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Tounongsan (透脓散) extract induces apoptosis in cultured Raji cells

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Abstract

Objectiv

To investigate the effects of Tounongsan (透脓散) extract (TNSE) on proliferation and e: apoptosis of the human lymphoma cell line Raji and its possible mechanism of action.

Methods

The viability of: TNSE-treated Raji cells was measured by a 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell apoptosis was determined by flow cytometry. The molecular mechanisms of TNSE-mediated apoptosis were further investigated by reverse transcription-polymerase chain reaction (RT-PCR) analysis of the mRNA expression of nuclear factor κB (NF-κB), Bcl-xL, Bcl-2-associated death promoter (Bad), caspase-9 and caspase-3. Western blotting was used to detect the protein expressions of NF-κB, Bad, cleaved caspase-9 and cleaved caspase-3.

Results

TNSE inhibited Raji cell proliferation in dose- and time-dependent manners.: After 48-h treatment with various concentrations of TNSE (125, 250 and 500 μg/mL), the apoptosis rates of Raji cell were 12.23%±1.98% (P<0.05), 20.97%±3.96% (P<0.01) and 30.4%±4.87% ( P<0.01), respectively, compared with those of the control (6.02%±1.01%). RT-PCR demonstrated that NF-κB mRNA expression was significantly downregulated in Raji cells treated with 250 μg/mL TNSE for 48 h (P<0.05), while Bad, caspase-9 and caspase-3 mRNA levels were upregulated (P<0.05). Moreover, TNSE treatment resulted in downregulation of NF-κB protein expression and strikingly upregulated protein expressions of Bad, cleaved caspase-9, cleaved caspase-3 in a dose-dependent manner, as determined by Western blot.

Conclusion

TNSE exhibits significant: anti-proliferative and apoptotic effects in Raji cells, which may be involved in regulation of NF-κB and Bad, and activation of caspase-9 and caspase-3.

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Authors and Affiliations

  1. First Clinical Medical College of Nanjing University of Chinese Medicine, Nanjing, 210046, China

    Liang-hua Fang  (房良华)

  2. Oncology Department of Jiangsu Provincial Hospital of Traditional Chinese Medicine, Nanjing, 210029, China

    Rui-ping Wang  (王瑞平), Shou-you Hu  (胡守友), Li Zhang  (张 力) & Shen-lin Liu  (刘沈林)

Authors
  1. Liang-hua Fang  (房良华)
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  2. Rui-ping Wang  (王瑞平)
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  3. Shou-you Hu  (胡守友)
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  4. Li Zhang  (张 力)
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  5. Shen-lin Liu  (刘沈林)
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Corresponding author

Correspondence to Shen-lin Liu  (刘沈林).

Additional information

Supported by A Project of the Priority Academic Program Development of Jiangsu Higher Education Institutions (No. 012062003010), Project of Top Talented Personnel in Six Profession In Jiangsu Province (No. 2011-WS-049) and Project of Jiangsu Province Hospital of Traditional Chinese Medicine (No. 2010-Y1008).

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Fang, Lh., Wang, Rp., Hu, Sy. et al. Tounongsan (透脓散) extract induces apoptosis in cultured Raji cells. Chin. J. Integr. Med. 18, 522–528 (2012). https://doi.org/10.1007/s11655-012-1145-4

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  • DOI: https://doi.org/10.1007/s11655-012-1145-4

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