靶向多梳蛋白1增强子的RNA干扰在肺癌中发挥抗肿瘤作用。
RNA interference targeting enhancer of polycomb1 exerts anti-tumor effects in lung cancer.
作者信息
Che Chunli, Zhang Lijuan, Huo Jianmin, Zhang Yimei
机构信息
Department of Respiratory Medicine, First Clinical Medical College Affiliated to Harbin Medical University Harbin 150001, China.
出版信息
Int J Clin Exp Pathol. 2015 Jan 1;8(1):361-7. eCollection 2015.
BACKGROUND AND AIM
Lung cancer is one of leading malignant tumor worldwide with a high mortality rate. A new therapy target, enhancer of polycomb1 (EPC1) knocked down by short hairpin RNA (shRNA) interference technology, for lung cancer was established to investigate its effects on lung cancer in present study.
METHODS
RNA interference technology was applied to down-regulate the expression of EPC1 by specific-shRNA with lentivirus vector in neoplastic human alveolar basal epithelial cells (A549 cells). The survival rate and apoptosis were respectively measured by MTT and Flow Cytometry to evaluate the effects of shRNA EPC1 on cells. Mice xenografts of HCT116 cells with shRNA EPC1 were also established to assess the effect on tumor growth. The levels of AKT and p65 were detected by western blotting.
RESULTS
The down-regulation of EPC1 by specific-shRNA with lentivirus vector was significantly decreased the survival rate and apoptosis of A549 cells, and the tumors in EPC1 shRNA transfection group had a significant lower size and weight compared with the ones with control shRNA. The protein expression of p-AKT and p65 was reduced by EPC1 shRNA in both in vitro and in vivo experiments.
CONCLUSION
Silencing EPC1 by shRNA technology had the inhibition effects on cell proliferation and tumor growth in lung cancer, which provided a new potential target for treatment of cancers.
背景与目的
肺癌是全球主要的恶性肿瘤之一，死亡率很高。本研究利用短发夹RNA（shRNA）干扰技术构建了一种新的肺癌治疗靶点——多梳蛋白1增强子（EPC1）敲低模型，以研究其对肺癌的影响。
方法
应用RNA干扰技术，通过慢病毒载体携带特异性shRNA下调人肺泡基底上皮癌细胞（A549细胞）中EPC1的表达。采用MTT法和流式细胞术分别检测细胞存活率和凋亡情况，以评估shRNA EPC1对细胞的影响。还构建了携带shRNA EPC1的HCT116细胞小鼠异种移植瘤模型，以评估其对肿瘤生长的影响。通过蛋白质免疫印迹法检测AKT和p65的水平。
结果
慢病毒载体携带特异性shRNA下调EPC1表达后，A549细胞的存活率显著降低，凋亡增加，且EPC1 shRNA转染组的肿瘤体积和重量明显低于对照shRNA组。在体外和体内实验中，EPC1 shRNA均降低了p-AKT和p65的蛋白表达。
结论
shRNA技术沉默EPC1对肺癌细胞增殖和肿瘤生长具有抑制作用，为癌症治疗提供了一个新的潜在靶点。
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