Dipartimento di Sanità Pubblica, Sezione di Fisica, University of Parma Via Volturno 39, Parma, Italy.
Cancer Cell Int. 2011 Aug 22;11:30. doi: 10.1186/1475-2867-11-30.
The synergic action of KHCO3 and D-ribose is tested on A72 and HTB-126 cell lines proliferation using K:D-Rib solution. Altered Na+/K+ ATPase expression and activity were shown in patients with cancer. Studies in human epithelial-derived malignancies indicate that K+ depletion also occurs, contributing to the increased intracellular Na+/K+ ratio 1. D-ribose transformed to piruvate, enters into the Krebs's cycle and has a key role on energetic metabolism. The up-regulation of glycolysis in tumor cells is already well known and it is the rationale of F18-FDG PET diagnostic technique. D-ribose is synthesized by the non-oxidative transketolase PPP reaction.
Results with different K:D-Rib concentrations show that MTT salt interferes with K:D-Rib solution and therefore this method is not reliable. The UV/VIS measurements show that K:D-Rib solutions reduce MTT salt to formazan in absence of cells. Cell proliferation has then been evaluated analysing the digital photos of the Giemsa stained cells with MCID™ software. At 5 mM K:D-Rib concentration, the cell growth arrests between 48 h and 72 h; in fact the cell number after 48 h is around the same with respect to the control after 72 h. In case of HTB-126 human cancer cells, the growth rate was valuated counting the splitting times during 48 days: control cells were split sixteen times while 5 mM treated cells eleven times. Most relevant, the clonogenic assay shows that nine colonies are formed in the control cells while only one is formed in the 5 mM and none in 10 mM treated cells.
The K:D-Rib solution has an antioxidant behaviour also at low concentrations. Incubation with 5 mM K:D-Rib solution on A72 cells shows a cytostatic effect at 5 mM, but it needs more than 24 h of incubation time to evidence this effect on cell proliferation. At the same concentration on human HTB-126 cells, K:D-Rib solution shows a clear replication slowing but the cytostatic effect at 10 mM K:D-Rib solution only. Results on A72 cells indicate the K+ uptake could be determinant either to arrest or to slow down cell growth.
使用 K:D-Rib 溶液测试了 KHCO3 和 D-核糖对 A72 和 HTB-126 细胞系增殖的协同作用。已经表明,癌症患者的 Na+/K+ATP 酶表达和活性发生改变。在人类上皮源性恶性肿瘤的研究中,也发现钾耗竭,导致细胞内 Na+/K+ 比值增加 1。D-核糖转化为丙酮酸,进入克雷布斯循环,在能量代谢中具有关键作用。肿瘤细胞中糖酵解的上调已经众所周知,这也是 F18-FDG PET 诊断技术的基础。D-核糖是由非氧化转酮醇 PPP 反应合成的。
不同 K:D-Rib 浓度的结果表明,MTT 盐会干扰 K:D-Rib 溶液,因此这种方法不可靠。紫外/可见测量表明,在没有细胞的情况下,K:D-Rib 溶液将 MTT 盐还原为甲臜。然后通过使用 MCID™软件分析 Giemsa 染色细胞的数字照片来评估细胞增殖。在 5mM K:D-Rib 浓度下,细胞生长在 48 小时至 72 小时之间停滞;事实上,与 72 小时的对照相比,48 小时后的细胞数量相同。在 HTB-126 人类癌细胞的情况下,通过在 48 天内计算分裂次数来评估生长速率:对照细胞分裂 16 次,而 5mM 处理细胞分裂 11 次。最相关的是,集落形成实验表明,在对照细胞中形成了 9 个集落,而在 5mM 和 10mM 处理的细胞中分别形成了 1 个和无集落。
即使在低浓度下,K:D-Rib 溶液也具有抗氧化作用。在 A72 细胞中孵育 5mM K:D-Rib 溶液会显示出细胞生长抑制作用,但需要超过 24 小时的孵育时间才能在细胞增殖方面显示出这种作用。在相同浓度的 HTB-126 细胞上,K:D-Rib 溶液显示出明显的复制减缓,但只有在 10mM K:D-Rib 溶液中才显示出细胞生长抑制作用。A72 细胞的结果表明,钾的摄取可能是决定细胞生长停滞或减缓的决定因素。