Baqlouq Laila, Zihlif Malek, Hammad Hana, Thaib Tuqa M Abu
Department of Biology, Faculty of Science, The University of Jordan, Amman 11942, Jordan.
Department of Pharmacology, Faculty of Medicine, The University of Jordan, Amman 11942, Jordan.
Curr Mol Pharmacol. 2021;14(1):52-59. doi: 10.2174/1874467213666200521081653.
This study aims to identify the changes in the expression of hypoxia-inducible genes in seven different cancer cell lines that vary in their oxygen levels in an attempt to identify hypoxia biomarkers that can be targeted in therapy. Profiling of hypoxia inducible-gene expression of these different cancer cell lines can be used as baseline data for further studies.
Human cancer cell lines obtained from the American Type Culture Collection were used; MCF7 breast cancer cells, PANC-1 pancreatic cancer cells, PC-3 prostate cancer cells, SHSY5Y neuroblastoma brain cancer cells, A549 lung cancer cells, and HEPG2 hepatocellular carcinoma. In addition, we used the MCF10A non-tumorigenic human breast epithelial cell line as a normal cell line. The differences in gene expression were examined using real-time PCR array (PAHS- 032Z, Human Hypoxia Signaling Pathway PCR Array) and analyzed using the ΔΔCt method.
Almost all hypoxia-inducible genes showed a PO2-dependent up- and down-regulated expression. Noticeable gene expression differences were identified. The most important changes occurred in the HIF1α and NF-KB signaling pathways targeted genes and in central carbon metabolism pathway genes such as HKs, PFKL, and solute transporters.
This study identified possible hypoxia biomarkers genes such as NF-KB, HIF1α, HK, PFKL, and PIM1 that were expressed in all hypoxic cells. Pleiotropic pathways that play a role in inducing hypoxia directly, such as HIF1 α and NF-kB pathways, were upregulated. In addition, genes expressed only in the severe hypoxic liver and pancreatic cells indicate that severe and intermediate hypoxic cancer cells vary in their gene expression. Gene expression differences between cancer and normal cells showed the shift in gene expression profile to survive and proliferate under hypoxia.
本研究旨在确定七种不同癌细胞系中缺氧诱导基因表达的变化，这些细胞系的氧水平各不相同，以试图识别可在治疗中作为靶点的缺氧生物标志物。对这些不同癌细胞系的缺氧诱导基因表达进行分析可作为进一步研究的基线数据。
使用从美国典型培养物保藏中心获得的人癌细胞系；MCF7乳腺癌细胞、PANC-1胰腺癌细胞、PC-3前列腺癌细胞、SHSY5Y神经母细胞瘤脑癌细胞、A549肺癌细胞和HEPG2肝细胞癌。此外，我们使用MCF10A非致瘤性人乳腺上皮细胞系作为正常细胞系。使用实时PCR阵列（PAHS-032Z，人缺氧信号通路PCR阵列）检测基因表达差异，并使用ΔΔCt方法进行分析。
几乎所有缺氧诱导基因均呈现出依赖于氧分压的上调和下调表达。发现了明显的基因表达差异。最重要的变化发生在HIF1α和NF-κB信号通路的靶向基因以及诸如己糖激酶（HKs）、磷酸果糖激酶L（PFKL）和溶质转运蛋白等中心碳代谢途径基因中。
本研究确定了可能的缺氧生物标志物基因，如NF-κB、HIF1α、HK、PFKL和PIM1，这些基因在所有缺氧细胞中均有表达。直接参与诱导缺氧的多效性信号通路，如HIF1α和NF-κB信号通路，被上调。此外，仅在严重缺氧的肝脏和胰腺细胞中表达的基因表明，严重缺氧和中度缺氧的癌细胞在基因表达上存在差异。癌症细胞与正常细胞之间的基因表达差异显示了基因表达谱在缺氧条件下为存活和增殖而发生的转变。
