Rhizoma Pinelliae trypsin inhibitor separation, purification and inhibitory activity on the proliferation of BGC‑823 gastric adenocarcinoma cells

Zu,Guohong; Wang,Houwei; Wang,Jie; Dou,Yan; Zhao,Weichong; Sun,Yuping

doi:10.3892/etm.2014.1701

Rhizoma Pinelliae trypsin inhibitor separation, purification and inhibitory activity on the proliferation of BGC‑823 gastric adenocarcinoma cells

Authors:
- Guohong Zu
- Houwei Wang
- Jie Wang
- Yan Dou
- Weichong Zhao
- Yuping Sun
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Affiliations: Department of Radiation Oncology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250013, P.R. China, Department of Chinese Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, Shandong 250013, P.R. China, Department of Surgery, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250013, P.R. China
Published online on: May 8, 2014 https://doi.org/10.3892/etm.2014.1701
Pages: 248-254

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Abstract

The aim of this study was to isolate and purify Rhizoma Pinelliae trypsin inhibitor (RPTI), determine its N‑terminal amino acid sequence and evaluate its inhibitory effect on the proliferation of poorly differentiated BGC‑823 human gastric adenocarcinoma cells. RPTI was separated and purified from a 40% (NH4)2SO4 precipitate of crude protein extract of Pinellia ternata tuber using affinity chromatography with trypsin as the ligand. The N‑terminal amino acid sequence of RPTI was determined using the Edman degradation method. The inhibitory effect of RPTI on BGC‑823 cell proliferation was detected in vitro using the MTT method and in vivo in tumour‑bearing mice. The purified RPTI showed a single band under SDS‑PAGE, its molecular weight was 14 kDa and its N‑terminal amino acid sequence was DPVVDG. RPTI inhibited trypsin activity, with an inhibition ratio of 1:6.78 (mass). RPTI significantly inhibited the proliferation of BGC‑823 cells in vitro. The IC50 of RPTI was 16.96 µg/ml within 48 h after treatment and 9.61 µg/ml within 72 h after treatment. Subcutaneous injection of RPTI around the tumour significantly inhibited BGC‑823 tumour growth in mice. The tumour inhibitory effect was concentration‑ and dose‑dependent. RPTI did not significantly influence the spleen coefficient of the mice. In conclusion, RPTI is a serine proteinase inhibitor with antitumour activity.

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