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. 2002 Oct;8(5):832-6.
doi: 10.3748/wjg.v8.i5.832.

Apoptosis of hepatoma cells SMMC-7721 induced by Ginkgo biloba seed polysaccharide

Qun Chen  1 , Gui-Wen Yang, Li-Guo An
Affiliations

Apoptosis of hepatoma cells SMMC-7721 induced by Ginkgo biloba seed polysaccharide

Qun Chen et al. World J Gastroenterol. 2002 Oct.

Abstract

Aim: To study the apoptosis of hepatoma cells SMMC-7721 induced by polysaccharide isolated from Ginkgo biloba seed.

Methods: Ginkgo biloba seed polysaccharide (GBSP) was isolated by ethanol fractionation of Ginkgo biloba seed and purified by Sephadex G-200 chromatography. The purity of GBSP was verified by reaction with iodine-potassium iodide and ninhydrin and confirmed by UV spectrophotometer, cellulose acetate membrane electrophoresis and Sepharose 4B gel filtration chromatography. The Scanning Electron Microscope (SEM) and Flow Cytometry (FCM) were used to examine the SMMC-7721 cells with and without GBSP treatment at 500 mg/ml for 36 h.

Results: GBSP product obtained was of high purity with the average molecular weight of 1.86 X 10(5). Quantitative analysis of SMMC-7721 cells in vitro with FCM showed that the percentages of G(2)-M cells without and with GBSP treatment were 17.01+/-1.28 % and 11.77+/-1.50% (P<0.05), the debris ratio of the cells were 0.46+/-0.12 % and 0.06+/-0.06 % (P<0.01), and the apoptosis ratio of cells was 3.84+/-0.55 % and 9.13+/-1.48 % (P<0.01) respectively. Following GBSP treatment, microvilli of SMMC-7721 cells appeared thinner and the number of spherical cells increased markedly. Most significantly, the apoptosis bodies were formed on and around the spherical cells treated with GBSP.

Conclusion: GBSP could potentially induce the apoptosis of SMMC-7721 cells.

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Figures

Figure 1
Figure 1
The UV spectrum adsorption curve of GBSP from 200 nm to 400 nm
Figure 2
Figure 2
The profile of Sepharose 4B gel filtration chromatography
Figure 3
Figure 3
Standard linear calibration curve of Sephadex G-200 chromatography
Figure 4
Figure 4
In the absence of GBSP treatment, the apoptosis ratio of SMMC-7721 cells was 3.84% ± 0.55%, the cells debris was 0.46% ± 0.12% and the percentage of G2-M cells was 17.01% ± 1.28%.
Figure 5
Figure 5
In the presence of GBSP treatment, the apoptosis ratio of SMMC-7721 cells was 9.13% ± 1.48%, the cells debris was 0.06% ± 0.06%, and the percentage of G2-M cells was 11.77% ± 1.50%.
Figure 6
Figure 6
The majority of SMMC-7721 cells were of shuttle shape without GBSP treatment (× 2500)
Figure 7
Figure 7
There were more spherical SMMC-7721 cells after GBSP treatment (× 2200). These cells shrunk and apoptosis bodies were formed on and around them.
See this image and copyright information in PMC

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