Skrzycki Michał, Czeczot Hanna, Mielczarek-Puta Magdalena, Otto-Ślusarczyk Dagmara, Graboń Wojciech
a Department of Biochemistry , Medical University of Warsaw , Warsaw , Poland.
J Recept Signal Transduct Res. 2017 Jun;37(3):252-258. doi: 10.1080/10799893.2016.1217882. Epub 2016 Nov 9.
Tumor cells due to distance from capillary vessels exist in different oxygenation conditions (anoxia, hypoxia, normoxia). Changes in cell oxygenation lead to reactive oxygen species production and oxidative stress. Sigma 1 receptor (Sig1R) is postulated to be stress responding agent and superoxide dismutases (SOD1 and SOD2) are key antioxidant enzymes. It is possible that they participate in tumor cells adaptation to different concentrations of oxygen.
Evaluation of Sig1R, SOD1, and SOD2 expression in different concentrations of oxygen (1%, 10%, 21%) in colon adenocarcinoma cell lines.
SW480 (primary adenocarcinoma) and SW620 (metastatic) cell lines were cultured in standard conditions in Dulbecco's modified Eagle's medium for 5 days, and next cultured in Hypoxic Chamber in 1% O, 10% O, 21% O. Number of living cells was determined by trypan blue assay. Level of mRNA for Sig1R, SOD1, and SOD2 was determined by standard PCR method. Statistical analysis was conducted using Statistica 10.1 software.
We observed significant changes in expression of Sig1R, SOD1, SOD2 due to different oxygen concentrations. ANOVA analysis revealed significant interactions between studied parameters mainly in hypoxia conditions in SW480 cells and between Sig1R and SOD2 in SW620 cells. It also showed that changes in expression of studied proteins depend significantly on type of the cell line.
Changes of Sig1R and SOD2 expression point to mitochondria as main organelle responsible for survival of tumor cells exposed to hypoxia or oxidative stress. Studied proteins are involved in intracellular response to stress related with different concentrations of oxygen.
由于与毛细血管的距离不同，肿瘤细胞存在于不同的氧合条件下（无氧、低氧、正常氧）。细胞氧合的变化会导致活性氧的产生和氧化应激。推测西格玛1受体（Sig1R）是应激反应因子，超氧化物歧化酶（SOD1和SOD2）是关键的抗氧化酶。它们有可能参与肿瘤细胞对不同氧浓度的适应。
评估结肠腺癌细胞系在不同氧浓度（1%、10%、21%）下Sig1R、SOD1和SOD2的表达。
将SW480（原发性腺癌）和SW620（转移性）细胞系在标准条件下于杜尔贝科改良的 Eagle 培养基中培养5天，然后在缺氧培养箱中分别于1% O₂、10% O₂、21% O₂条件下培养。通过台盼蓝试验测定活细胞数量。采用标准PCR方法测定Sig1R、SOD1和SOD2的mRNA水平。使用Statistica 10.1软件进行统计分析。
我们观察到由于氧浓度不同，Sig1R、SOD1、SOD2的表达有显著变化。方差分析显示，主要在SW480细胞的低氧条件下以及SW620细胞中Sig1R和SOD2之间，研究参数之间存在显著相互作用。还表明所研究蛋白质表达的变化显著取决于细胞系类型。
Sig1R和SOD2表达的变化表明线粒体是负责暴露于低氧或氧化应激的肿瘤细胞存活的主要细胞器。所研究的蛋白质参与了细胞内对与不同氧浓度相关应激的反应。
