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Induction of Apoptosis in Rat Peripheral Blood Lymphocytes by the Anticancer Drug CI-994 (Acetyldinaline)(*).

Graziano MJ, Spoon TA, Cockrell EA, Rowse PE, Gonzales AJ - J. Biomed. Biotechnol. (2001)

Bottom Line: Morphological and biochemical features of apoptosis were evaluated using several techniques, and lactate dehydrogenase (LDH) release was measured as an indicator of cell necrosis.After 24 hours, concentration-dependent increases in apoptosis characterized by DNA condensation, DNA fragmentation, and/or externalization of phosphatidyl serine were seen at CI-994 concentrations as low as 1 &mgr;M and were statistically significant beginning at 10 &mgr;M.Ultrastructural analysis confirmed the presence of DNA condensation, DNA fragmentation, cell shrinkage, and membrane blebbing in cells exposed to 30 &mgr;M CI-994.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
CI-994 (acetyldinaline) is an investigational anticancer drug currently in clinical trials. In preclinical safety studies in rats and dogs, CI-994 resulted in significant toxicity to bone marrow and lymphoid tissue. To determine if apoptosis was involved in CI-994 toxicity, peripheral blood lymphocytes were isolated from untreated male Wistar rats and exposed to CI-994 (1, 3, 10, or 30 &mgr;M) in vitro for up to 24 hours. Morphological and biochemical features of apoptosis were evaluated using several techniques, and lactate dehydrogenase (LDH) release was measured as an indicator of cell necrosis. No evidence of apoptosis or necrosis was detected in lymphocytes exposed to CI-994 for 4 hours. After 24 hours, concentration-dependent increases in apoptosis characterized by DNA condensation, DNA fragmentation, and/or externalization of phosphatidyl serine were seen at CI-994 concentrations as low as 1 &mgr;M and were statistically significant beginning at 10 &mgr;M. Ultrastructural analysis confirmed the presence of DNA condensation, DNA fragmentation, cell shrinkage, and membrane blebbing in cells exposed to 30 &mgr;M CI-994. After 24 hours, the percent of maximum LDH release from lymphocytes treated with 10 and 30 &mgr;M CI-994 was 7% and 15%, respectively, compared with 0% in the controls. In comparison, morphological changes of apoptosis detected by fluorescent microscopy were observed in 79% of the lymphocytes at these two concentrations. Additionally, apoptosis was seen in more than 24% of lymphocytes exposed to 1 and 3 &mgr;M CI-994, whereas maximum LDH release was less than or equal to 1% at these concentrations. These results show that apoptosis is the primary mode of cell death in rat lymphocytes exposed to CI-994 in vitro.

No MeSH data available.


Related in: MedlinePlus

Chemical structure of CI-994.
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Figure 1: Chemical structure of CI-994.

Mentions: CI-994 (acetyldinaline; Figure 1) is an investigational anticancer drug with activity in a broad spectrum of mouse, rat,and human tumor models [1, 2, 3, 4, 5]. CI-994 can also potentiate the activity of other standard chemotherapeuticagents. For example, simultaneous administration of CI-994 and gem-citabine results in greater than additive activity in a preclinical model of squamous cell lung carcinoma [6]. Although the mechanism of action of CI-994 is unknown, it appears to be novel when compared with other existing anticancer agents. CI-994 is not an antimetabolite, does not covalently bind to or intercalate DNA, and does not affect microtubule synthesis [7]. Studies have shown that CI-994 can inhibit a 16 kDa nuclear phosphoprotein and increase histone acetylation [8, 9]. Currently, CI-994 is in phase II clinical trials.


Induction of Apoptosis in Rat Peripheral Blood Lymphocytes by the Anticancer Drug CI-994 (Acetyldinaline)(*).

Graziano MJ, Spoon TA, Cockrell EA, Rowse PE, Gonzales AJ - J. Biomed. Biotechnol. (2001)

Chemical structure of CI-994.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC113775&req=5

Figure 1: Chemical structure of CI-994.
Mentions: CI-994 (acetyldinaline; Figure 1) is an investigational anticancer drug with activity in a broad spectrum of mouse, rat,and human tumor models [1, 2, 3, 4, 5]. CI-994 can also potentiate the activity of other standard chemotherapeuticagents. For example, simultaneous administration of CI-994 and gem-citabine results in greater than additive activity in a preclinical model of squamous cell lung carcinoma [6]. Although the mechanism of action of CI-994 is unknown, it appears to be novel when compared with other existing anticancer agents. CI-994 is not an antimetabolite, does not covalently bind to or intercalate DNA, and does not affect microtubule synthesis [7]. Studies have shown that CI-994 can inhibit a 16 kDa nuclear phosphoprotein and increase histone acetylation [8, 9]. Currently, CI-994 is in phase II clinical trials.

Bottom Line: Morphological and biochemical features of apoptosis were evaluated using several techniques, and lactate dehydrogenase (LDH) release was measured as an indicator of cell necrosis.After 24 hours, concentration-dependent increases in apoptosis characterized by DNA condensation, DNA fragmentation, and/or externalization of phosphatidyl serine were seen at CI-994 concentrations as low as 1 &mgr;M and were statistically significant beginning at 10 &mgr;M.Ultrastructural analysis confirmed the presence of DNA condensation, DNA fragmentation, cell shrinkage, and membrane blebbing in cells exposed to 30 &mgr;M CI-994.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
CI-994 (acetyldinaline) is an investigational anticancer drug currently in clinical trials. In preclinical safety studies in rats and dogs, CI-994 resulted in significant toxicity to bone marrow and lymphoid tissue. To determine if apoptosis was involved in CI-994 toxicity, peripheral blood lymphocytes were isolated from untreated male Wistar rats and exposed to CI-994 (1, 3, 10, or 30 &mgr;M) in vitro for up to 24 hours. Morphological and biochemical features of apoptosis were evaluated using several techniques, and lactate dehydrogenase (LDH) release was measured as an indicator of cell necrosis. No evidence of apoptosis or necrosis was detected in lymphocytes exposed to CI-994 for 4 hours. After 24 hours, concentration-dependent increases in apoptosis characterized by DNA condensation, DNA fragmentation, and/or externalization of phosphatidyl serine were seen at CI-994 concentrations as low as 1 &mgr;M and were statistically significant beginning at 10 &mgr;M. Ultrastructural analysis confirmed the presence of DNA condensation, DNA fragmentation, cell shrinkage, and membrane blebbing in cells exposed to 30 &mgr;M CI-994. After 24 hours, the percent of maximum LDH release from lymphocytes treated with 10 and 30 &mgr;M CI-994 was 7% and 15%, respectively, compared with 0% in the controls. In comparison, morphological changes of apoptosis detected by fluorescent microscopy were observed in 79% of the lymphocytes at these two concentrations. Additionally, apoptosis was seen in more than 24% of lymphocytes exposed to 1 and 3 &mgr;M CI-994, whereas maximum LDH release was less than or equal to 1% at these concentrations. These results show that apoptosis is the primary mode of cell death in rat lymphocytes exposed to CI-994 in vitro.

No MeSH data available.


Related in: MedlinePlus