1Department of Medical Biology and Genetics, Faculty of Medicine, Gazi University, Besevler, Ankara, Turkey.
Hum Exp Toxicol. 2014 Apr;33(4):414-23. doi: 10.1177/0960327113499050. Epub 2013 Aug 5.
Statins induce antiproliferative effects and apoptotic response in various cancer cell types. Moreover, they also sensitize tumor cell lines from different origins to many agents. We aimed to investigate possible effects of Mevastatin (Mev) alone and sequential treatment of 5'-aza-2-deoxycitidine (DAC) and Mev on HL-60 cell line using XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) assay, lactate dehydrogenase release assay, flourescence microscopy, DNA fragmentation analysis, determination of DNA synthesis rate, and active caspase-3 assay. Messenger RNA (mRNA) expression of apoptotic and antiapoptotic genes were also evaluated by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) for BAX, BCL2, and XIAP genes and quantitative Real-time PCR for CASP3, CASP8, and CASP9 genes. We showed that treatment with Mev alone and DAC followed by Mev resulted in apoptotic response in a time- and dose-dependent manner. We also found that pretreatment with DAC sensitized HL-60 cells to Mev and caused more apoptotic cell death than Mev-alone treatment via caspase-3 activation and DNA fragmentation. Moreover, sequential addition of Mev after DAC diminished DNA synthesis rate more effectively than Mev-alone treatment. Furthermore, DAC pretreatment significantly increased CASP3 and CASP9 mRNA expression even with lower doses of Mev. BAX, BCL2, and XIAP gene mRNA levels were also found to be changed in the presence of DAC and Mev. Determination of the exact molecular effects of statins and DAC would allow us to identify new molecular targets to develop more effective treatment regimens for cancer.
他汀类药物在多种癌细胞类型中诱导抗增殖作用和凋亡反应。此外，它们还能使来自不同来源的肿瘤细胞系对许多药物敏感。我们旨在使用 XTT（2,3-双-（2-甲氧基-4-硝基-5-磺苯基）-2H-四唑-5-羧基苯胺）测定法、乳酸脱氢酶释放测定法、荧光显微镜检查、DNA 片段化分析、DNA 合成率测定和活性半胱天冬酶-3 测定法，研究单独使用美伐他汀（Mev）以及顺序使用 5'-氮杂-2'-脱氧胞苷（DAC）和 Mev 对 HL-60 细胞系的可能影响。还通过半定量逆转录聚合酶链反应（RT-PCR）评估凋亡和抗凋亡基因的信使 RNA（mRNA）表达，用于 BAX、BCL2 和 XIAP 基因，以及定量实时 PCR 用于 CASP3、CASP8 和 CASP9 基因。结果表明，单独使用 Mev 以及 DAC 继之以 Mev 处理可导致时间和剂量依赖性的凋亡反应。还发现，DAC 预处理可使 HL-60 细胞对 Mev 敏感，并通过 caspase-3 激活和 DNA 片段化导致比单独使用 Mev 更多的凋亡细胞死亡。此外，与单独使用 Mev 相比，顺序添加 Mev 后可更有效地降低 DNA 合成率。此外，即使使用较低剂量的 Mev，DAC 预处理也可显著增加 CASP3 和 CASP9 mRNA 的表达。还发现 BAX、BCL2 和 XIAP 基因的 mRNA 水平在 DAC 和 Mev 的存在下发生了变化。确定他汀类药物和 DAC 的确切分子作用将使我们能够确定新的分子靶点，以开发更有效的癌症治疗方案。
