论文部分内容阅读
目的探讨利用自主研发的肺癌细胞富集检测试剂盒检测非小细胞肺癌(NSCLc)患者循环肿瘤细胞(CTCs)的可行性及临床价值。方法采用肺癌细胞富集检测试剂盒检测4组标本(A组:Ⅰ期NSCLC组,18例;B组:Ⅱ～Ⅳ期NSCLC组,33例;C组:肺部良性病变组,20例;D组:健康志愿者组,20例)外周血中的肿瘤细胞;同时,采用单纯免疫细胞化学(ICC)法及巢式荧光逆转录聚合酶链反应(RT-PCR)方法检测细胞角蛋白19(CK19 mRNA)、肺特异性X蛋白(LUNX mRNA)的表达作对照。结果采用肺癌细胞富集检测试剂盒检测,A、B、C、D组患者的CK阳性细胞检出率分别为33.3%、60.6%、0%和0%。采用巢式RT-PCR方法检测,A、B、C、D组患者CK19 mRNA表达的阳性率分别为38.9%、63.6%、20.0%和0%,A、B、C、D组患者LUNX mRNA表达的阳性率分别为44.4%、69.7%、10.0%和0%。采用单纯ICC法检测,4组患者的CTCs检出率均为0%。肺癌细胞富集检测试剂盒与单纯ICC法检测比较,差异有统计学意义(P<0.05);肺癌细胞富集检测试剂盒与巢式RT-PCR方法检测比较,差异无统计学意义(P>0.05);A组与B、C、D组患者CK阳性细胞的检出率比较,差异均有统计学意义(P<0.05);阳性率与病理类型、细胞分化程度和临床分期有密切关系(P<0.05)。结论开发的肺癌细胞富集检测试剂盒是检测早期NSCLC患者CTCs的敏感方法,有助于发现隐形微转移、重新确定临床分期及指导患者术后的个体化治疗。
Objective To explore the feasibility and clinical value of detecting circulating tumor cells (CTCs) in patients with non-small cell lung cancer (NSCLc) using the self-developed lung cancer cell enrichment detection kit. Methods Four sets of specimens were collected by lung cancer cell enrichment assay kit (group A: stage Ⅰ NSCLC group, 18 cases; group B: stage Ⅱ ~ Ⅳ NSCLC group, 33 cases; group C: lung benign lesion group, 20 cases; (Group D: healthy volunteers, 20 cases). Cytokeratin 19 was detected by immunocytochemistry (ICC) and nested fluorescent reverse transcription polymerase chain reaction (RT-PCR) (CK19 mRNA), lung X protein (LUNX mRNA) expression as a control. Results The detection rate of CK positive cells in group A, B, C and D was 33.3%, 60.6%, 0% and 0% respectively by using lung cancer cell enrichment detection kit. The positive rates of CK19 mRNA expression in group A, B, C and D were 38.9%, 63.6%, 20.0% and 0% respectively by nested RT-PCR. The expression of LUNX mRNA in group A, B, C and D The positive rates were 44.4%, 69.7%, 10.0% and 0% respectively. Using ICC assay alone, the detection rates of CTCs in all four groups were 0%. The enrichment detection kit of lung cancer cells was significantly higher than that of ICC alone assay (P <0.05). There was no significant difference between the detection kit of lung cancer cells and nested RT-PCR (P> 0.05). The positive rates of CK positive cells in group A, B, C and D were significantly different (P <0.05). The positive rate was closely related to the pathological type, cell differentiation and clinical stage (P < P <0.05). CONCLUSION: The developed lung cancer cell enrichment detection kit is a sensitive method for detecting CTCs in patients with early stage NSCLC. It is helpful to detect invisible micrometastases, re-determine the clinical stage and guide the individualized postoperative treatment.