Instituto de Inmunología, Facultad Ciencias Médicas, Universidad Nacional Rosario, Rosario, Argentina.
Immunopharmacol Immunotoxicol. 2013 Aug;35(4):478-86. doi: 10.3109/08923973.2013.811597.
We have previously reported that benznidazole (BZL), known for its trypanocidal action, has anti-proliferative activity against different cell lines like HeLa and Raw 264.7 among others. At the moment, it has not been reported if the anti-proliferative effect of BZL is similar for non-adherent hematopoietic cells like was reported for adherent cancer cell lines.
We aimed to investigate the efficacy of BZL on the growth of the leukemic cell lines THP-1 and OCI/AML3.
We evaluated cell proliferation by [³H]-thymidine incorporation and MTT reduction as well as cell death by lactate dehydrogenase (LDH) activity. We assessed apoptosis by flow cytometry for detection of annexin V-positive and propidium iodide-negative cells, along with nuclear morphology by diamidino-2-phenolindole (DAPI) staining. Western blot studies were performed to evaluate changes in cell cycle proteins in BZL-treated cells.
BZL significantly reduced proliferation of both cell lines without inducing cell death. Likewise it produced no significant differences in apoptosis between treated cells and controls. In addition, flow cytometry analysis indicated that BZL caused a larger number of THP-1 cells in G0/G1 phase and a smaller number of cells in S phase than controls. This was accompanied with an increase in the expression of the CDK inhibitor p27 and of cyclin D1, with no significant differences in the protein levels of CDK1, CDK2, CDK4, cyclins E, A and B as compared to controls.
BZL inhibits the proliferation of leukemic non-adherent cells by controlling cell cycle at G0/G1 cell phase through up-regulation of p27.
我们之前曾报道过，苯硝唑（BZL）具有杀锥虫作用，对包括 HeLa 和 Raw 264.7 在内的不同细胞系具有抗增殖活性。目前，尚未报道 BZL 的抗增殖作用是否与粘附癌细胞系类似，对非粘附造血细胞也具有相似的作用。
我们旨在研究 BZL 对白血病细胞系 THP-1 和 OCI/AML3 的生长的影响。
我们通过[³H]-胸苷掺入和 MTT 还原评估细胞增殖，通过乳酸脱氢酶（LDH）活性评估细胞死亡。通过流式细胞术检测 Annexin V 阳性和碘化丙啶阴性细胞来评估细胞凋亡，同时通过二脒基-2-苯并咪唑（DAPI）染色评估核形态。通过 Western blot 研究评估 BZL 处理细胞中环细胞蛋白的变化。
BZL 显著降低了两种细胞系的增殖，而没有诱导细胞死亡。同样，与对照组相比，BZL 处理的细胞和对照组之间的凋亡也没有显著差异。此外，流式细胞术分析表明，BZL 使 THP-1 细胞在 G0/G1 期的数量增加，而 S 期的细胞数量减少，而对照组则没有。这伴随着 CDK 抑制剂 p27 的表达增加，以及 cyclin D1 的表达增加，与对照组相比，CDK1、CDK2、CDK4、cyclins E、A 和 B 的蛋白水平没有显著差异。
BZL 通过上调 p27 控制细胞周期在 G0/G1 期，从而抑制非粘附白血病细胞的增殖。
