Tumor Biology

, Volume 37, Issue 8, pp 10731–10743 | Cite as

The SMAD2/3 pathway is involved in hepaCAM-induced apoptosis by inhibiting the nuclear translocation of SMAD2/3 in bladder cancer cells

  • Xiaorong Wang
  • E. Chen
  • Min Tang
  • Xue Yang
  • Yin Wang
  • Zhan Quan
  • Xiaohou Wu
  • Chunli Luo
Original Article


The aim of this study was to explore the correlation between hepatocyte cell adhesion molecule (hepaCAM) and SMAD family member 2/3 (SMAD2/3) in bladder carcinoma, and the involvement of the SMAD2/3 pathway in hepaCAM-induced tumor apoptosis. Immunohistochemistry was used to measure hepaCAM and p-SMAD2/3 protein levels in bladder cancer tissues. Flow cytometry and Hoechst staining were used to study the effect of hepaCAM on cellular apoptosis. Western blot was employed to determine the expression of hepaCAM and SMAD2/3/caspase pathway molecules using a hepaCAM overexpression adenovirus, a caspase inhibitor (Z-VAD-FMK), and a SMAD2/3 activator (transforming growth factor (TGF)-β1), respectively. Translocation of p-SMAD2/3 was measured by immunofluorescence and western blot. HepaCAM proteins were significantly decreased (P < 0.05), while p-SMAD2/3 proteins were remarkably increased (P < 0.05) in bladder carcinoma compared to adjacent tissues. However, the low hepaCAM and high p-SMAD2/3 were not statistically associated with clinicopathological characteristics of the patients. A negative linear correlation between hepaCAM and p-SMAD2/3 was observed according to Pearson analysis (r = −0.712/−0.724, P = 0.008/0.011). Overexpression of hepaCAM activated caspase 3/8/9 and downregulated poly-ADP ribose polymerase (PARP) and p-SMAD2/3. Treatment of bladder cancer cells with Z-VAD-FMK + hepaCAM significantly downregulated procaspase 3/8/9 and PARP and induced cellular apoptosis, compared with that using Z-VAD-FMK alone. Similarly, combined treatment of TGF-β1 + hepaCAM significantly downregulated p-SMAD2/3, procaspase 3/8/9, and PARP and induced apoptosis of bladder cancer cells, compared with TGF-β1 alone. Overexpression of hepaCAM prevented the p-SMAD2/3 translocation from the cytoplasm to the nucleus in bladder cancer cells BIU-87 and T24. Our findings uncover that the p-SMAD2/3 pathway is critical for hepaCAM-induced cancer cell apoptosis and provide valuable insights for current and future Ad-hepaCAM and p-SMAD2/3 clinical trials.


Bladder cancer HepaCAM p-SMAD2/3 Caspase protein Cell apoptosis 



This work was conducted at the Department of Laboratory Diagnosis, Chongqing Medical University, and is supported by the National Natural Science Foundation of the People’s Republic of China (Grant No. 81072086).

Compliance with ethical standards

Written informed consent was received from all of the participants. This study was approved by the Ethics Committee of Chongqing Medical University.

Conflicts of interest



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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Xiaorong Wang
    • 1
  • E. Chen
    • 1
  • Min Tang
    • 1
  • Xue Yang
    • 1
  • Yin Wang
    • 1
  • Zhan Quan
    • 2
  • Xiaohou Wu
    • 2
  • Chunli Luo
    • 1
  1. 1.The Key Laboratory of Diagnostics Medicine, Ministry of EducationChongqing Medical UniversityChongqingChina
  2. 2.Department of UrologyThe First Affiliated Hospital of Chongqing Medical UniversityChongqingChina

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