Apoptosis as a measure of chemosensitivity to cisplatin and taxol therapy in ovarian cancer cell lines
Apoptosis as a measure of chemosensitivity to cisplatin and taxol therapy in ovarian cancer cell lines
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DOI：
10.1006/gyno.1997.4637
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发表时间：
1997-04-01
影响因子：
4.7
通讯作者：
GercelTaylor, C
中科院分区：
文献类型：
--
作者：
Gibb, RK;Taylor, DD;GercelTaylor, C
客观的。在四种人卵巢癌细胞系中研究了顺铂和紫杉醇诱导的细胞凋亡，以评估细胞凋亡作为化疗敏感性的量度。方法。通过磺胺罗丹明 B 测定测定 OVCAR-3、SKOV-3、UL-1 和 UL-2 细胞对顺铂或紫杉醇的体外敏感性。通过在暴露24小时和48小时后用顺铂和/或紫杉醇处理后的DNA断裂来研究细胞凋亡的诱导。通过二苯胺测定进一步定量 DNA 片段，并通过流式细胞术测定细胞周期 G1、G2/M 和 S 期的细胞比例。通过蛋白质印迹检查p53基因产物的存在。结果。四种细胞系代表对顺铂和紫杉醇的不同敏感性（顺铂的LD50范围，5-30μg/ml；紫杉醇，30-1000nM）。 UL-2代表一种耐药细胞系，与其他细胞系相比，其对紫杉醇的耐药性是其10-30倍，对顺铂的耐药性是其6倍。细胞凋亡的证明与细胞系对 OVCAR-3 和 UL-2 的顺铂和紫杉醇的敏感性相关。当用顺铂或紫杉醇处理 24 或 48 小时时，OVCAR-3 的 DNA 片段均匀存在。 UL-2 在用顺铂或紫杉醇治疗 24 或 48 小时后未显示细胞凋亡。当使用顺铂和紫杉醇进行测序实验时，DNA 片段化与细胞毒性测定相关，但在 UL-1 细胞中，在顺铂和紫杉醇的不同相互作用中没有观察到显着差异。紫杉醇预处理通常会导致细胞毒性以时间表依赖性方式增强，并且片段化增加；顺铂预处理始终导致碎片减少。片段化 DNA 的定量与凝胶电泳上观察到的相关。 OVCAR-3和UL-1在24小时时表现出相对于基线的最大变化（分别是基线的3.8和3.7倍），而UL-2在治疗后相对于基线几乎没有变化。 G1 期停滞在 OVCAR-3 和 SKOV-3 细胞中更容易发生。 UL-2细胞进入G1停滞的细胞比例几乎没有变化，但G2/M比例显着增加。在OVCAR-3、UL-1和UL-2细胞中，我们证明存在异常表达的p53基因产物，而在SKOV-3细胞中未检测到p53。结论。我们的研究结果表明，顺铂和紫杉醇实现显着细胞毒性的能力可能与诱导细胞凋亡直接相关。然而，细胞和遗传特征决定了这些治疗的最终结果。 (C) 1997 年学术出版社。
Objective. Cisplatin- and Taxol-induced apoptosis was studied in four human ovarian cancer cell lines to evaluate apoptosis as a measure of chemosensitivity.Methods. In vitro sensitivities of OVCAR-3, SKOV-3, UL-1, and UL-2 cells to cisplatin or Taxol were determined by the sulforhodamine B assay. Induction of apoptosis was studied by DNA fragmentation following treatment with cisplatin and/or Taxol after 24- and 48-hr exposure. DNA fragmentation was further quantitated by the diphenylamine assay and the proportion of cells in the G1, G2/M, and S phase of the cell cycle was determined by flow cytometry. Presence of the p53 gene product was examined by Western blotting.Results. The four cell lines represent various sensitivities to cisplatin and Taxol (LD50 range for cisplatin, 5-30 mu g/ml; Taxol, 30-1000 nM). UL-2 represents a resistant cell line which was 10-30 times resistant to Taxol and 6 times resistant to cisplatin when compared to the others. Demonstration of apoptosis correlated with the sensitivity of the cell lines to both cisplatin and Taxol for OVCAR-3 and UL-2. DNA fragmentation of OVCAR-3 was uniformly present when treated with cisplatin or Taxol, at 24 or 48 hr. UL-2 demonstrated no apoptosis after 24 or 48 hr of treatment with either cisplatin or Taxol. When sequencing experiments were performed with cisplatin and Taxol, DNA fragmentation correlated with the cytotoxicity assays, except in UL-1 cells where no significant difference was observed in different interactions of cisplatin and Taxol. Pretreatment with Taxol generally resulted in enhanced cytotoxicity in a schedule-dependent manner, and increased fragmentation was demonstrated; cisplatin pretreatment consistently resulted in decreased fragmentation. Quantitation of the fragmented DNA correlated with that seen on gel electrophoresis. OVCAR-3 and UL-1 demonstrated the greatest change from baseline at 24 hr (3.8 and 3.7 times baseline, respectively), whereas UL-2 had little change from the baseline following treatment. G1 arrest occurred more readily in OVCAR-3 and SKOV-3 cells. UL-2 cells had very little change in the proportion of cells entering G1 arrest, but had a significant increase in the G2/M proportion. In OVCAR-3, UL-1, and UL-2 cells, we demonstrated the presence of an aberrantly expressed p53 gene product, while no p53 was detected in the SKOV-3 cells.Conclusions. Our findings indicate that the ability to achieve significant cytotoxicity by cisplatin and Taxol may be directly related to the induction of apoptosis; however, cellular and genetic characteristics determine the eventual outcome of these treatments. (C) 1997 Academic Press.