摘要
目的通过非小细胞肺癌(NSCLC)患者恶性胸腔积液肿瘤细胞的体外原代培养,检测其对常规化疗药物的敏感性,为临床筛选个体化治疗方案提供参考。方法收集20例经病理组织学确诊为NSCLC患者的恶性胸腔积液,通过密度梯度离心法分选后体外短暂培养以去除非肿瘤细胞成分,建立原代细胞系;使用MTT法检测肺癌常规化疗药物作用于肿瘤细胞的半数抑制浓度(IC50),与理论计算获得的血药浓度值(IC50 m)进行比较获得IC50绝对预测值(R),并将其与患者的临床实际疗效进行比对分析。结果 20例恶性胸腔积液样本经预处理及培养4代后均能获得在体外稳定传代的原代肿瘤细胞系,巴氏染色证实其具有癌细胞特征,且显微镜下观察癌细胞纯度近乎100%。MTT法检测结果显示,使用R值时,能将超出测试浓度上限的无效IC50值进一步纳入疗效评估的有效范围;与其中具有完整化疗史的6例患者的实际疗效比对结果显示,当用R值在0.5～2.0和大于2.0分别预测实际疗效为疾病稳定和疾病进展时,其疗效预测的总体一致性为77%(10/13)。结论癌性胸水原代肿瘤细胞培养及其药敏检测在预测NSCLC患者的实际化疗疗效中具有参考价值。
Objective To provide reference for clinical screening of individualized therapy by detecting the drug susceptibility of primary tumor cells derived from malignant pleural effusion(MPE)of the patients with non-small cell lung cancer(NSCLC).Methods MPE specimens were collected from 20 patients diagnosed as NSCLC by histopathology.They were separated by density gradient centrifugation cultured in vitro to remove non-tumor cells,and then primary cell lines were established.The half-inhibitory concentrations(IC50)of conventional anti-tumor chemotherapy drugs on primary tumor cells were determined by the MTT method,and an absolute predictive value(R)was obtained by comparing the IC50 with the theoretically calculated value of maximum plasma concentration(IC50 m).Last,the R value was compared with the actual clinical efficacy of the NSCLC patients.Results After 20 MPE samples were pretreated and cultured for 4 generations,the primary tumor cell lines passaged stably in vitro were established successfully.Papanicolaou staining confirmed that these tumor cell lines had the characteristics of cancer cells,and their purity was nearly 100%under the microscope.The MTT results showed that the invalid IC50 values beyond the upper limit of test concentration could be further included in the following evaluation on the therapeutic effect of patients when R values were used.When R values between 0.5 and 2.0 and more than 2.0 were used for predicting the actual therapeutic effect as disease stability and disease progression,respectively,the overall consistency between R value and actual therapeutic effect was 77%(10/13)in six patients with complete history of chemotherapy.Conclusion The primary culture of tumor cells in MPF and the detection of drug sensitivity have the clinical application value in predicting the actual therapeutic effect of NSCLC patients.
作者
胡嘉华
王海涛
刘志安
赵建华
HU Jiahua;WANG Haitao;LIU Zhian;ZHAO Jianhua(Center of Clinical Laboratory Science,Affiliated Cancer Hospital of Nanjing Medical University/Jiangsu Cancer Hospital/Jiangsu Institute of Cancer Research,Nanjing210009,Jiangsu;CardiothoracicSurgery,Zhejiang Provincial People's Hospital/People's Hospital of Hangzhou Medical College,Hangzhou310014,Zhejiang,China)
出处
《临床检验杂志》
CAS
2019年第2期133-136,共4页
Chinese Journal of Clinical Laboratory Science
基金
江苏省重点研发计划(BE2015718)
浙江省中医药科学研究基金(2019ZA002)
