非小细胞肺癌乏氧相关辐射耐受分子标志物的筛选及深入研究

项目名称： 非小细胞肺癌乏氧相关辐射耐受分子标志物的筛选及深入研究

项目编号： No.30801066

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 谢丽

作者单位： 山东省医学科学院

项目金额： 20万元

中文摘要： 通过对非小细胞肺癌细胞株进行辐射诱导，建立非小细胞肺癌辐射耐受模型，采用体内外放射敏感性检测方法发现该细胞模型与父代细胞相比辐射耐受性显著提高。同时发现辐射耐受细胞的增殖速度减慢，细胞的迁移运动能力、粘附能力和侵袭潜能均增强，辐射耐受细胞SNAI2、Twist、OCT4和Notch1等蛋白表达上调，且干细胞样细胞的比例增加，辐射耐受细胞对化疗药物敏感性降低。利用Illumina人全基因组表达谱芯片技术比较两者之间及其父代细胞辐射前后的基因表达谱，并进行荧光定量RT-PCR验证，发现一批表达差异分子。其中，溶质转运蛋白家族基因在不同的辐射耐受细胞株中均有明显表达差异。选取在三个细胞株中均有差异表达，且在A549/R细胞差异表达基因中倍数最高的基因AKR1C3进行进一步的功能再验证，发现干扰AKR1C3能够增加辐射诱导的细胞凋亡，提高细胞的辐射敏感性，改变细胞周期，这些作用在IL-6诱导的辐射耐受中更加明显。以上结果证实AKR1C3有可能成为放射增敏治疗的靶分子。以上研究已发表SCI论文2篇，还将发表SCI论文2~3篇，参加ASTRO会议交流2篇。相关内容获山东省医学科学院科技成果一等奖。

中文关键词： 非小细胞肺癌; 辐射敏感性; 表达谱芯片; AKR1C3

英文摘要： The radioresistant non-small cell lung cancer (NSCLC) subclones were established by eight rounds of ionizing radiation. Stable and significant radioresistance was observed in the screened subclones using clonogenic assay and apoptosis analysis. The proliferation of radioresistant subclones as to control cells was suppressed. In vitro analysis showed that fractionated irradiation enhanced cell invasion and migration. SNAI2, Twist, OCT4 and Notch1 related to epithelial-mesenchymal transition (EMT) were up-regulated in radioresistant subclones.The proportion of stem cell-like cells in radioresistant subclones also increased. Fractionated irradiation induced radioresistant cancer cell changed their sensitive to chemotherapeutic drugs. Our study showed that radioresistant cancer cells had some CSC characteristics, and fractionated irradiation enhanced invasion and migration of non-small cell lung cancer by inducing EMT. Differential expressed genes were screened by Illumina Human-6 v2 Expression BeadChip consisting of 48,000 transcript probes. Members the of human solute carrier (SLC) gene superfamily had significantly differential expression. AKR1C3, a member of a family of 3 alpha-hydroxysteroid dehydrogenase, was significantly up-regulated in the radioresistant cells. A decreased expression of AKR1C3 by a specific AKR1C3-small interfering RNA (siRNA) significantly enhanced cell radiosensitivity. Furthermore, IL-6-mediated radioresistance was also reduced by knockdown of AKR1C3 using siRNA. The AKR1C3-mediated radioresistance was correlated with a G2/M arrest, a decreased induction of apoptosis. Our results showed that AKR1C3 might be a potential target in the radioresistance of NSCLC, especially in IL-6-mediated radioresistance, via affecting apoptosis and cell cycle. Our studies have been published in two SCI papers. And we will publish two to three more SCI papers in the future. Two papers were selected into ASTRO Conference Poster. Our result won the first prize of scientific and technological achievement by the Shandong Provincial Academy of Medical Sciences.

英文关键词： NSCLC; Radiosensitivity; Microarray; AKR1C3