Lang Dagmar S, Droemann Daniel, Schultz Holger, Branscheid Detlev, Martin Christian, Ressmeyer Anne R, Zabel Peter, Vollmer Ekkehard, Goldmann Torsten
Clinical and Experimental Pathology, Research Center Borstel, Borstel, Germany.
Respir Res. 2007 Jun 14;8(1):43. doi: 10.1186/1465-9921-8-43.
Non-small cell lung cancer (NSCLC) causes most of cancer related deaths in humans and is characterized by poor prognosis regarding efficiency of chemotherapeutical treatment and long-term survival of the patients. The purpose of the present study was the development of a human ex vivo tissue culture model and the analysis of the effects of conventional chemotherapy, which then can serve as a tool to test new chemotherapeutical regimens in NSCLC.
In a short-term tissue culture model designated STST (Short-Term Stimulation of Tissues) in combination with the novel *HOPE-fixation and paraffin embedding method we examined the responsiveness of 41 human NSCLC tissue specimens to the individual cytotoxic drugs carboplatin, vinorelbine or gemcitabine. Viability was analyzed by LIFE/DEAD assay, TUNEL-staining and colorimetric MTT assay. Expression of Ki-67 protein and of BrdU (bromodeoxyuridine) uptake as markers for proliferation and of cleaved (activated) effector caspase-3 as indicator of late phase apoptosis were assessed by immunohistochemistry. Transcription of caspase-3 was analyzed by RT-PCR. Flow cytometry was utilized to determine caspase-3 in human cancer cell lines.
Viability, proliferation and apoptosis of the tissues were moderately affected by cultivation. In human breast cancer, small-cell lung cancer (SCLC) and human cell lines (CPC-N, HEK) proliferative capacity was clearly reduced by all 3 chemotherapeutic agents in a very similar manner. Cleavage of caspase-3 was induced in the chemo-sensitive types of cancer (breast cancer, SCLC). Drug-induced effects in human NSCLC tissues were less evident than in the chemo-sensitive tumors with more pronounced effects in adenocarcinomas as compared to squamous cell carcinomas.
Although there was high heterogeneity among the individual tumor tissue responses as expected, we clearly demonstrate specific multiple drug-induced effects simultaneously. Thus, STST provides a useful human model to study numerous aspects of mechanisms underlying tumor responsiveness towards improved anticancer treatment. The results presented here shall serve as a base for multiple functional tests of novel chemotherapeutic approaches to NSCLC in the future.
非小细胞肺癌（NSCLC）导致了人类大多数与癌症相关的死亡，其特点是化疗治疗效果和患者长期生存率的预后较差。本研究的目的是开发一种人类离体组织培养模型，并分析传统化疗的效果，从而可作为测试NSCLC新化疗方案的工具。
在一种名为STST（组织短期刺激）的短期组织培养模型中，结合新型的HOPE固定和石蜡包埋方法，我们检测了41个人类NSCLC组织标本对个体细胞毒性药物卡铂、长春瑞滨或吉西他滨的反应性。通过活/死检测、TUNEL染色和比色MTT检测分析细胞活力。通过免疫组织化学评估Ki-67蛋白的表达、BrdU（溴脱氧尿苷）摄取作为增殖标志物以及裂解（活化）效应半胱天冬酶-3作为晚期凋亡指标的表达。通过RT-PCR分析半胱天冬酶-3的转录。利用流式细胞术测定人类癌细胞系中的半胱天冬酶-3。
组织的活力、增殖和凋亡受到培养的适度影响。在人类乳腺癌、小细胞肺癌（SCLC）和人类细胞系（CPC-N、HEK）中，所有3种化疗药物均以非常相似的方式明显降低了增殖能力。在化疗敏感型癌症（乳腺癌、SCLC）中诱导了半胱天冬酶-3的裂解。与鳞状细胞癌相比，药物诱导的人类NSCLC组织中的效应在腺癌中更明显，比化疗敏感肿瘤中的效应不那么明显。
尽管如预期的那样，个体肿瘤组织反应之间存在高度异质性，但我们清楚地同时证明了特定的多种药物诱导效应。因此，STST提供了一个有用的人类模型，用于研究肿瘤对抗癌治疗反应机制的许多方面。此处呈现的结果应作为未来对NSCLC新型化疗方法进行多种功能测试的基础。
