Cytotoxic Activity and Inhibition of Tumor Cell Invasion by Derivatives of a Chemically Modified Tetracycline CMT-3 (COL-3)

Bal   L.   Lokeshwar; Eva      Escatel; Baoqian      Zhu

Abstract

Tetracyclines such as chlortetracycline and doxycycline with antimicrobial activity were reported to possess cytostatic and cytotoxic activity against mammalian tumor cells, often at high doses. Non-antimicrobial chemically modified tetracyclines (CMTs), with limited systemic toxicity but with significant tumor cell toxicity and anti-metastatic activity, are attractive for long term treatment for cancer. We recently reported one such CMT, 6-deoxy, 6-demethyl 4-dedimethylamino tetracycline (CMT-3) is a potent anti-tumor and anti-metastatic drug. Here we report on the anti-cell proliferation and anti-invasive activity of five nitro derivatives of CMT-3 (CMT-3N). All the five CMT-3Ns (CMT-302, CMT-303, CMT-306, CMT-308 and CMT-316) inhibited in vitro cell proliferation of prostate cancer cells. The 50 percent growth inhibition concentration (IC 50 ) of CMT-3Ns was similar to that of CMT-3. Although CMT-3 was by far the most potent anti-cell proliferation drug, all CMT-3Ns except CMT-303 and CMT-308 had similar anti-cell proliferation activity (IC 50 : 2.5 -5.7 micro g/ml). IC 50 s for CMT-303 and CMT-308 were - 8.1 and -12.4 micro g/ml, respectively. Activity against tumor cell invasion was tested in vitro using the Matrigel invasion assay. All CMT-3Ns had similar anti- invasive activity. While cytotoxic activity of CMT-3 was strongly associated with cell death-effector caspase activation, mitochondrial permeablization and apoptosis, the CMT-3Ns weakly induced apoptosis and did not activate Caspase-3. However, the CMT-3Ns were able to induce mitochondrial permeabilization. This dichotomous mechanism of cytotoxic activity of CMTs may have significance in their selection for clinical application.

Keywords: tumor cell invasio, modified tetracycluine, potent anti tumor, metastatic drugs, existing chemotherapeutics drugs, dedmethylamino tetracycline, culture conditioned medium, cell death assay, Chromatine fragmentation, phoshatidyl serine, capase activation, depolarization of mitochondria, cell proliferation, flow cytometry