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Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (1): 61-66.doi: 10.3736/jcim20100112

• Original Experimental Research • Previous Articles     Next Articles

Effects of cycloartocarpin A and artocarpin extracted from Fructus Artocarpi Heterophylli on apoptosis of SMMC-7721 and SGC-7901 cells

YL Yang,AJ Hou,HQ Zhang,H Shen,QS Li,CC Zhang,GF Zhu   

  • Received:2009-08-25 Accepted:2009-11-10 Online:2010-01-20 Published:2010-01-15
  • Contact: GF Zhu E-mail:zhuguofush@yahoo.com.cn

Objective

To investigate the effects of cycloartocarpin A (ACR-2) and artocarpin (ACR-3), monomeric compounds isolated from Fructus Artocarpi Heterophylli, on apoptosis of SMMC-7721 and SGC-7901 cell lines.
Methods

SMMC-7721 and SGC-7901 cells were routinely cultured, and divided into experiment group and control group. The SMMC-7721 cells were treated with different concentrations of ACR-2 (3.46×10 –3, 13.82×10 –3, 55.30×10 –3 mmol/L) and ACR-3 (6.88×10 –3, 27.52×10 –3, 110.09×10 –3 mmol/L), and the SGC-7901 cells were also treated with different concentrations of ACR-2 (8.06×10 –3, 32.26×10 –3, 129.03×10 –3 mmol/L) and ACR-3 (2.87×10 –3, 11.47×10 –3, 45.87×10 –3 mmol/L), with PBS (DMSO<0.1%) as control treatment. Cell apoptosis was measured by double labeled staining with Hoechst33342/propidium iodide (PI) and TdT-mediated dUTP-biotin nick end labeling (TUNEL) and flow cytometry.
Results

ACR-2 and ACR-3 could induce apoptosis of SMMC-7721 and SGC-7901 cells. Some of SMMC-7721 and SGC-7901 cells demonstrated typical apoptosis after being treated with ACR-2 and ACR-3. Hoechst33342/PI staining showed that cells were fraught with overlapping nuclei and nuclear debris or lobule, and the nuclear appeared light blue. TUNEL showed that cells permeated with overlapping nuclei and nuclear debris or lobule, and the nuclear appeared brown. Less apoptotic cells were observed in negative control group, and the nuclear appeared light blue. The apoptosis rates of SMMC-7721 and SGC-7901 cells in the ACR-2 and ACR-3 treated groups were significant higher than those in the control group (P<0.05, P<0.01).
Conclusion

ACR-2 and ACR-3 can induce apoptosis of SMMC-7721 and SGC-7901 cells.

Table 1

Concentrations of ACR-2 and ACR-3"

Cell n ACR-2 (mmol/L) ACR-3 (mmol/L)
SMMC-7721 cells 6 3.46×10–3 6.88×10–3
6 13.82×10–3 27.52×10–3
6 55.30×10–3 110.09×10–3
SGC-7901 cells 6 8.06×10–3 2.87×10–3
6 32.26×10–3 11.47×10–3
6 129.03×10–3 45.87×10–3

Table 2

Effects of ACR-2 and ACR-3 on apoptosis of SMMC-7721 cells after 48-hour treatment ($\bar{x}±s$, %)"

Group n Apoptosis rate
Fluorescence microscopy TUNEL method Flow cytometry
PBS (DMSO<0.1%) 6 0.35±0.02 0.51±0.06 0.40±0.01
3.46×10–3 mmol/L ACR-2 6 5.33±1.56* 5.59±1.27* 5.50±0.24**
13.82×10–3 mmol/L ACR-2 6 6.00±1.53* 8.76±1.23** 5.91±0.27**
55.30×10–3 mmol/L ACR-2 6 21.13±3.51** 22.20±2.15** 18.80±1.21**
6.88×10–3 mmol/L ACR-3 6 4.78±0.57** 10.56±2.78* 9.43±0.45**
27.52×10–3 mmol/L ACR-3 6 7.71±1.55* 8.02±1.63* 6.60±0.56**
110.09×10–3 mmol/L ACR-3 6 25.09±2.18** 27.05±2.63** 30.11±3.45**

Table 3

Effects of ACR-2 and ACR-3 on apoptosis of SGC-7901 cells after 48-hour treatment ($\bar{x}±s$, %)"

Group n Apoptosis rate
Fluorescence microscopy TUNEL method Flow cytometry
PBS (DMSO<0.1%) 6 0.38±0.04 0.23±0.08 0.20±0.02
8.06×10–3 mmol/L ACR-2 6 3.72±0.55** 7.37±1.13** 6.30±1.07*
32.26×10–3 mmol/L ACR-2 6 15.17±3.44* 18.49±2.63** 20.04±0.35**
129.03×10–3 mmol/L ACR-2 6 24.42±1.34** 27.92±1.21** 33.21±3.37**
2.87×10–3 mmol/L ACR-3 6 7.69±1.08** 13.17±1.99** 12.70±2.32*
11.47×10–3 mmol/L ACR-3 6 24.57±2.17** 19.96±3.60* 21.11±2.65**
45.87×10–3 mmol/L ACR-3 6 33.23±4.38** 34.78±1.42** 30.83±2.74**
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