摘要
目的检测吉非替尼敏感性不同的非小细胞肺癌中miR-7的表达差异,并探讨其临床意义。方法采用吉非替尼(Gefitinib)药物大剂量冲击法诱导H827,建立耐吉非替尼肺癌细胞亚系H827-7/GR,有限稀释法将H827-7/GR细胞单克隆化,CCK-8法检测耐药前后细胞株及各单克隆细胞对吉非替尼的敏感性;RT-PCR方法检测H827、H827-7/GR和耐药单克隆细胞株以及其他对吉非替尼敏感性不同的肺癌细胞株A549、H358、H1299、H1650和H1975中miR-7的表达差异。结果 H827/GR耐药指数大于100;获得的6株耐药单克隆细胞株对吉非替尼的半生长抑制浓度(the half growth inhibition concentration,IC50)值不同;和吉非替尼敏感株H827相比,诱导的耐药细胞株H827/GR、耐药单克隆细胞株和吉非替尼耐药株A549、H358、H1299、H1650和H1975中miR-7的相对表达水平均降低(P<0.05)。结论在非小细胞肺癌中,耐药细胞中miR-7的相对表达水平均较敏感细胞系降低,提示miR-7的低表达可能与非小细胞肺癌耐药性相关,其可能是潜在的肺癌药物敏感性预测分子标志物。
Objective To detect the difference of miRNA-7 expression in the non-small cell lung cancer with different sensitivity to gefitinib and explore their clinical significance. Methods The gefitinib resistant lung cancer cell line H827-7/GR was established from H827 by a method of repeated treatment with high dose of gefifinib for a short period. The H827-7/GR cells were monocloned through the method of limiting dilution. CCK- 8 was used to test the sensitivity of H827, H827-7/GR and the monoclonal cells to gefitinib. RT-PCR was used to detect the difference of miRNA-7 expression in H827, H827-7/GR and the monoclonal ceils, and other lung cancer cell lines with different sensitivity to gefitinib, such as A549, H358, H1299, H1650 and H1975. Results The drug resistance index of H827/GR was larger than 100. The half growth inhibition concentrations （ IC50 ） of the six resistant monoclonal cell lines were different. The miRNA-7 expression of H827/GR, resistant monoelonal cell lines, and gefitinib resistant cell lines such as A549, H358, H1299, H1650 and H1975 were all lower than that of the gefitinib sensitive cell line H827 （ P 〈 0.05 ）. Conclusions The miRNA-7 expression of gefitinib resistant cell lines is lower than that of gefitinib sensitive cell lines in the non small cell lung cancer. This suggests that the drug resistance of the non small cell lung cancer may be associated with the low expression of miRNA-7 and it may be a potential molecular marker of the sensitivity of lung cancer to the drugs.
出处
《中国肿瘤外科杂志》
CAS
2015年第2期74-78,共5页
Chinese Journal of Surgical Oncology
基金
国家自然科学基金(81372396)
江苏省自然科学基金(BK20141016)
