Perillyl alcohol-mediated inhibition of lung cancer cell line proliferation: potential mechanisms for its chemotherapeutic effects.
Perillyl alcohol-mediated inhibition of lung cancer cell line proliferation: potential mechanisms for its chemotherapeutic effects.
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DOI：
10.1016/j.taap.2003.11.013
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发表时间：
2004-03
影响因子：
3.8
通讯作者：
Mian Xu;Heather S. Floyd;Suzanne M Greth;Wen‐chi Chang;K. Lohman;R. Stoyanova;G. Kucera;T. Kute;M. Willingham;M. S. Miller
中科院分区：
文献类型：
--
作者：
Mian Xu;Heather S. Floyd;Suzanne M Greth;Wen‐chi Chang;K. Lohman;R. Stoyanova;G. Kucera;T. Kute;M. Willingham;M. S. Miller
紫苏醇(POH)目前正在作为抗癌剂进行临床试验，尽管其作用机制尚未确定。我们用POH处理两种人肺癌细胞系H322和H838，以确定其抗肿瘤特性。用硫代罗丹明B(SRB)细胞增殖实验检测0.25、0.5、0.75、1.0和1.5 mM的POH作用1天和5天后的细胞增殖情况。治疗1d后，最低浓度与最高浓度之间的差异不明显。然而，5天后，两种细胞系的细胞增殖均呈剂量依赖性下降，降幅在15%至83%之间。克隆形成实验证实了这些结果--虽然POH在暴露1天后没有明显的影响，但在处理5天后，克隆形成随剂量的增加而减少，从15%到100%不等。时间推移视频显微镜显示1.5 mM POH作用24-48小时内可见明显的细胞凋亡。当POH激活caspase-3活性3-6倍时，多聚ADP-核糖聚合酶(PARP)被裂解，caspase-3活性增加3-6倍。4‘，6-二氨基-2-苯基吲哚(DAPI)染色证实了POH诱导的细胞典型的凋亡特征。在1.5 mM POH作用8和24(H322)或8和48(H838)h后进行DNA芯片表达分析。虽然在POH处理后的不同时间，两种细胞中有大量基因表达上调或下调，但在两种细胞中，只有8种基因的表达水平在两个时间点都是上调或下调的。这些基因作为POH作用的潜在媒介的意义仍不确定，但微阵列表达分析检测到的通常上调或下调的基因数量有限，表明POH可能通过转录后机制介导其作用。我们的结果提示，POH可能作为一种潜在的抗癌药物，刺激或敏化肺肿瘤细胞的凋亡，这种作用可能依赖于肿瘤细胞中存在的遗传损伤。
Perillyl alcohol (POH) is currently being tested in clinical trials as an anticancer agent, though its mechanism of action has not been definitively established. We treated two human lung cancer cell lines, H322 and H838, with POH to determine its antitumor properties. A sulforhodamine B (SRB) cell proliferation assay was used to determine the effects of POH after 1 and 5 days of treatment with 0.25, 0.5, 0.75, 1.0, and 1.5 mM POH. After 1 day of treatment, little difference could be seen between the lowest and highest concentrations of POH. However, after 5 days, both cell lines showed a dose-dependent decrease in cell proliferation that ranged from 15% to 83%. A clonogenic assay confirmed these results—while there was no significant effect of POH after 1 day of exposure, a dose-dependent decrease in colony formation, ranging from 15% to 100%, was seen after 5 days of treatment. Time-lapse video microscopy revealed that apoptotic cells were evident within 24–48 h of treatment with 1.5 mM POH. The appearance of apoptotic cells was preceded by increased caspase-3 activity and cleavage of poly (ADP-ribose) polymerase (PARP) as POH activated caspase-3 activity 3–6-fold. Nuclear staining with 4′,6-diamidino-2-phenylindole (DAPI) confirmed the classical characteristics of apoptosis in POH-treated cells. DNA microarray expression analysis was performed following 8 and 24 (H322) or 8 and 48 (H838) h of treatment with 1.5 mM POH. While a large number of genes were up- or downregulated in the two cell lines at various times after POH treatment, the levels of expression of only eight genes were up- or down-related in both cell lines at both of the time points examined. The significance of these genes as potential mediators of POH action is still uncertain, but the limited number of commonly up- or downregulated genes detected by microarray expression analysis suggests that POH may mediate its effects via posttranscriptional mechanisms. Our results suggest that POH may have potential use as an anticancer drug that stimulates or sensitizes lung tumor cells to apoptosis, and this effect may depend on genetic lesions present in tumor cells.