Human MCF-7 breast cancer cells were exposed to a Random Positioning Machine (RPM). After 24 hours (h) the cells grew either adherently within a monolayer (AD) or within multicellular spheroids (MCS). AD and MCS populations were separately harvested, their cellular differences were determined performing qPCR on genes, which were differently expressed in AD and MCS cells. Gene array technology was applied to detect RPM-sensitive genes in MCF-7 cells after 24 h. Furthermore, the capability to form multicellular spheroids in vitro was compared with the intracellular distribution of NF-kappaB (NFκB) p65. NFκB was equally distributed in static control cells, but predominantly localized in the cytoplasm in AD cells and nucleus in MCS cells exposed to the RPM. Gene array analyses revealed a more than 2-fold change of only 23 genes including some whose products are affected by oxygen levels or regulate glycolysis. Significant upregulations of the mRNAs of enzymes degrading heme, of ANXA1, ANXA2, CTGF, CAV2 and ICAM1, as well as of FAS, Casp8, BAX, p53, CYC1 and PARP1 were observed in MCS cells as compared with 1g-control and AD cells. An interaction analysis of 47 investigated genes suggested that HMOX-1 and NFκB variants are activated, when multicellular spheroids are formed.

中文翻译：

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NFκB在随机定位机上培养的人乳腺癌细胞球状形成中的作用。

将人MCF-7乳腺癌细胞暴露于随机定位仪（RPM）。24小时（h）后，细胞在单层（AD）或多细胞球体（MCS）内粘附生长。分别收获AD和MCS群体，通过对基因进行qPCR确定它们的细胞差异，所述基因在AD和MCS细胞中表达不同。24小时后，应用基因阵列技术检测MCF-7细胞中RPM敏感基因。此外，将体外形成多细胞球体的能力与NF-κB（NFκB）p65的细胞内分布进行了比较。NFκB平均分布在静态对照细胞中，但主要位于暴露于RPM的AD细胞的细胞质和MCS细胞的细胞核中。基因阵列分析显示，只有23个基因发生了超过2倍的变化，包括一些其产物受氧水平或调节糖酵解影响的基因。与1g对照和AD细胞相比，在MCS细胞中观察到降解血红素的酶，ANXA1，ANXA2，CTGF，CAV2和ICAM1以及FAS，Casp8，BAX，p53，CYC1和PARP1的mRNA显着上调。 。对47个研究基因的相互作用分析表明，当形成多细胞球体时，HMOX-1和NFκB变体被激活。