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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (12): 1174-1180.doi: 10.3736/jcim20091216

• Original Experimental Research • Previous Articles     Next Articles

Effects of Astragalus injection and its ingredients on proliferation and Akt phosphorylation of breast cancer cell lines

Ying Deng , Hong-feng Chen   

  1. Department of Breast Surgery, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
  • Received:2009-09-04 Accepted:2009-10-29 Online:2009-12-20 Published:2009-12-15
  • Contact: Hong-feng Chen E-mail:chhfluk@yahoo.com.cn

Objective

To observe the effects of Astragalus injection, astragaloside Ⅳ and formononetin on proliferation and Akt phosphorylation of basal-like human breast carcinoma cell lines MDA-MB-468 and MDA-MB-231.
Methods

The effects of different concentrations of Astragalus injection, astragaloside Ⅳ and formononetin on proliferation of breast cancer cell lines were assayed by methyl thiazolyl tetrazolium (MTT) assay, and their effects on phospho-Akt were assayed by in-cell Western blot method.
Results

The results of the MTT assay showed that the best concentrations of Astragalus injection, astragaloside Ⅳ, formononetin and astragaloside Ⅳ plus formononetin were 1 g/mL, 80 μg/mL, 40 μg/mL and 10 μg/mL plus 40 μg/mL respectively. After 1- or 2-day culture, Astragalus injection, astragaloside Ⅳ, formononetin and astragaloside Ⅳ plus formononetin decreased the expressions of p-Akt (Thr 308) and p-Akt (Ser 473) in MDA-MB-468 cells. Formononetin and astragaloside Ⅳ plus formononetin down-regulated the expression of p-Akt (Thr 308) protein in MDA-MB-231 cells after 1- and 2-day culture, but had no effects on the expression of p-Akt (Ser 473) protein in MDA-MB-231 cells.
Conclusion

Astragalus injection, astragaloside Ⅳ and formononetin can inhibit proliferation of breast cancer cell lines MDA-MB-468 and MDA-MB-231, and the antiproliferation effects vary according to their concentrations. And the antiproliferation mechanisms may be related to their down-regulation effects on Akt phosphorylation.

Key words: Astragalus injection, Astragaloside F, Formononetin, Breast cancer, Cell proliferation, Phospho-Akt

Table 1

Effects of Astragalus injection, astragaloside Ⅳ and formononetin on proliferation of breast cancer cell lines ($\bar{x}$±s)"

Group n OD value
MDA-MB-231 MDA-MB-468
Control 5 0.87±0.01 0.43±0.02
1 g/mL Astragalus injection 5 0.57±0.10*□ 0.28±0.02*□
0.1 g/mL Astragalus injection 5 0.96±0.02 0.46±0.05
40 μg/mL formononetin 5 0.37±0.05*△□ 0.15±0.02*△□
20 μg/mL formononetin 5 0.60±0.09*□ 0.15±0.01*△□
10 μg/mL formononetin 5 0.65±0.06*□ 0.20±0.01*△□
80 μg/mL astragaloside Ⅳ 5 0.71±0.06*△▲ 0.33±0.10*△▲
40 μg/mL astragaloside Ⅳ 5 0.73±0.03*△▲ 0.32±0.01*△▲
20 μg/mL astragaloside Ⅳ 5 0.86±0.09 0.40±0.01

Table 2

Effects of astragaloside Ⅳ combined with formononetin on proliferation of breast cancer cell lines ($\bar{x}$±s)"

Group n OD value
MDA-MB-231 MDA-MB-468
Control 5 0.87±0.01 0.43±0.02
1 g/mL Astragalus injection 5 0.59±0.05 0.28±0.02
40 μg/mL formononetin 5 0.38±0.05 0.15±0.02
80 μg/mL astragaloside Ⅳ 5 0.71±0.06 0.35±0.05
Astragaloside Ⅳ︰formononetin
1︰1 5 0.33±0.03*△□ 0.16±0.03*△□
1︰2 5 0.42±0.08*△□ 0.13±0.07*△□
1︰4 5 0.25±0.08*△▲□ 0.11±0.06*△□
2︰1 5 0.35±0.02*△□ 0.24±0.04*□
4︰1 5 0.56±0.05*□ 0.30±0.05*
8︰1 5 0.70±0.07 0.32±0.09*

"

Group n p-Akt Thr308 (Fluorescence value) p-Akt Ser473 (Fluorescence value)
1 d 2 d 1 d 2 d
Control 5 3.17±0.31 6.28±0.51 1.43±0.27 2.13±0.36
Astragalus injection 5 2.29±0.29* 4.64±0.96* 1.42±0.26 2.07±0.10
Formononetin 5 2.35±0.46* 3.74±0.39*△ 1.08±0.10* 1.81±0.25*
Astragaloside Ⅳ 5 2.39±0.41* 4.62±0.80* 1.27±0.22* 2.28±0.31
Astragaloside Ⅳ plus formononetin 5 2.52±0.45* 4.00±0.20*△ 1.11±0.09* 1.67±0.14*

Figure 1

Phospho-Akt expressions in MDA-MB-468 cells after 2-day exposure to different interventions A: p-Akt (Thr 308) level; B: p-Akt (Ser 473) level. C: Control group; AI: Astragalus injection group; F: Formononetin group; AⅣ: Astragaloside Ⅳ group; AⅣ+F: Astragaloside Ⅳ plus formononetin group."

"

Group n p-Akt Thr308 (Fluorescence value) p-Akt Ser473 (Fluorescence value)
1 d 2 d 1 d 2 d
Control 5 4.46±0.75 3.95±0.44 2.80±0.48 2.75±0.40
Astragalus injection 5 3.86±0.68 3.10±0.41* 2.68±0.45 2.20±0.27*
Formononetin 5 3.19±0.37* 3.05±0.33* 2.48±0.48 2.01±0.04*
Astragaloside Ⅳ 5 4.27±0.66 3.70±0.45* 2.44±0.44 2.40±0.28*
Astragaloside Ⅳ plus formononetin 5 3.67±0.54* 2.58±0.19*▲ 2.64±0.39 2.10±0.08*

Figure 2

Phospho-Akt expression in MDA-MB-231 cells after 2-day exposure to different interventions A: p-Akt (Thr 308) level; B: p-Akt (Ser 473) level. C: Control group; AI: Astragalus injection group; F: Formononetin group; AⅣ: Astragaloside Ⅳ group; AⅣ+F: Astragaloside Ⅳ plus formononetin group."

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