Skip to main content

Advertisement

SpringerLink
Log in

Effect of short hairpin RNA of Bcl-xL gene on biological behaviors of HepG-2 cells

  • Original Paper
  • Published:
Medical Oncology Aims and scope Submit manuscript

Abstract

To investigate the inhibitory effect of short hairpin RNA (shRNA) targeting the Bcl-xL gene on the apoptosis and proliferation of hepatocellular carcinoma HepG-2 cells. We constructed Bcl-xL shRNA, and transfected it into HepG-2 cells, then detected Bcl-xL gene expression on mRNA and protein levels by reverse transcription–polymerase chain reaction (RT–PCR) and flow cytometry, respectively. Cell proliferation was detected by MTT assay. Cell apoptosis and Bcl-2 and Bax expression were detected by flow cytometry. The expression of Bcl-xL was obviously inhibited by RNA interference. The inhibition rates of Bcl-xL mRNA and protein were 86.6 and 70.2%, respectively. Bcl-xL shRNA inhibited cell proliferation (P < 0.05), whereas promoted apoptosis (P < 0.01). After transfection of Bcl-xL shRNA in HepG-2 cells, the expression of Bcl-2 did not change, but Bax increased significantly. Bcl-xL shRNA effectively inhibits Bcl-xL gene expression and proliferation of HepG-2 cells and promotes apoptosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Wong R, Frenette C. Updates in the management of hepatocellular carcinoma. Gastroenterol Hepatol (N Y). 2011;7(1):16–24.

    Google Scholar 

  2. Pazo Cid RA, Lao J, Lanzuela M. Doxorubicin plus sorafenib in treatment of advanced hepatocellular carcinoma. JAMA. 2011;305(8):781.

    Article  PubMed  Google Scholar 

  3. Scherr M, Morgan MA, Eder M. Gene silencing mediated by small interfering RNAs in mammalian cells. Curr Med Chem. 2003;10(3):245–56.

    PubMed  CAS  Google Scholar 

  4. Sawey ET, Chanrion M, Cai C, Wu G, Zhang J, Zender L, et al. Identification of a therapeutic strategy targeting amplified FGF19 in liver cancer by oncogenomic screening. Cancer Cell. 2011;19(3):347–58.

    Article  PubMed  CAS  Google Scholar 

  5. Broemer M, Tenev T, Rigbolt KT, Hempel S, Blagoev B, Silke J, et al. Systematic in vivo RNAi analysis identifies IAPs as NEDD8–E3 ligases. Mol Cell. 2010;40(5):810–22.

    Article  PubMed  CAS  Google Scholar 

  6. Danial NN. BCL-2 family proteins: critical checkpoints of apoptotic cell death. Clin Cancer Res. 2007;13(24):7254–63.

    Article  PubMed  CAS  Google Scholar 

  7. Suzuki H, Lee K, Matsuoka M. (2011) TDP-43-induced death is associated with altered regulation of BIM and BCL-XL and attenuated by caspase-mediated TDP-43 cleavage. J Biol Chem. [Epub ahead of print].

  8. Zhao WL, Daneshpouy ME. Prognostic significance of bcl-xL gene expression and apoptotic cell counts in follicular lymphoma. Blood. 2004;103(2):695–7.

    Article  PubMed  CAS  Google Scholar 

  9. Watanabe J, Kushihata F. Prognostic significance of Bcl-xL in human hepatocellular carcinoma. Surgery. 2004;135(6):604–12.

    Article  PubMed  Google Scholar 

  10. Watanabe J, Kushihata F, Honda K, Sugita A, Tateishi N, Mominoki K, et al. Prognostic significance of Bcl-xL in human hepatocellular carcinoma. Surgery. 2004;135(6):604–12.

    Article  PubMed  Google Scholar 

  11. Takehara T, Liu X, Fujimoto J, Friedman SL, Takahashi H. Expression and role of Bcl-xL in human hepatocellular carcinomas. Hepatology. 2001;34(1):55–61.

    Article  PubMed  CAS  Google Scholar 

  12. Xie YE, Tang EJ, Zhang DR, Ren BX. Down-regulation of Bcl-XL by RNA interference suppresses cell growth and induces apoptosis in human esophageal cancer cells. World J Gastroenterol. 2006;12(46):7472–7.

    PubMed  CAS  Google Scholar 

  13. Watanabe J, Kushihata F, Honda K, Mominoki K, Matsuda S, Kobayashi N. Bcl-xL overexpression in human hepatocellular carcinoma. Int J Oncol. 2002;21(3):515–9.

    PubMed  CAS  Google Scholar 

  14. Luo D, Cheng SC, Xie H, Xie Y. Effects of Bcl-2 and Bcl-XL protein levels on chemoresistance of hepatoblastoma HepG2 cell line. Biochem Cell Biol. 2000;78:119–26.

    PubMed  CAS  Google Scholar 

  15. Youle RJ, Strasser A. The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol. 2008;9(1):47–59.

    Article  PubMed  CAS  Google Scholar 

  16. Knudson CM, Brown NM. Mitochondria potential, bax “activation”, and programmed cell death. Methods Mol Biol. 2008;414:95–108.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The study was funded by scientific research project from education bureau of Liaoning province (No. 2008161).

Author information

Authors and Affiliations

  1. Department of Clinical Laboratory, The First Affiliated Hospital of Dalian Medical University, Dalian, 116011, China

    Duan Mengxi, Yuan Hong, Wang Nan, Li Shijun & Jiang Yanmei

Authors
  1. Duan Mengxi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuan Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wang Nan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Li Shijun
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jiang Yanmei
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yuan Hong.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mengxi, D., Hong, Y., Nan, W. et al. Effect of short hairpin RNA of Bcl-xL gene on biological behaviors of HepG-2 cells. Med Oncol 29, 1000–1004 (2012). https://doi.org/10.1007/s12032-011-9949-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12032-011-9949-1

Keywords

Search

Navigation