Tumor Biology

, Volume 36, Issue 5, pp 3995–4003 | Cite as

Silencing clusterin gene transcription on effects of multidrug resistance reversing of human hepatoma HepG2/ADM cells

  • Wenjie Zheng
  • Wenli Sai
  • Min Yao
  • Hongbin Gu
  • Yao Yao
  • Qi Qian
  • Dengfu Yao
Research Article

Abstract

Abnormal clusterin (CLU) expression is associated with multidrug resistance (MDR) of hepatocellular carcinoma (HCC). In the present study, the CLU expression was analyzed in human hepatoma cells and chemoresistant counterpart HepG2/ADM cells. Compared with L02 cells, the overexpression of cellular CLU was identified in HepG2, HepG2/ADM, SMMC7721, Hep3B ,and PLC cells and relatively lower expression in Bel-7404, SNU-739, and MHCC97H cells. Specific short hairpin RNAs (shRNAs) to silence CLU gene transcription were designed, and the most effective sequences were screened. After the HepG2/ADM cells transfected with shRNA-1, the inhibition of CLU expression was 73.68 % at messenger RNA (mRNA) level by real-time quantitative RT-PCR with obvious enhancement in cell chemosensitivity, increasing apoptosis induced by doxorubicin using fluorescence kit, and Rh-123 retention qualified with flow cytometry. Knockdown CLU also significantly decreased the drug efflux pump activity through the depression of MDR1/P-glycoprotein (q = 11.739, P < 0.001). Moreover, silencing CLU led to downregulation of β-catenin (q = 13.544, P = 0.001), suggesting that downregulation of CLU might be a key point to reverse multidrug resistance of HepG2/ADM cells.

Keywords

Hepatocellular carcinoma Clusterin Multidrug resistance Wnt/β-catenin 

Abbreviations

A

Absorbance

ABC

ATP-binding cassette

CLU

Clusterin

HCC

Hepatocellular carcinoma

MDR

Multidrug resistance

nCLU

Nuclear clusterin

PBS

Phosphate buffered saline

RT-PCR

Reverse transcriptase-PCR

sCLU

Secretory clusterin

shRNA

Short hairpin RNA

Wnt

Wnt/β-catenin signaling pathway

Notes

Acknowledgments

This work was supported by the Grants from the National Natural Science Foundation (81200634), the Projects of Jiangsu Medical Science (PADA, 2013-WSW-011, 2014-YY-028, HK201102, and BL2012053) and the Qinglan Project of Jiangsu Higher Education, and the International S&T Cooperation Program (2013DFA32150) of China

Conflicts of interest

None

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Wenjie Zheng
    • 1
  • Wenli Sai
    • 1
  • Min Yao
    • 2
  • Hongbin Gu
    • 3
  • Yao Yao
    • 3
  • Qi Qian
    • 4
  • Dengfu Yao
    • 1
  1. 1.Research Center of Clinical MedicineAffiliated Hospital of Nantong UniversityNantongChina
  2. 2.Medical School of Nantong UniversityNantongChina
  3. 3.The Hospital of Nantong Maternal and Childcare ServiceNantongChina
  4. 4.Center of OncologyAffiliated Hospital of Nantong UniversityNantongChina

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