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

Zu G, Wang H, Wang J, Dou Y, Zhao W, Sun Y - Exp Ther Med (2014)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250013, P.R. China.

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.

No MeSH data available.


Related in: MedlinePlus

Influence of RPTI on BGC-823 cell shape (magnification, ×100). (A) Control; (B) RPTI treatment (10 μg/ml RPTI, 72 h after addition to the cells). RPTI, Rhizoma Pinelliae trypsin inhibitor.
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f2-etm-08-01-0248: Influence of RPTI on BGC-823 cell shape (magnification, ×100). (A) Control; (B) RPTI treatment (10 μg/ml RPTI, 72 h after addition to the cells). RPTI, Rhizoma Pinelliae trypsin inhibitor.

Mentions: The BGC-823 cells were treated with 10 μg/ml RPTI (Fig. 2). After 72 h, the changes in cell shape were observed under an inverted microscope. The BGC-823 cells in the RPTI group were less shrunken and were significantly reduced in number compared with those of the control group, with numerous suspended cells being dead. The cells exhibited nuclear pyknosis and rough cytoplasms and underwent less fission. The control cells had regular shapes and good adherent growth.


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

Zu G, Wang H, Wang J, Dou Y, Zhao W, Sun Y - Exp Ther Med (2014)

Influence of RPTI on BGC-823 cell shape (magnification, ×100). (A) Control; (B) RPTI treatment (10 μg/ml RPTI, 72 h after addition to the cells). RPTI, Rhizoma Pinelliae trypsin inhibitor.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4061196&req=5

f2-etm-08-01-0248: Influence of RPTI on BGC-823 cell shape (magnification, ×100). (A) Control; (B) RPTI treatment (10 μg/ml RPTI, 72 h after addition to the cells). RPTI, Rhizoma Pinelliae trypsin inhibitor.
Mentions: The BGC-823 cells were treated with 10 μg/ml RPTI (Fig. 2). After 72 h, the changes in cell shape were observed under an inverted microscope. The BGC-823 cells in the RPTI group were less shrunken and were significantly reduced in number compared with those of the control group, with numerous suspended cells being dead. The cells exhibited nuclear pyknosis and rough cytoplasms and underwent less fission. The control cells had regular shapes and good adherent growth.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250013, P.R. China.

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.

No MeSH data available.


Related in: MedlinePlus