DOI QR코드

DOI QR Code

Silencing of Rac3 Inhibits Proliferation and Induces Apoptosis of Human Lung Cancer Cells

  • Liu, Tie-Qin (Department of Thoracic Surgery, First Affiliated Hospital, China Medical University) ;
  • Wang, Ge-Bang (Department of Thoracic Surgery, First Affiliated Hospital, China Medical University) ;
  • Li, Zheng-Jun (Department of Thoracic Surgery, Shenyang Chest Hospital) ;
  • Tong, Xiang-Dong (Department of Thoracic Surgery, General Hospital of Shenyang Military Area Command) ;
  • Liu, Hong-Xu (Department of Thoracic Surgery, First Affiliated Hospital, China Medical University)
  • Published : 2015.04.14

Abstract

Background: Rac3, a member of the Rac family of small guanosine triphosphatases (GTPases), regulates a variety of cell functions, including the organization of the cytoskeleton, cell migration, and invasion. Overexpression of Rac3 has been reported in several human cancers. However, the role of Rac3 in lung cancer (LC) has not been determined in detail. The purpose of this study was to investigate the effect of silencing of Rac3 expression in human LC cells and the consequences for cell survival. Materials and Methods: Lentivirus small hairpin RNA (shRNA) interference techniques were utilized to knock down the Rac3 gene. Gene and protein expression was quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. LC cell apoptosis was examined by annexin V-APC /propidium iodide staining. Results: Efficient silencing of Rac3 strongly inhibited A549 cell proliferation and colony formation ability, and significantly decreased tumor growth. Moreover, flow cytometry analysis showed that knockdown of Rac3 led to G2/M phase cell cycle arrest as well as an excess accumulation of cells in the G1 and S phase. Conclusions: Thus, functional analysis using shRNAs revealed a critical role for Rac3 in the tumor growth of LC cells. shRNA silencing of Rac3 could provide an effective strategy to treat LC.

Keywords

Lung cancer;Rac3;proliferation;apoptosis;shRNAs

Acknowledgement

Supported by : National Natural Science Foundation of China

References

  1. Allemani C, Weir HK, Carreira H, et al (2014). Global surveillance of cancer survival 1995-2009: analysis of individual data for 25676887 patients from 279 populationbased registries in 67 countries (CONCORD-2). Lancet. [Epub ahead of print].
  2. Baugher PJ, Krishnamoorthy L, Price JE, et al (2005). Rac1 and Rac3 isoform activation is involved in the invasive and metastatic phenotype of human breast cancer cells. Breast Cancer Res, 7, 965-74. https://doi.org/10.1186/bcr1329
  3. Chan AY, Coniglio SJ, Chuang YY, et al (2005). Roles of the Rac1 and Rac3 GTPases in human tumor cell invasion. Oncogene, 24, 7821-9. https://doi.org/10.1038/sj.onc.1208909
  4. Chatterjee M, Sequeira L, Jenkins-Kabaila M, et al (2011). Individual rac GTPases mediate aspects of prostate cancer cell and bone marrow endothelial cell interactions. J Signal Transduct, 541851.
  5. Chen QY, Xu LQ, Jiao DM, et al (2011). Silencing of Rac1 modifies LC cell migration, invasion and actin cytoskeleton rearrangements and enhances chemosensitivity to antitumor drugs. Int J Mol Med, 28, 769-76.
  6. Engers R, Ziegler S, Mueller M, et al (2007). Prognostic relevance of increased Rac GTPase expression in prostate carcinomas. Endocr Relat Cancer, 14, 245-56. https://doi.org/10.1677/ERC-06-0036
  7. Etienne-Manneville S, Hall A (2002). Rho GTPases in cell biology. Nature, 420, 629-35. https://doi.org/10.1038/nature01148
  8. Franken NA, Rodermond HM, Stap J, et al (2006). Clonogenic assay of cells in vitro. Nat Protoc, 1, 2315-9. https://doi.org/10.1038/nprot.2006.339
  9. Gest C, Joimel U, Huang L, et al (2013). Rac3 induces a molecular pathway triggering breast cancer cell aggressiveness: differences in MDA-MB-231 and MCF-7 breast cancer cell lines. BMC Cancer, 13, 63. https://doi.org/10.1186/1471-2407-13-63
  10. Haataja L, Groffen J, Heisterkamp N (1997). Characterization of Rac3, a novel member of the Rho family. J Biol Chem, 272, 20384-88. https://doi.org/10.1074/jbc.272.33.20384
  11. Lois C, Hong EJ, Pease S, et al (2002). Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science, 295, 868-72. https://doi.org/10.1126/science.1067081
  12. Mira JP, Benard V, Groffen J, et al (2000). Endogenous, hyperactive RAC3 controls proliferation of breast cancer cells by a p21-activated kinase-dependent pathway. Proc Natl Acad Sci USA, 97, 185-9.
  13. Niggli V, Schlicht D, Affentranger S (2009). Specific roles of Rac1 and Rac2 in motile functions of HT1080 fibrosarcoma cells. Biochem Biophys Res Commun, 386, 688-92. https://doi.org/10.1016/j.bbrc.2009.06.098
  14. Onesto C, Shutes A, Picard V, et al (2008). Characterization of EHT 1864, a novel small molecule inhibitor of Rac family small GTPases. Methods Enzymol, 439, 111-29. https://doi.org/10.1016/S0076-6879(07)00409-0
  15. Siegel R, Ma J, Zou Z, et al (2014). Cancer statistics, 2014. CA Cancer J Clin, 64, 9-29. https://doi.org/10.3322/caac.21208
  16. Walker MP, Zhang M, Le TP, et al (2011). RAC3 is a promigratory co-activator of ERalpha. Oncogene, 30, 1984-94. https://doi.org/10.1038/onc.2010.583
  17. Yuan K, Qian C, Zheng R (2009). Prognostic significance of immunohistochemical Rac1 expression in survival in early operable non-small cell LC. Med Sci Monit, 15, 313-9.

Cited by

  1. Rac3 regulates cell proliferation through cell cycle pathway and predicts prognosis in lung adenocarcinoma vol.37, pp.9, 2016, https://doi.org/10.1007/s13277-016-5126-7
  2. Novel recombinant protein FlaA N/C increases tumor radiosensitivity via NF-κB signaling in murine breast cancer cells vol.12, pp.4, 2016, https://doi.org/10.3892/ol.2016.4957
  3. Sex-Specific Associations between Particulate Matter Exposure and Gene Expression in Independent Discovery and Validation Cohorts of Middle-Aged Men and Women vol.125, pp.4, 2017, https://doi.org/10.1289/EHP370
  4. Prognostic Values of EPDR1 Hypermethylation and Its Inhibitory Function on Tumor Invasion in Colorectal Cancer vol.10, pp.10, 2018, https://doi.org/10.3390/cancers10100393