鼻咽癌肿瘤干细胞分化过程中促血管生成相关因子的表达及其对血管生成细胞的影响

Expression of Angiogenic Association Factors in Differentiating NPC Stem Cells and Promotion of Angiogenesis Cells Colony Formation

目的探讨IL-6作为鼻咽癌复发治疗靶点的可行性。观察鼻咽癌肿瘤干细胞分化初期促血管生成相关因子的表达情况及其对血管生成细胞克隆形成的影响。方法本研究首先采用ELISA检测鼻咽癌肿瘤干细胞分化初期细胞因子IL-6及血管内皮生长因子VEGF的分泌情况;进而采用Real timePCR和Western blot技术对比IL-6干扰前后HIF-1α、STAT3、VEGF的转录和翻译水平,并采用平板克隆形成技术检测不同处理组细胞上清液对人脐静脉内皮细胞(HUVECs)克隆形成能力的影响。结果分化初期的鼻咽癌肿瘤干细胞与未分化细胞及普通鼻咽癌细胞CNE-2细胞株比较,具有更强的分泌IL-6及VEGF的能力,且IL-6、HIF-1α、STAT3、VEGF在转录和翻译水平都有较高表达;利用RNA干扰技术沉默IL-6基因表达后,能够明显降低HIF-1α、STAT3、VEGF的表达;分化初期的鼻咽癌肿瘤干细胞上清液较未分化细胞上清液具有更强的促HUVECs细胞平板克隆形成能力,而在鼻咽癌肿瘤干细胞分化初期沉默IL-6基因后所得到的细胞上清液的促HUVECs细胞克隆形成能力明显降低。结论 IL-6可作为鼻咽癌发生发展的一个治疗靶点,为临床提供治疗参考。在鼻咽癌肿瘤干细胞分化初期促血管生成因子表达升高并且具有较强的促血管生成能力。

Objective To investigate the potential of IL-6 as a therapeutic target for recurrent nasopharyngeal carcinoma. The main purpose of this study is to observe the expression of angiogenic associated factors during nasopharyreal carcinoma （Nl） stem cell early stage differentiation and its effect on an4giogenesis clone cells. Methods The secretion of cytokine IL-6 during nasopharyngeal carcinoma （NPC） stem cell early stage differen- tiation and VEGF was detected by ELISA. The IL-6, HIF-la,STAT 3 and VEGF expression levels were meas- ured by Real time IR and Western blot before and after IL-6 interference The colony forming ability of HU- VECs made by cell supernatant of different groups was detected by plate colony formation assay. Results Com- pared with poor differentiation Nt cell line CNE-2 and undifferentiated NPC cancer stem cells CNE-2S, the early differentiation stage cancer stem cells secreted more IL-6 and VEGF, exhibited enhanced expression of IL-6, HIF-1 a, STAT 3 and VEGF. After silencing IL-6 gene expression through its specific siRNA, those ceils obviously reduced the expression of HIF-la.STAT 3.VEGF genes. Cell culture media of early stage differ- entiation ~ cancer stem cells showed stronger ability to promote HUVEC colony formation than other groups, this promotion ability was sharply decreased with IL-6 gene silencing. Conclusion The findings suggest that IL-6 can be a therapeutic target for nasopharyngeal carcinoma and a reference for clinical treatment. NPC cancer stem cells enhanced angiogenic association factors during early differentiation stage and promoted anglo- genesis in this period.