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Apoptotic signaling induced by benzamide riboside: an in vitro study

Sujata Pathak1, Chandresh Sharma, H N Jayaram

  • 1Department of Biochemistry, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.

Molecular and Cellular Biochemistry
|March 6, 2009
PubMed

Insights

Benzamide riboside (BR) effectively induces apoptosis in human cancer cells. This novel anticancer agent triggers the intrinsic mitochondrial pathway, offering a new therapeutic strategy for cancer treatment.

Area of Science:

  • Oncology
  • Molecular Biology
  • Cell Biology

Background:

  • Benzamide riboside (BR) is recognized as a potent anticancer agent with significant cytotoxic effects on various malignant cell lines.
  • The precise molecular mechanisms underlying BR-induced apoptosis remain incompletely understood.

Purpose of the Study:

  • To investigate and clarify the specific apoptotic signaling pathways activated by Benzamide riboside (BR) in diverse human cancer cell lines.
  • To elucidate the role of key apoptotic proteins and cellular events in BR-mediated cell death.

Main Methods:

  • Morphological analysis and flow cytometry were employed to assess apoptosis induction in SiHa, Hep2, and Ca Ski cells treated with BR.
  • Western blotting and biochemical assays were used to evaluate the expression levels of apoptosis-related proteins (Bcl-2, Bcl-xL, Bax, p53, AIF) and the activity of caspases (caspase-3, caspase-9).
  • Mitochondrial integrity was assessed by monitoring cytochrome c release into the cytosol.

Main Results:

  • Benzamide riboside (BR) at 50 microM demonstrated significant apoptosis induction in SiHa, Hep2, and Ca Ski cells.
  • BR treatment led to the downregulation of anti-apoptotic proteins Bcl-2 and Bcl-xL, while Bax expression remained unchanged.
  • Increased p53 levels, elevated caspase-3 and -9 activities, PARP cleavage, and cytochrome c release from mitochondria were observed, indicating activation of the intrinsic apoptotic pathway.

Conclusions:

  • Benzamide riboside (BR) effectively induces apoptosis in SiHa, Hep2, and Ca Ski cancer cells.
  • The findings strongly suggest that BR activates apoptosis predominantly through the intrinsic mitochondrial pathway.
  • BR's mechanism involves modulating Bcl-2 family proteins, activating caspases, and releasing cytochrome c, highlighting its potential as an anticancer therapeutic.

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