JS-K, a nitric oxide-releasing prodrug, induces breast cancer cell death while sparing normal mammary epithelial cells

Abstract

Abstract. Targeted therapy with reduced side effects is a major goal in cancer research. We investigated the effects of JS-K, a nitric oxide (NO) prodrug designed to release high levels of NO when suitably activated, on human breast cancer cell lines, on non-transformed human MCF-10A mammary cells, and on normal human mammary epithelial cells (HMECs). Cell viability assay, flow cytometry, electron microscopy, and Western blot analysis were used to study the effects of JS-K on breast cancer and on mammary epithelial cells. After a 3-day incubation, the IC 50 s of JS-K against the breast cancer cells ranged from 0.8 to 3 μM. However, JS-K decreased the viability of the MCF-10A cells by only 20% at 10-μM concentration, and HMECs were unaffected by 10 μM JS-K. Flow cytometry indicated that JS-K increased the percentages of breast cancer cells under-going apoptosis. Interestingly, flow cytometry indicated that JS-K increased acidic vesicle organelle formation in breast cancer cells, suggesting that JS-K induced autophagy in breast cancer cells. Electron microscopy confirmed that JS-K-treated breast cancer cells underwent autophagic cell death. Western blot analysis showed that JS-K induced the expression of microtubule light chain 3-II, another autophagy marker, in breast cancer cells. However, JS-K did not induce apoptosis or autophagy in normal human mammary epithelial cells. These data indicate that JS-K selectively induces programmed cell death in breast cancer cells while sparing normal mammary epithelial cells under the same conditions. The selective antitumor activity of JS-K warrants its further investigation in breast tumors.

Chemotherapeutic potential of diazeniumdiolate-based aspirin prodrugs in breast cancer

“…While the cytotoxicity of both aspirin and ionic diazeniumdiolates in various cancer cells lines is low (IC 50 > 1 mM) [72, 83, 97], NO-NSAIDs and related constructs significantly inhibit growth of cultured cancer cells as well as xenografts in mouse models [13, 26, 30, 64, 97–102]. Keefer and colleagues have previously shown that the derivatized diazeniumdiolate JS-K was significantly cytotoxic toward a variety of breast cancer cell lines (>50% reduction in viability at 1 μM JS-K) [102].…”

“…Keefer and colleagues have previously shown that the derivatized diazeniumdiolate JS-K was significantly cytotoxic toward a variety of breast cancer cell lines (>50% reduction in viability at 1 μM JS-K) [102]. The impact was more modest for non-malignant epithelial cells up to 10 μM JS-K. We have also previously observed that IPA/NO-aspirin and DEA/NO-aspirin selectively reduced proliferation of lung carcinoma cells (IC 50 values of ~100 μM) compared to normal endothelial cells [26].…”

Gasotransmitters in cancer: from pathophysiology to experimental therapy

“…There are several approaches to selectively directing NO to tumour cells. One involves exploiting the specific metabolic activity of cancer cells by using glutathione- S -transferase (GST)-activated NO donors, such as para-aminobenzoic acid (PABA)–NO and JS-K 46,47 — in some cases, in combination with multi-arm polymeric nanocarriers. 48,49 Another strategy is to use photoactivated NO donors, in some cases applied as supramolecular complexes.…”

Systemic RALA/iNOS Nanoparticles: A Potent Gene Therapy for Metastatic Breast Cancer Coupled as a Biomarker of Treatment

“…Many such donors exist and are broadly represented by the organic nitrates, metal-NO complexes, S -nitrosothiols, sydnonimines, diazeniumdiolates (NONOates), and ·NO-drug hybrids 4 . One ·NO-donating prodrug that has received particular attention is JS-K. JS-K induced apoptosis in a range of breast cancer cell lines but spared normal human microvascular endothelial cells (HMECs) and MCF-10A 7 . JS-K was recruited into the National Cancer Institute’s Rapid Access to Interventional Development (RAID) program, accelerating its progression as a potential therapeutic agent 8 …”