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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (8): 741-745.doi: 10.3736/jcim20090807

• Original Experimental Research • Previous Articles     Next Articles

Effects of Feiyanning Decoction on proliferation of lung adenocarcinoma cell line and their production of interleukin-6 and interleukin-8 induced by tumor necrosis factor-α

Ju-yong Wang,Zhan Zheng, Zhen-ye Xu, Jing Guo, Wan So   

  1. Cancer Institute of Traditional Chinese Medicine,Longhua Hospital,Shanghai University of Traditional Chinese Medicine, Shanghai 200032,China
  • Received:2009-05-04 Accepted:2009-07-08 Online:2009-08-20 Published:2009-08-15
  • Contact: Ju-yong Wang E-mail:wangjuyong1@126.com

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Objective

To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on proliferation of lung adenocarcinoma cell line A549 cells and their production of interleukin-6 (IL-6) and IL-8 induced by tumor necrosis factor-α (TNF-α).

Methods

A549 cells were incubated with rat serum containing Feiyanning Decoction at 15% for 24, 48 and 72 h respectively. The cell proliferation was examined by 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium, monosodium salt assay (WST-8). The production of IL-6 and IL-8 was tested by enzyme-linked immunosorbent assay after 48-hour treatment of reagents, and the expressions of IL-6 and IL-8 mRNAs were detected by reverse transcription-polymerase chain reaction. 

Results

Serum containing Feiyanning Decoction had obvious inhibitive functions in A549 cell proliferation after 48- and 72-treatment. TNF-α (1 μg/L) strongly induced the production of IL-6 and IL-8 as compared with the control serum in A549 cells, and the induced cytokine production was significantly suppressed by 15% serum containing Feiyanning Decoction (P<0.01). In addition, serum containing Feiyanning Decoction could inhibit the mRNA expressions of IL-6 and IL-8 (P<0.01).
Conclusion

Feiyanning Decoction can inhibit IL-6 and IL-8 production induced by TNF-α. It is therefore expected to be a new strategy for treating lung cancer.

Key words: Feiyanning Decoction, Cytokine, Lung neoplasms, Serologic pharmacology, In vitro

"

Gene Primer sequence Size of PCR product (bp)
IL-6 Forward: 5'-GATGCTACCAAACTGGATATAATC-3' 269
Reverse: 5'-GGTCCTTAGCCACTCCTTCTGTG-3'
IL-8 Forward: 5'-CTGTGTGAAGCTGCAGTTCT-3' 180
Reverse: 5'-TAGGCAGACCTCGTTTCCAT-3'
β-actin Forward: 5'-AAGTACTCCGTGTGGATCGG-3' 547
Reverse: 5'-TCAAGTTGGGGGACAAAAAG-3'

"

Group n OD value Rate of inhibition after treatment (%)
24 h 48 h 72 h 24 h 48 h 72 h
Control serum 4 0.325±0.014 0.497±0.021 0.521±0.019 0 0 0
Serum containing Feiyanning
Decoction		 4 0.344±0.028 0.359±0.020 0.321±0.022 0.05±0.62 30.85±1.62* 37.66±1.48**

Figure 1

Effects of serum containing Feiyanning Decoction on IL-6 and IL-8 levels induced by TNF-α in A549 cells Confluent cells were incubated for 48 h with or without 15% serum containing Feiyanning Decoction, or TNF-α (1 μg/L). Concentrations of IL-6 and IL-8 in supernatant were measured by ELISA. Bars represent $\bar{x}$±s (n=3). **P<0.01, vs TNF-α."

Figure 2

Effects of serum containing Feiyanning Decoction on IL-6 and IL-8 mRNA expressions in A549 cells After the treatment, RNA was extracted. Levels of cytokines and β-actin mRNAs were examined by RT-PCR. The three panels on left show the expression levels of IL-6, IL-8 and β-actin mRNAs; bar graphs at the right show the expression levels of IL-6 or IL-8 relative to those of β-actin. The experiments were performed in triplicate. The data were represented as $\bar{x}$±s (n=3). **P<0.01, vs TNF-α."

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