摘要
目的探讨不同表皮生长因子受体(epidermal growth factor receptor,EGFR)突变型非小细胞肺癌(non small cell lung cancer,NSCLC)细胞EGFR表达水平与放射线敏感性的关系。方法选用6株不同EGFR基因突变的NSCLC细胞:PC-9和HCC827为19外显子突变; H3255为21外显子突变; H1975为21外显子突变伴T790M突变的原发EGFR-TKI耐药细胞; PC-9/AB和PC-9/ZD为19外显子突变伴T790M突变的获得性耐药细胞。NGS测定EGFR基因状况、Western blot法检测EGFR表达量、RNAi沉默EGFR表达、X射线诱导放射抵抗、克隆成型实验检测各细胞放射线敏感性。结果 H1975细胞放射线敏感性显著高于其他5株细胞,HCC827、H3255细胞对放射线中度敏感,PC-9、PC-9/AB、PC-9/ZD细胞对放射线敏感性最低。H1975细胞的EGFR表达量最低,显著低于其他5株细胞,HCC827、H3255、PC-9/ZD细胞EGFR呈中度表达,PC-9、PC-9/AB细胞EGFR表达量最高。RNAi下调PC-9/ZD细胞的EGFR表达后,其放射线敏感性显著提高(P <0. 05)。X线诱导产生放射线抵抗的PC-9/ABIR、PC-9/ZDIR细胞EGFR表达量较亲本的PC-9/AB、PC-9/ZD细胞显著增加(P <0. 05)。结论不同EGFR突变型NSCLC细胞间放射线敏感性存在差异,放射线敏感性与EGFR表达量相关,EGFR表达高对放射线相对抵抗,EGFR表达低则对放射线相对敏感。EGFR表达量有望成为EGFR突变型NSCLC放射线敏感性预测指标。
Objective To investigate the correlation between EGFR gene expression and the radiosensitivity inEGFR-mutant non-small cell lung cancer (NSCLC) cell lines.Methods Human NSCLC cell lines PC-9 andHCC827 with mutations in exon 19 of EGFR; H3255 with mutations in exon 21 of EGFR; H1975 harboring both theL858R mutation in exon 21; T790 M mutation in exon 20 of EGFR; PC-9 /AB and PC-9 /ZD cell lines harboring boththe mutations in exon 19; and the T790M with mutation in exon 20 of EGFR induced by acquired gefitinib-resistancewere used in this study.EGFR gene status was detected by next generation sequencing analysis.Western blot was appliedto verify the expression of EGFR.RNA interference was used to silence EGFR gene expression.X-ray induced radiore-sistance. Radiosensitivity was determined by clone-forming test.Results Among the 6 NSCLC cell lines,H1975 cellwas the most radiosensitive cell line; HCC827 and H3255 cells were moderately sensitive to radiation; and PC-9、PC-9/AB and PC-9 /ZD cells were the least radiosensitive cell lines.The expression of EGFR in H1975 cell was remarkablylower than that in the other five cell lines.HCC827,H3255 and PC-9 /ZD cells expressed the intermediate level of EGFR.PC-9 and PC-9/AB cells expressed the highest level of EGFR.The EGFR expression in PC-9/ZD cell was significantly reduced with RNA interference,accompanying the enhancing of its radiosensitivity.The X-ray induced radioresistance in PC-9/ AB and PC-9/ZD cell lines and reduced the EGFR expression at the same time.Conclusion Theradiosensitivity of NSCLC cells with EGFR gene mutation is closely associated with EGFR expression. The higher expression of EGFR,the lower sensitivity it is to radiation. EGFR expression may be one of the prospective factors for radiosensitivity.
作者
程燕君
谢波
周娟
张为民
CHENG Yan-jun;XIE Bo;ZHOU Juan;ZHANG Wei -min(Guangdong Pharmaceutical University,Guangzhou 510006,Guangdong,China)
出处
《广东医学》
CAS
2018年第24期3595-3599,共5页
Guangdong Medical Journal
基金
国家自然科学基金面上项目(编号:81472172)
关键词
EGFR表达
突变
放射线敏感性
NSCLC
EGFR expression
mutation
radiosensitivity
non-small cell lung cancer
