Department of Oncology, Dezhou People's Hospital, Shandong Province, P.R. China.
Eur Rev Med Pharmacol Sci. 2017 Jun;21(11):2659-2664.
Expression of Notch1 gene in lung cancer A549 cells was reduced using small interfering RNA (small interfering RNA, siRNA) and the effect of Notch1 gene on proliferation and chemo sensitivity of lung cancer A549 cells was studied.
The Notch1 siRNA was transfected into A549 cells by liposome to inhibit the expressions of Notch1 gene in A549 cells. Real-time polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of Notch1 gene and protein. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was used to detect the proliferation of A549 cells. After transfection of Notch1-siRNA, cisplatin was added to each group at a concentration of 4 μg/ml, and cultured for 48 h. MTT assay and 4',6-diamidino-2-phenylindole (DAPI) staining was used to evaluate the change of apoptosis and sensitivity to chemotherapy.
Notch1 gene expression of A594 cells, detected by RT-PCR and Western blot was significantly reduced in transfected cells when compared with the control group (p<0.05). Inhibition of A594 cell proliferation was significantly decreased as detected by MTT (p<0.05), and the MTT assay and DAPI staining showed that Notch1 gene silencing can significantly improve the sensitivity of A549 cells to chemotherapeutic drugs.
The Notch1 siRNA can effectively inhibit the expression of Notch1 gene, inhibit the proliferation of lung cancer A549 cells and increase the sensitivity to chemotherapeutic drugs.
采用小干扰 RNA（small interfering RNA，siRNA）降低肺癌 A549 细胞中 Notch1 基因的表达，研究 Notch1 基因对肺癌 A549 细胞增殖和化疗敏感性的影响。
脂质体转染 Notch1 siRNA 抑制 A549 细胞中 Notch1 基因的表达，采用实时聚合酶链反应（real-time polymerase chain reaction，RT-PCR）和 Western blot 检测 Notch1 基因和蛋白的表达，3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide，MTT）法检测 A549 细胞的增殖情况，转染 Notch1-siRNA 后，各组加入浓度为 4 μg/ml 的顺铂，培养 48 h，采用 MTT 法和 4',6-二脒基-2-苯基吲哚（4',6-diamidino-2-phenylindole，DAPI）染色法评价细胞凋亡及化疗敏感性的变化。
与对照组比较，转染组 A594 细胞 RT-PCR 和 Western blot 检测 Notch1 基因表达明显降低（p<0.05），MTT 法检测 A594 细胞增殖抑制率明显降低（p<0.05），MTT 法和 DAPI 染色显示 Notch1 基因沉默可显著提高 A549 细胞对化疗药物的敏感性。
Notch1 siRNA 可有效抑制 Notch1 基因的表达，抑制肺癌 A549 细胞的增殖，增加其对化疗药物的敏感性。
