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Soyasapogenol B exhibits anti-growth and anti-metastatic activities in clear cell renal cell carcinoma

  • Luping Wang
  • Junyu Wang
  • Hong Zhao
  • Guoping Jiang
  • Xiaojie Feng
  • Wenxia Sui
  • Hongling LiuEmail author
Original Article

Abstract

Clear cell renal cell carcinoma (ccRCC) is the most common type of human malignancies of the urological system. Soyasapogenol B (Soy B), an ingredient of soybean, has been found to exert anti-proliferative activities in vitro in human breast cancer cells. Our current study aimed to evaluate the effectiveness of Soy B against ccRCC. The effect of Soy B on cell viability was assessed by Cell Counting Kit-8 (CCK-8) assay. The effect of Soy B on cell proliferation was determined by colony formation assay. Apoptotic percentage was determined by flow cytometry following annexin V-FITC/propidium iodide (PI) double staining. JC-1 staining was performed to examine the change in mitochondrial membrane potential. Western blotting was used to determine the level of relevant proteins. Isobaric tags for relative and absolute quantification (iTRAQ) was then performed to identify the potential targets of Soy B. Quantitative real-time PCR (qRT-PCR) was performed to determine the mRNA level of sphingosine kinase 1 (SphK1). The SphK1 expression in ccRCC tissue from patients was examined by immunohistochemistry (IHC) assay. To validate the role of SphK1 involved in the pro-apoptotic activities of Soy B, overexpressed SphK1 vectors and shRNA targeting of SphK1 were utilized to transfected ccRCC cells. Moreover, a ccRCC xenograft murine model was used to analyze the therapeutic efficacy of Soy B in vivo. Soy B incubation led to a decrease in the number of viable cells in ccRCC cell lines and primary ccRCC cells. Soy B also suppressed the proliferation of two model ccRCC cell lines. Soy B promoted apoptotic cell death in a caspase-dependent manner. Moreover, our results showed that both extrinsic and intrinsic apoptotic signaling pathways were involved in Soy B-induced apoptosis. ITRAQ analysis identified SphK1 as most profoundly altered after the treatment of Soy B in ACHN cells. The mediatory role of SphK1 was validated when the pro-apoptotic activity of Soy B was significantly blocked by SphK1 overexpression, while SphK1 knockdown sensitized the ccRCC cells to Soy B. Moreover, in vivo studies also showed that Soy B could exhibit anti-cancer activities against ccRCC. Soy B triggers apoptotic cell death in vitro and in vivo in ccRCC by down-regulating SphK1. Our results highlight the possibility of using Soy B as a chemotherapeutic agent in the prevention and treatment of ccRCC.

Keywords

Soy B ccRCC SphK1 Anti-cancer 

Notes

Author contributions

Hongling Liu designed the research. Luping Wang, Junyu Wang, Hong Zhao, and Guoping Jiang conducted the experiments. Xiaojie Feng and Wenxia Sui analyzed the data. Luping Wang wrote the manuscript. All authors read and approved the manuscript.

Compliance with ethical standards

The study protocol was reviewed and approved by the Medical Ethics Committee of Qingdao Central Hospital, and all patients (or their guardians) provided written consent.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

210_2018_1607_Fig9_ESM.png (215 kb)
Supplementary Figure 1

HK-2 cells were treated with Soy B at indicated concentration for 24 or 48 h. Soy B treatment at 10 μM for 48 h significantly decreased cell viability. **P < 0.01. (PNG 215 kb)

210_2018_1607_MOESM1_ESM.tif (353 kb)
High resolution image (TIF 352 kb)
210_2018_1607_Fig10_ESM.png (793 kb)
Supplementary Figure 2

Soy B did not alter the expression of mitochondrial and cytosolic AIF and Endo G. (PNG 792 kb)

210_2018_1607_MOESM2_ESM.tif (1.1 mb)
High resolution image (TIF 1090 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Luping Wang
    • 1
  • Junyu Wang
    • 2
  • Hong Zhao
    • 2
  • Guoping Jiang
    • 1
  • Xiaojie Feng
    • 1
  • Wenxia Sui
    • 1
  • Hongling Liu
    • 1
    Email author
  1. 1.The Affiliated Hospital of Qingdao UniversityQingdaoChina
  2. 2.Qingdao Central HospitalQingdaoChina

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