A potential human hepatocellular carcinoma inhibitor from Bauhinia purpurea L. seeds: from purification to mechanism exploration

Abstract

A 20-kDa Kunitz-type trypsin-chymotrypsin inhibitor, Bauhinia purpurea trypsin inhibitor (BPLTI), has been isolated from the seeds of B. purpurea L. by using liquid chromatography procedures that involved ion exchange chromatography on Sp-Sepharose and Mono S and gel filtration on Superdex 75. BPLTI demonstrated protease inhibitory activities of 7226 BAEE units/mg and 65 BTEE units/mg toward trypsin and α-chymotrypsin, respectively. BPLTI was relatively thermal (0–60°C) and pH (3–10) stable and its activity could be decreased by dithiothreitol treatment. BPLTI exhibited a wide spectrum of anti-proliferative and pro-apoptotic activities especially on human hepatocellular carcinoma Hep G2 cells. However, it was devoid of a significant antiproliferative effect on immortal human hepatic WRL 68 cells. We show here that BPLTI stimulates apoptosis in Hep G2 cells, including (1) evoking DNA damage including the production of chromatin condensation and apoptotic bodies; (2) induction of cell apoptosis/necrosis; (3) mitochondrial membrane depolarization; and (4) increasing the production of cytokines. Taken together, our findings show for the first time that purified protease inhibitor from B. purpurea L. seeds is a promising candidate for the treatment of human hepatocellular carcinoma.

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Abbreviations

BPLTI:

Bauhinia purpurea L. trypsin inhibitor

DTT:

Dithiothreitol

BAEE:

N-α-benzoyl-l-arginine ethyl ester

BTEE:

N-benzoyl-l-tyrosine ethyl ester

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Acknowledgments

We thank Dr. Carol P. Y. Lau, Prof. Lawrence Ramsden, and Prof. Richard M. K. Saunders from School of Biological Sciences, The University of Hong Kong, for their kind support of this project.

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The authors declare no conflict of interest.

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Correspondence to Xiu Juan Ye or Tzi Bun Ng.

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Fang, E.F., Bah, C.S.F., Wong, J.H. et al. A potential human hepatocellular carcinoma inhibitor from Bauhinia purpurea L. seeds: from purification to mechanism exploration. Arch Toxicol 86, 293–304 (2012). https://doi.org/10.1007/s00204-011-0751-9

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Keywords

  • Bauhinia purpurea L.
  • Trypsin inhibitor
  • Liver cancer
  • Cytokine
  • Apoptosis