Fragou Aikaterini, Tzimagiorgis Georgios, Karageorgopoulos Christos, Barbetakis Nikolaos, Lazopoulos Axilleas, Papaioannou Maria, Haitoglou Costas, Kouidou Sofia
Laboratory of Biological Chemistry, Medical Department, School of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece.
Department of Thoracic Surgery, Theagenion Anticancer Hospital, Thessaloniki 54007, Greece.
Mol Med Rep. 2017 Apr;15(4):1455-1460. doi: 10.3892/mmr.2017.6162. Epub 2017 Feb 2.
Modification of p53 expression levels and its principle apoptosis and cell cycle regulatory partners, mouse double minute 2 homolog (MDM‑2) and p21, has been previously reported in various types of cancer. In the current study, the expression of Δ133p53 isoforms was investigated in lung carcinomas with respect to the expression of the aforementioned genes. The expression of p53 full‑length transcript and Δ133p53 isoforms α, β and γ transcripts, MDM‑2 and p21 transcripts were determined by reverse transcription‑quantitative polymerase chain reaction, in total RNA isolated from 17 lung carcinoma specimens and 17 corresponding adjacent non‑cancerous tissues. RNA expression analysis was performed according to the Pfaffl equation and Rest tool using β‑actin as a reference gene. Detection of the above proteins was additionally performed by western blotting. Significant overexpression of the Δ133p53 mRNAs was observed in cancerous as compared with adjacent non‑cancerous tissues (3.94‑fold), whereas full‑length p53 and MDM‑2 expression exhibited a smaller, however significant, increase. The expression of the p21 transcript was significantly reduced in cancerous specimens. Δ133p53 and p21 expression levels varied in parallel, however were not significantly correlated. p53 full‑length protein expression observed by western blot analysis strongly varied from the Δ133p53 isoforms, however MDM‑2 protein isoforms were not detectable and p21 protein was more abundant in non‑cancerous tissues. In conclusion, Δ133p53 mRNA levels is suggested as a potentially useful marker of malignancy in lung cancer. The absence of Δ133p53 protein in lung carcinomas, which overexpress Δ133p53 transcripts, may indicate the role of the latter in post‑transcriptional regulation through RNA interference in the cell cycle and apoptosis.
先前已有报道称,在各类癌症中,p53表达水平及其主要的凋亡和细胞周期调控伙伴——小鼠双微体2同源蛋白(MDM-2)和p21发生了改变。在本研究中,针对上述基因的表达情况,对Δ133p53亚型在肺癌中的表达进行了研究。通过逆转录定量聚合酶链反应,测定了从17例肺癌标本及17例相应的癌旁非癌组织中分离出的总RNA中p53全长转录本、Δ133p53亚型α、β和γ转录本、MDM-2和p21转录本的表达。以β-肌动蛋白作为参照基因,根据Pfaffl方程和Rest工具进行RNA表达分析。另外,通过蛋白质印迹法检测上述蛋白质。与癌旁非癌组织相比,癌组织中Δ133p53 mRNA显著过表达(3.94倍),而全长p53和MDM-2表达虽有较小但显著的增加。癌组织标本中p21转录本的表达显著降低。Δ133p53和p21的表达水平呈平行变化,但无显著相关性。蛋白质印迹分析观察到的p53全长蛋白表达与Δ133p53亚型有很大差异,然而未检测到MDM-2蛋白亚型,且p21蛋白在非癌组织中更为丰富。总之,Δ133p53 mRNA水平被认为是肺癌中一种潜在有用的恶性肿瘤标志物。在过表达Δ133p53转录本的肺癌中不存在Δ133p53蛋白,这可能表明后者在细胞周期和凋亡中通过RNA干扰参与转录后调控。