Various forms of mutant p53 confer sensitivity to cisplatin and doxorubicin in bladder cancer cells
Various forms of mutant p53 confer sensitivity to cisplatin and doxorubicin in bladder cancer cells
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DOI：
10.1016/s0022-5347(05)66150-2
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发表时间：
2001-07-01
影响因子：
6.6
通讯作者：
Lai, MD
中科院分区：
文献类型：
--
作者：
Chang, FL;Lai, MD
目的：据报道，p53 突变的性质会影响细胞对化疗的反应。我们表征了p53突变对膀胱癌细胞耐药性的影响。材料和方法：将各种人p53突变体（V143A、V173L、H179Q、N247I和R273H）引入TCC-SUP膀胱癌细胞系中以建立稳定的转染子。通过逆转录酶-聚合酶链式反应和免疫细胞化学分析证明了突变体p53的表达。通过台盼蓝排除法测定这些转染子对顺铂和阿霉素的敏感性。通过末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记分析、Hoechst 33258染色和膜联蛋白-V结合测定来表征顺铂和阿霉素介导的细胞死亡。 结果：除p53His273之外的所有形式的突变p53蛋白的表达增强了对顺铂和阿霉素的敏感性。 p53His273 转染子的化学敏感性与亲本TCC-SUP 和对照转染子相似。顺铂诱导的细胞死亡经历细胞凋亡，这分别通过 Hoechst 染色、膜联蛋白-V 测定和末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记来证明。相反，阿霉素诱导的细胞死亡可能通过非凋亡途径发生。结论：这些结果表明p53突变的性质可能影响细胞对抗癌药物的反应，并且许多形式的突变p53蛋白可能通过凋亡或非凋亡途径增强膀胱癌细胞的化疗敏感性。
Purposes: The nature of p53 mutation has been reported to affect cellular responses to chemotherapy. We characterized the impact of p53 mutations on drug resistance in bladder cancer cells.Materials and Methods: Various human p53 mutants (V143A, V173L, H179Q, N247I and R273H) were introduced to the TCC-SUP bladder carcinoma cell line to establish stable transfectants. The expression of mutant p53 was demonstrated by reverse transcriptase-polymerase chain reaction and immunocytochemical analysis. The sensitivity to cisplatin and doxorubicin in these transfectants was determined by trypan blue exclusion. Cell death mediated by cisplatin and doxorubicin was characterized by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling analysis, Hoechst 33258 staining and annexin-V binding assay.Results: The expression of all forms of mutant p53 protein except p53His273 enhanced sensitivity to cisplatin and doxorubicin. The chemosensitivity of p53His273 transfectants is similar to that of parental TCC-SUP and control transfectants. Cisplatin induced cell death undergoes apoptosis, as demonstrated by Hoechst staining, annexin-V assay and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling, respectively. In contrast, doxorubicin induced cell death probably occurs through a nonapoptotic pathway.Conclusions: These results indicated that the nature of p53 mutations may affect the cellular response to anticancer drugs and many forms of mutant p53 protein may enhance chemosensitivity through apoptotic or nonapoptotic pathways in bladder cancer cells.