Autophagy induction by low-dose cisplatin: The role of p53 in autophagy
Autophagy induction by low-dose cisplatin: The role of p53 in autophagy
复制标题
DOI：
10.3892/or.2013.2809
复制
发表时间：
2014-01-01
期刊：
影响因子：
4.2
通讯作者：
Yang, Sei-Hoon
中科院分区：
文献类型：
--
作者：
Cho, Kyung-Hwa;Park, Ji-Hye;Yang, Sei-Hoon
大多数肺癌的化疗治疗使用顺铂;然而，它的使用是有限的，因为它有几个副作用。自噬（或II型细胞死亡）是程序性细胞死亡发生的重要机制。本研究的目的是确定低剂量顺铂治疗是否诱导肺癌细胞自噬。我们还研究了自噬抑制是否导致p53介导的细胞凋亡。用5或20 M顺铂处理NCI-H460（野生型p53）和NCI-H1299（无效型p53）细胞12、24或48 h。进行MTT测定以测量顺铂处理后的细胞活力。为了检测顺铂诱导的自噬，检查细胞形态学（自噬空泡）和LC 3定位。使用3-甲基腺嘌呤（3-MA）通过FACS分析检测膜联蛋白V（+）、碘化丙啶（PI）（-）和吖啶橙子（+）细胞来确定自噬抑制的结果。为了确定顺铂是否诱导自噬，我们研究了p53作为细胞存活调节因子在自噬中的作用。低剂量的顺铂（5 M）诱导细胞死亡，这是增强了3-MA在两个细胞系。自噬空泡和胞质LC 3的形成在H460细胞中比在H1299细胞中更明显。与H1299细胞相比，低剂量顺铂在H460中诱导的自噬增加了2倍。然而，细胞凋亡试验显示两种细胞系之间无差异。3-MA预处理后，顺铂诱导的自噬被发现显着减少（3倍减少），野生型p53相比，空型p53细胞。然而，顺铂诱导的凋亡增加野生型p53相比，空型p53细胞。低剂量顺铂诱导肺癌细胞自噬和抑制自噬后的凋亡转移可能是通过激活p53介导的。
The majority of chemotherapy treatments for lung cancer use cisplatin; however, its use is limited as it has several side-effects. Autophagy (or type II cell death) is an important mechanism by which programmed cell death occurs. The purpose of this study was to determine whether low-dose cisplatin treatment induces autophagy in lung cancer cells. We also examined whether autophagy inhibition results in p53-mediated apoptosis. NCI-H460 (wild-type p53) and NCI-H1299 (null-type p53) cells were treated with 5 or 20 M cisplatin for 12, 24 or 48 h. An MTT assay was performed to measure the cell viability following cisplatin treatment. To detect cisplatin-induced autophagy, cell morphology (autophagic vacuole) and LC3 localization were examined. The outcome of autophagy inhibition was determined using 3-methyladenine (3-MA) to detect Annexin V (+), propidium iodide (PI) (-) and acridine orange (+) cells by FACS analysis. To determine whether cisplatin induced autophagy, we examined the role of p53 as a cell survival regulator in autophagy. Low-doses of cisplatin (5 M) induced cell death and this was augmented by 3-MA in both cell lines. Autophagic vacuoles and cytoplasmic LC3 formation was more evident in H460 cells than in H1299 cells. The induction of autophagy by low-dose cisplatin was increased by 2-fold in H460 compared to H1299 cells. However, the tests for apoptosis showed no difference between the 2 cell lines. Following 3-MA pretreatment, cisplatin-induced autophagy was found to be markedly reduced (a 3-fold reduction) in wild-type p53 compared to null-type p53 cells. However, cisplatin-induced apoptosis increased in wild-type p53 compared to null-type p53 cells. Autophagy induction and apoptotic shift after autophagy inhibition may be mediated by p53 activation in lung cancer cells treated with low-dose cisplatin.