Treatment of radioresistant stem-like esophageal cancer cells by an apoptotic gene-armed,telomerase-specific oncolytic adenovirus
Treatment of radioresistant stem-like esophageal cancer cells by an apoptotic gene-armed,telomerase-specific oncolytic adenovirus
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用带有凋亡基因的端粒酶特异性溶瘤腺病毒治疗放射抗性干细胞样食管癌细胞
DOI：
10.1158/1078-0432.ccr-07-1528
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发表时间：
2008-05-01
影响因子：
11.5
通讯作者：
Chang, Joe Y.
中科院分区：
文献类型：
--
作者：
Zhang, Xiaochun;Komaki, Ritsuko;Chang, Joe Y.
目的：肿瘤干细胞（CSC）可能引起放射抵抗。由于肿瘤干细胞的增殖需要端粒酶的参与，因此携带凋亡肿瘤坏死因子相关凋亡诱导配体和E1 A基因的端粒酶特异性溶瘤腺病毒载体（Ad/TRAIL-E1）可以优先靶向肿瘤干细胞。实验设计：通过分次照射建立了两对亲本和放射抗性（R）食管癌细胞系（Seg-1、Seg-1 R和TE-2、TE-2 R）。通过Western印迹和流式细胞术测量干细胞标志物。使用连续分选来富集干样侧群细胞。在体外和/或体内测定端粒酶活性、转基因表达、抗肿瘤活性、凋亡诱导和病毒复制。Seg-1 R细胞中干细胞标志物β-连环蛋白、Oct 3/4和整合素的表达分别为29.4%、27.5%和97.3%，而Seg-1 R细胞中干细胞标志物β-连环蛋白、Oct 3/4和整合素的表达分别为4.8%、14.9%和97.3%。而Seg-1细胞为45.3%（P < 0.05）。Seg-1 R和TE-2 R细胞的SP水平分别为14.6%和2.7%，而Seg-1和TE-2细胞的SP水平分别为3.4%和0.3%。Seg-1 R SP细胞的系列分选显示SP细胞富集。Seg-1 R、Seg-1 R SP和TE-2 R细胞的端粒酶活性分别显著高于Seg-1、Seg-1 R non-SP和TE-2细胞（P < 0.05）。Seg-1 R和TE-2 R细胞对Ad/TRAIL-E1的敏感性高于亲本细胞。在放射抵抗细胞中发现柯萨奇-腺病毒受体增加和转基因表达升高。Ad/TRAIL-E1可显著抑制Seg-1 R荷瘤小鼠的肿瘤生长，延长其存活时间（P < 0.05），且无明显毒性作用。Ad/TRAIL-E1优先靶向放射抗性CSC样细胞。
Purpose: Radioresistance may be caused by cancer stem cells (CSC). Because CSCs require telomerase to proliferate, a telomerase-specific oncolytic adenoviral vector carrying apoptotic tumor necrosis factor -related apoptosis-inducing ligand and E1A gene (Ad/TRAIL-E1) may preferentially target CSCs.Experimental Design: We established two pairs of parental and radioresistant (R) esophageal carcinoma cell lines (Seg-1, Seg-1R and TE-2,TE-2R) by fractionated irradiation. Stem cell markers were measured by Western blotting and flow cytometry. Serial sorting was used to enrich stem-like side population cells. Telomerase activity, transgene expression, antitumor activity, apoptosis induction, and viral replication were determined in vitro and/or in vivo.Results: Expression of the stem cell markers beta-catenin, Oct3/4, and, integrin in Seg-1R cells was 29.4%, 27.5%, and 97.3%, respectively, compared with 4.8%,14.9%, and 45.3% in Seg-1 cells (P < 0.05). SP levels in Seg-1R and TE-2R cells were 14.6% and 2.7%, respectively, compared with 3.4% and 0.3% in Seg-1 and TE-2 cells. Serial sorting of Seg-1R SP cells showed enrichment of the SP cells. Telomerase activities in Seg-1R, Seg-1R SP, and TE-2R cells were significantly higher than in Seg-1, Seg-1R non-SP, and TE-2 cells, respectively (P < 0.05). Seg-1R and TE-2R cells were more sensitive to Ad/TRAIL-E1 than parental cells. Increased Coxsackie-adenovirus receptor and elevated transgene expressions were found in the radioresistant cells. Ad/TRAIL-E1 resulted in significant tumor growth suppression and longer survival in Seg-1R -bearing mice (P < 0.05) with no significant toxicity.Conclusion: Radioresistant cells established by fractionated irradiation display CSC-like cell properties. Ad/TRAIL-E1 preferentially targets radioresistant CSC-like cells.