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目的研究非小细胞肺癌(non-small cell lung cancer,NSCLC)的肿瘤干细胞和非肿瘤干细胞中表皮生长因子受体(epidermal growth factor receptor,EGFR)基因状态。方法收集2010年6月至2011年8月四川省肿瘤医院及四川大学华西医院胸外科NSCLC新鲜手术标本35例,流式细胞术分选出CD133+细胞和CD133-细胞,采用扩增阻滞突变系统(amplification refractory mutation system,ARMS)分别检测EGFR基因突变,观察其有无差异。结果 35例NSCLC样本中均检测到CD133的表达。35例CD133+细胞中EGFR基因突变检出率为28.6%,腺癌突变率高于鳞癌(P<0.05),而在不同性别、年龄、有无吸烟史中无统计学差异(P>0.05);CD133-细胞中EGFR基因突变检出率为42.9%,腺癌、无或少吸烟史者突变率高(P<0.05),而不同性别、年龄的突变率无统计学差异(P>0.05)。11例患者标本的CD133+/-细胞存在EGFR基因异质性,占31.4%(11/35)。24例患者标本CD133+/-细胞的EGFR基因状态相同(18例患者中CD133+和CD133-细胞EGFR基因均为野生型,6例患者中CD133+和CD133-细胞EGFR基因为相同的敏感性突变)。结论NSCLC患者中CD133+细胞和CD133-细胞中存在EGFR基因异质性。
Objective To study the gene status of epidermal growth factor receptor (EGFR) in tumor stem cells and non-tumor stem cells of non-small cell lung cancer (NSCLC). Methods Thirty-five fresh surgical specimens of NSCLC from Sichuan Cancer Hospital and Sichuan Huaxi Hospital were collected from June 2010 to August 2011. CD133 + cells and CD133- cells were sorted out by flow cytometry. (amplification refractory mutation system, ARMS) were detected EGFR gene mutations observed whether the difference. Results The expression of CD133 was detected in all 35 NSCLC samples. The mutation rate of EGFR gene in 35 cases of CD133 + cells was 28.6%, and the mutation rate of adenocarcinoma was higher than that of squamous cell carcinoma (P <0.05). There was no significant difference in gender, age and smoking history (P> 0.05) ; The mutation rate of EGFR gene in CD133- cells was 42.9%. The mutation rate of adenocarcinoma patients with or without smoking history was high (P <0.05), while there was no significant difference in the mutation rates of different sex and age (P> 0.05) . 11 cases of patients with CD133 +/- cells EGFR gene heterogeneity, accounting for 31.4% (11/35). The EGFR gene status of CD133 +/- cells was the same in 24 patients (EGFR gene of both CD133 + and CD133- cells were wild-type in 18 patients and same sensitivity mutation of CD133 + and CD133- cells in 6 patients). Conclusion There is EGFR gene heterogeneity in CD133 + cells and CD133- cells in NSCLC patients.