今日药学

2017, v.27(02) 113-117+126

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人原代肺癌细胞体外长期培养及个体化药敏研究
In Vitro Culture of long-term Patient-derived Conditionally Reprogrammed Cells from Lung Cancer and Application to Individual Chemotherapeutic Susceptibility Studies

黎张燕;唐欲博;罗红鹤;程超;柯尊富;陈孝;
LI Zhangyan;TANG Yubo;LUO Honghe;CHENG Chao;KE Zunfu;CHEN Xiao;The First Affiliated Hospital of Sun Yatsen University;

摘要(Abstract):

目的应用条件性重编程细胞(Conditionally Reprogrammed Cells,CRCs)技术建立高效、稳定的人肺癌细胞体外长期培养体系,并利用该体系进行体外药敏检测,预测患者对常用化疗药物的敏感性,为肺腺癌精准化疗敏感药物的选择提供参考。方法采用胶原酶消化法将肺肿瘤组织分散成单个细胞后,用条件性培养基及3T3饲养细胞诱导培养,观察其生长情况,采用HE染色、免疫组化和免疫荧光法鉴定细胞,MTS药敏试验检测顺铂、奈达铂、卡铂、长春瑞滨4种临床常用化疗药物的敏感性。结果利用CRC技术成功分离培养了14例肺癌细胞,鉴定发现细胞形态、核分裂相以及细胞标记蛋白符合肺癌细胞特征,细胞体外连续培养时间可达5个月以上,并可稳定传代至23代以上。对6例肺腺癌的药敏结果显示,6例原代肺腺癌细胞对顺铂、奈达铂敏感,但对卡铂、长春瑞滨的敏感率较低,这与临床上铂类药物作为肺腺癌一线治疗药、而长春瑞滨作为二线治疗药物相符合。结论 CRCs技术可作为一种体外长期高效培养原代肺癌细胞的方法,应用此方法培养出的原代肿瘤细胞可作为体外药敏检测模型,对肺癌的个体化治疗具有重要意义。
The study aimed to establish stable long-term culture system of human primary lung cancer cells using the conditionally Reprogrammed Cells( CRCs) technology,then apply to anticancer drug susceptibility test which could potentially be used to predict sensitivity to chemotherapy individually. METHODS Lung cancer tissues were collected from patients undergoing surgical resection of lung tumor. Collagenase was used to disperse lung cancer tissue into single cells,and the cells were then plated in conditional medium containing irradiated 3T3 J2 cells. During the culture process,cell morphology was observed under the microscope.HE staining,immunohistochemistry and immunofluorescence were used to identify the cells. Drug susceptibility test was performed with four clinical commonly used chemotherapy drugs( nedaplatin,cis-platinum,carboplatin and vinorelbine) in six cases of primary adenocarcinoma of lung cancer cells. RESULTS Fourteen cases of lung cancer cells were isolated successfully by using CRC technology,the cellular morphology,mitosis and protein expression of cell marker were in line with the characteristics of lung cancer cells. Moreover,the cells were able to be cultured for more than 5 months in vitro,and could be passaged to P23. Drug sensitivity results showed that six cases of primary cells from adenocarcinoma of lung were all sensitive to cis- platinum,nedaplatin,but exhibited low sensitity to carboplatin and vinorelbine,which is consistent with clinical condition. CONCLUSION CRCs technique can be used as an effectively long-term cultivation method for primary lung cancer cells in vitro. Primary tumor cells derived from the established method can be applied to develop models for drug sensitivity test which owns great significance for the individualized treatment.

关键词(KeyWords): 肺癌;肺腺癌;原代细胞;条件重编程细胞;药敏检测;体外培养
lung cancer;adenocarcinoma of lung;primary cells;conditionally reprogrammed cells;drug sensitivity test;in vitro cultures of cells

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作者(Author): 黎张燕;唐欲博;罗红鹤;程超;柯尊富;陈孝;
LI Zhangyan;TANG Yubo;LUO Honghe;CHENG Chao;KE Zunfu;CHEN Xiao;The First Affiliated Hospital of Sun Yatsen University;

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