 and interferes with the PI3K/mTOR pathway

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Abstract

Purpose

The mammalian Janus kinase (JAK) family plays a critical role in cytokine/growth factor signalling pathways and is associated with human cancers. In this study, we explored the role of JAK1 in the non-small cell lung cancer (NSCLC) cell line A549 and its molecular crosstalk with the phosphatidyl inositol-3-kinase (PI3K)/mammalian target of the rapamycin (mTOR) pathway.

Methods

One hundred and two NSCLC and 50 normal lung specimens were collected after surgical resection. JAK1 expression and phosphorylation were determined via immunohistochemical staining (IHC) assay. A stable knockdown of JAK1 was performed in A549 cells by RNA interference. Stable cell proliferation, cell cycle, apoptosis, and invasion were characterised in vitro. Tumourigenicity was analysed in vivo. The NSCLC xenograft protein expression of PI3K/mTOR pathway molecules was determined by Western blot assay.

Results

JAK1 expression was higher in NSCLC tissues than in normal lung tissues (P < 0.01). JAK1 knockdown in A549 cells significantly inhibited cell proliferation and invasion while promoting cell arrest at G0/G1 phase (all P < 0.05). The xenograft model showed that JAK1 suppression inhibited tumour growth compared with normal control (P < 0.05). Moreover, JAK1 knockdown inhibited mTOR or P70 ribosomal protein S6 kinase (P70S6K) phosphorylation, but increased glycogen synthase kinase-3α (GSK-3α) and B-cell lymphoma-extra large (Bcl-xl) phosphorylation. Total protein expression and Akt1/2 phosphorylated status remained unchanged.

Conclusion

Our study suggests that JAK1 expression and phosphorylation is abnormal in NSCLC tissues. The knockdown of JAK1 significantly inhibits tumourigenicity of the A549 cell line and demonstrates that crosstalk between the JAK1 and PI3K/mTOR pathways is involved.

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Acknowledgments

We thank the patients and their families for participating in our study for their patience and cooperation. This work was supported by grants from the National Basic Research Program of China (2009CB941200) and the Nature Science Foundation of China (grant 30771228 to H.X. and grant 30771227 to X.M.).

Conflict of interest

We declare that we have no conflicts of interest.

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Authors and Affiliations

  1. Department of Respiratory Medicine, West China Hospital, Sichuan University, Chengdu, 610041, People’s Republic of China

    Dan Liu, Jing Zeng, Bojiang Chen, Na Huang, Na Guo & Weimin Li

  2. Thoracic and Cardiovascular Surgery, West China Hospital, Sichuan University, Chengdu, China

    Lunxu Liu

  3. Laboratory of Stem Cell Biology, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, China

    Hong Xu & Xianming Mo

  4. Clinical Laboratory Department, Sichuan Academy of Medical Sciences and Sichuan Provincial People’s Hospital, Chengdu, China

    Yi Huang

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  1. Dan Liu
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  8. Hong Xu
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  10. Weimin Li
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Correspondence to Xianming Mo or Weimin Li.

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D. Liu and Y. Huang contributed equally to this work.

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Liu, D., Huang, Y., Zeng, J. et al. Down-regulation of JAK1 by RNA interference inhibits growth of the lung cancer cell line A549 and interferes with the PI3K/mTOR pathway. J Cancer Res Clin Oncol 137, 1629–1640 (2011). https://doi.org/10.1007/s00432-011-1037-6

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