Enhancement of Apo2L/TRAIL (tumor necrosis factor-related apoptosis-inducing ligand)-induced apoptosis in non-small cell lung cancer cell lines by chemotherapeutic agents without correlation to the expression level of cellular protease caspase-8 inhibitory protein
Enhancement of Apo2L/TRAIL (tumor necrosis factor-related apoptosis-inducing ligand)-induced apoptosis in non-small cell lung cancer cell lines by chemotherapeutic agents without correlation to the expression level of cellular protease caspase-8 inhibitory protein
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DOI：
10.1067/mtc.2002.119694
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发表时间：
2002-01-01
影响因子：
6
通讯作者：
Schmid, RA
中科院分区：
文献类型：
--
作者：
Frese, S;Brunner, T;Schmid, RA
目的：Apo 2L/肿瘤坏死因子相关凋亡诱导配体（tumor necrosis factor-related apoptosis-inducing ligand，TRAIL）是一种特异性促进肿瘤细胞凋亡的潜在抗癌药物。因为不是所有的癌细胞都对Apo 2L/TRAIL敏感，所以我们的研究的目的是确定非小细胞肺癌细胞是否可以被化疗剂敏化Apo 2L/TRAIL诱导的凋亡。此外，测量半胱天冬酶抑制细胞蛋白酶半胱天冬酶-8抑制蛋白（C-FLIP）的内源性表达水平，以研究对Apo 2L/TRAIL的部分抗性。6株人肺癌细胞系（A549、NCI-H358、Calu 1、Calu 6、SkMes 1和SkLu 1）与可溶性Apo 2L/TRAIL和两种不同浓度的顺铂、紫杉醇、多柔比星、5-氟尿嘧啶、和喜树碱。24小时后，通过膜联蛋白V/碘化丙啶染色，随后进行FACScan分析来测量凋亡率。结果：单独用Apo 2L/TRAIL处理肺癌细胞导致四种细胞系中的细胞凋亡（P <0.001）。Apo 2L/7 RAIL与化疗药物联合应用可显著提高细胞凋亡率。统计学分析显示Apo 2L/TRAIL与1.8mmol/L喜树碱和100 mumol/L顺铂的联合作用在6种细胞系中的4种中均具有协同效应（P <0.002）。Western blot分析显示，对Apo 2L/TRAIL的致敏与细胞蛋白酶caspase-8抑制蛋白的表达无关。此外，没有增加细胞蛋白酶caspase-8抑制蛋白水平相对于那些在正常肺组织中可以发现在非小细胞肺癌标本从12 patients.Conclusion：Apo 2L/TRAIL诱导的非小细胞肺癌细胞系的凋亡显着增强化疗药物。对Apo 2L/TRAIL的抗性和敏感性与细胞蛋白酶caspase-8抑制蛋白的内源性表达水平无关，这意味着在非小细胞肺癌中，其他机制负责抑制Apo 2L/TRAIL通路。尽管其分子机制尚不清楚，但Apo 2L/TRAIL联合化疗可能是治疗非小细胞肺癌的一种有前景的治疗方式。
Objective: Apo2L/tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potential anticancer drug that promotes apoptosis specifically in tumor cells. Because not all cancer cells are susceptible to Apo2L/TRAIL, the aim of our study was to determine whether non-small cell lung cancer cells can be sensitized by chemotherapeutic agents for Apo2L/TRAIL-induced apoptosis. In addition, endogenous expression levels of the caspase-inhibiting cellular protease caspase-8 inhibitory protein (C-FLIP) were measured to investigate partial resistance to Apo2L/TRAIL.Methods: Six human lung cancer cell lines (A549, NCI-H358, Calu1, Calu6, SkMes1, and SkLu1) were incubated with soluble Apo2L/TRAIL and two different concentrations each of cisplatin, paclitaxel, doxorubicin, 5-fluorouracil, and camptothecin. After 24 hours the rate of apoptosis was measured by annexin V/propidium iodide staining followed by FACScan analysis. Expression levels of C-FLIP in cell lines and lung cancer biopsy specimens were determined by Western blotting.Results: Treatment of lung cancer cells with Apo2L/TRAIL alone resulted in apoptotic cell death in four cell lines (P < .001). Combining Apo2L/7RAIL and chemotherapeutic agents enhanced the rate of apoptosis significantly. Statistical analysis revealed a synergistic effect of Apo2L/TRAIL in combination with 1.8 mmol/L camptothecin and 100 mumol/L cisplatin, each in four of the six cell lines (P < .002). Western blot analysis showed that sensitization to Apo2L/TRAIL did not correlate with the expression of cellular protease caspase-8 inhibitory protein. Furthermore, no increased cellular protease caspase-8 inhibitory protein levels relative to those in normal lung tissue could be found in non-small cell lung cancer specimens from 12 patients.Conclusion: Apo2L/TRAIL-induced apoptosis in non-small cell lung cancer cell lines is significantly enhanced by chemotherapeutic agents. Resistance and sensitization to Apo2L/TRAIL are not correlated with the endogenous expression level of cellular protease caspase-8 inhibitory protein, implying that in non-small cell lung cancer other mechanisms are responsible for inhibition of the Apo2L/TRAIL pathway. Even though the molecular mechanism remains unclear, the combination of Apo2L/TRAIL with chemotherapy may be a promising treatment modality for non-small cell lung cancer.