Restoration of wild-type p53 activity in p53-null HL-60 cells confers multidrug sensitivity.
Restoration of wild-type p53 activity in p53-null HL-60 cells confers multidrug sensitivity.
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发表时间：
1998-05
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通讯作者：
J. Ju;D. Banerjee;H. Lenz;K. Danenberg;Thomas D. Schmittgen;C. Spears;A. Schönthal;D. J. Manno;D. Hochhauser;J. Bertino;P. Danenberg
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作者：
J. Ju;D. Banerjee;H. Lenz;K. Danenberg;Thomas D. Schmittgen;C. Spears;A. Schönthal;D. J. Manno;D. Hochhauser;J. Bertino;P. Danenberg
稳定表达转染野生型（wt）p53的HL-60细胞被用于确定wt p53的恢复是否增加了通常缺乏p53活性的细胞的化学敏感性。野生型p53 HL-60转染子（SN 3细胞）比亲本（S）细胞对代表各种作用机制的许多常见抗癌药物更敏感，而用密码子248和143处突变的p53基因转染的HL-60细胞不敏感。由于转染的野生型p53的敏化比率从顺铂的约2倍到胸苷的超过50倍不等。用胸苷酸合成酶抑制剂5-氟-2 '-脱氧尿苷（FdUrd）处理的细胞用于研究各种p53相关基因表达的变化。在每个FdUrd浓度下，观察到SN 3细胞中的凋亡细胞百分比高于S细胞中的凋亡细胞百分比。S细胞具有不可检测的bax水平和高水平的bcl-2，而SN 3细胞具有不可检测的bcl-2水平和可感知的bax基础水平。经FdUrd处理后，SN 3细胞p53和bax水平均升高，但bax的诱导作用快于p53，并抑制了凋亡DNA梯状条带的出现。FdUrd处理诱导p21表达，并增加SN 3细胞的G1期分数，但不诱导p21或改变S细胞中的时相分布。FdUrd处理也诱导了SN 3细胞中cyclin D1的表达和磷酸化，但在S细胞中没有。这些结果表明，转染的野生型p53赋予HL-60细胞的多药敏感性，通过重新调整的凋亡基因的表达，并显示内源性起源的野生型p53的其他特性。
HL-60 cells that stably express transfected wild-type (wt) p53 were used to determine whether restoration of wt p53 increased the chemosensitivity of cells that normally lack p53 activity. The wt p53 HL-60 transfectants (SN3 cells) were more sensitive than the parental (S) cells to a number of common anticancer drugs representing various mechanisms of action, whereas HL-60 cells transfected with p53 genes mutated at codons 248 and 143 were not sensitized. The sensitization ratio due to the transfected wt p53 varied from about 2-fold for cisplatin to over 50-fold for thymidine. Cells treated with the thymidylate synthase inhibitor 5-fluoro-2'-deoxyuridine (FdUrd) were used to study changes in various p53-associated gene expressions. A higher percentage of apoptotic cells among the SN3 cells was observed than among the S cells at each concentration of FdUrd. The S cells had undetectable levels of bax and high levels of bcl-2, whereas the SN3 cells had undetectable levels of bcl-2 levels and appreciable basal levels of bax. After FdUrd treatment of SN3 cells, both p53 and bax levels increased, but the induction of bax was faster than that of p53 and paralleled the appearance of apoptotic DNA laddering. FdUrd treatment induced p21 expression and increased the G1 fraction of the SN3 cells but did not induce p21 or change the phase distribution in the S cells. FdUrd treatment also induced the expression and phosphorylation of cyclin D1 in the SN3 cells but not in the S cells. These results show that transfected wt p53 confers multidrug sensitivity to HL-60 cells by re-adjustment of the expressions of apoptosis genes and displays other properties characteristic of endogenously originated wt p53.