Expression of the platelet-activating factor receptor enhances benzyl isothiocyanate-induced apoptosis in murine and human melanoma cells.
Expression of the platelet-activating factor receptor enhances benzyl isothiocyanate-induced apoptosis in murine and human melanoma cells.
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DOI：
10.3892/mmr.2015.3371
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发表时间：
2015-07
影响因子：
3.4
通讯作者：
Sahu RP
中科院分区：
文献类型：
--
作者：
Sahu RP
黑色素瘤细胞通常表达血小板活化因子受体（PAF-R），这已被证明会增加转移行为。然而，PAF-R对黑色素瘤对天然细胞毒性药物的反应性的影响仍有待阐明。本研究旨在确定异硫氰酸苄酯（BITC），十字花科蔬菜的一个组成部分，在黑色素瘤细胞表达PAF-R的相对细胞毒性和机制。为了评价PAF-R信号传导在黑素瘤细胞生长中的重要性，用含有PAF-R的cDNA的逆转录病毒转导PAF-R阴性鼠B16 F10细胞以产生稳定表达PAF-R的细胞（B16-PAF-R）或空载体（MSCV）以产生PAF-R缺陷型B16-MSCV对照细胞。与B16-MSCV细胞相比，使用PAF-R激动剂1-十六烷基-2-N-甲基氨基甲酰基-3-甘油磷酸胆碱激活PAF-R可诱导B16-PAF-R细胞增殖增加。逆转录定量聚合酶链反应显示，在人黑色素瘤SK23 MEL细胞中存在功能性PAF-R，但在SK 5 MEL细胞中不存在。本研究调查了在存在或不存在功能性PAF-R的情况下，BITC治疗对鼠和人黑素瘤细胞存活的影响。结果显示，用BITC处理降低了PAF-R阳性和阴性鼠和人黑素瘤细胞的存活率。然而，与PAF-R阴性细胞相比，在较低浓度下，PAF-R阳性细胞中PAF-R的表达显著增强了BITC介导的细胞毒性。为了确定潜在的机制，使用流式细胞术分析，其证明与B16-MSCV细胞相比，B16-PAF-R细胞中活性氧（ROS）的产生显著增加，这增强了BITC的凋亡，如通过增加的半胱天冬酶-3/7发光所测量的。值得注意的是，B16-PAF-R细胞中BITC介导的细胞存活率降低、ROS增加和凋亡增加被抗氧化剂维生素C显著减弱，表明ROS参与。此外，WEB2086 PAF-R拮抗剂抑制B16-PAF-R细胞中BITC介导的凋亡增强，表明PAF-R信号传导在BITC介导的效应中的作用。这些发现表明BITC对黑色素瘤中的PAF-R的选择性为PAF-R阳性黑色素瘤治疗提供了有希望的化学预防剂。
Melanoma cells often express platelet-activating factor receptor (PAF-R), which has been demonstrated to increase metastatic behavior. However, the effect of PAF-R on the responsiveness of melanoma to naturally occurring cytotoxic agents remains to be elucidated. The present study aimed to determine the relative cytotoxicity and mechanism of benzyl isothiocyanate (BITC), a component of cruciferous vegetables, in melanoma cells expressing PAF-R. To evaluate the importance of PAF-R signaling in melanoma cell growth, PAF-R-negative murine B16F10 cells were transduced with a retrovirus containing the cDNA for PAF-R to generate cells stably expressing PAF-R (B16-PAF-R) or an empty vector (MSCV) to generate PAF-R-deficient B16-MSCV control cells. Activation of PAF-R, using the PAF-R agonist, 1-hexadecyl-2-N-methylcarbamoyl-3-glycerophosphocholine, induced an increase in the proliferation of B16-PAF-R cells compared with the B16-MSCV cells. Reverse transcription quantitative polymerase chain reaction revealed the presence of functional PAF-R in human melanoma SK23MEL cells, but not in SK5MEL cells. The present study investigated the effect of BITC treatments on the survival of murine and human melanoma cells, in the presence or absence of functional PAF-R. The results revealed that treatment with BITC decreased the survival rate of the PAF-R-positive and negative murine and human melanoma cells. However, the expression of PAF-R substantially augmented BITC-mediated cytotoxicity in the PAF-R-positive cells at lower concentrations compared with the PAF-R-negative cells. In order to determine the underlying mechanism, flow cytometric analysis was used, which demonstrated a significant increase in the generation of reactive oxygen species (ROS) in the B16-PAF-R cells compared with the B16-MSCV cells, which enhanced apoptosis by BITC, as measured by increased caspase-3/7 luminescence. Notably, the BITC-mediated decreased cell survival rate, increased ROS and increased apoptosis in the B16-PAF-R cells were significantly attenuated by the antioxidant, vitamin C, indicating ROS involvement. Additionally, the WEB2086 PAF-R antagonist, inhibited the BITC-mediated enhancement of apoptosis in the B16-PAF-R cells, indicating a role for PAF-R-signaling in the BITC-mediated effects. These findings indicated that the selectivity of BITC towards PAF-R in melanoma offers a promising chemopreventive agent for PAF-R-positive melanoma treatment.
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影响因子：
3.8
作者：
de Oliveira, Soraya I.;Andrade, Luciana N. S.;Jancar, Sonia
通讯作者：
Jancar, Sonia
影响因子：
4.8
作者：
Li, T;Southall, MD;Travers, JB
通讯作者：
Travers, JB
影响因子：
4.7
作者：
Sahu, Ravi P.;Kozman, Amal A.;Konger, Raymond L.
通讯作者：
Konger, Raymond L.
影响因子：
10.3
作者：
Sahu, Ravi P.;Srivastava, Sanjay K.
通讯作者：
Srivastava, Sanjay K.
影响因子：
9.7
作者：
Stoner, GD;Morse, MA
通讯作者：
Morse, MA