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碳酸氢钾和 D-核糖对体外 A72 犬和 HTB-126 人癌细胞系增殖的影响。

Potassium bicarbonate and D-ribose effects on A72 canine and HTB-126 human cancer cell line proliferation in vitro.

机构信息

Dipartimento di Sanità Pubblica, Sezione di Fisica, University of Parma Via Volturno 39, Parma, Italy.

出版信息

Cancer Cell Int. 2011 Aug 22;11:30. doi: 10.1186/1475-2867-11-30.

Abstract

BACKGROUND

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

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.

CONCLUSIONS

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.

摘要

背景

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。

结果

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。

结论

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1725/3173286/17d094d45448/1475-2867-11-30-1.jpg

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