PKC-alpha modulation by miR-483-3p in platinum-resistant ovarian carcinoma cells
PKC-alpha modulation by miR-483-3p in platinum-resistant ovarian carcinoma cells
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DOI：
10.1016/j.taap.2016.08.005
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发表时间：
2016-11-01
影响因子：
3.8
通讯作者：
Gatti, Laura
中科院分区：
文献类型：
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作者：
Arrighetti, Noemi;Cossa, Giacomo;Gatti, Laura
耐药的发生限制了铂类化合物治疗卵巢癌的疗效。由于microRNAs （miRNAs）可能通过调节肿瘤细胞反应的不同方面来促进这种现象，因此本研究的目的是利用miRNAs在铂敏感/耐药细胞中的表达分析，试图确定药物反应的潜在调节因子。MiR-483-3p可能参与细胞凋亡和细胞增殖调节，在4种铂耐药变体中，特别是在IGROV-1/Pt1亚系中，与亲代细胞相比，MiR-483-3p被发现上调。在IGROV-1亲本细胞中转染miR-483-3p的合成前体可引起miRNA水平的显著上调。生长抑制和集落形成实验表明，miR-483-3p过表达通过干扰IGROV-1细胞的增殖潜能，降低了细胞生长，并赋予IGROV-1细胞轻度的顺铂耐药性。miR-483-3p的预测靶标包括PRKCA（编码PKC-alpha），此前报道与卵巢癌铂耐药相关。我们发现miR-483-3p直接靶向IGROV-1细胞中的PRKCA。与这一发现一致，IGROV-1细胞对顺铂的敏感性在pkc - α的分子/药理学抑制下下降。总之，我们的研究结果表明，卵巢癌铂耐药细胞过表达miR-483-3p可能会干扰其增殖，从而保护其免受铂化合物诱导的DNA损伤，最终代表了一种耐药机制。细胞生长受损可能是低水平耐药的原因，这可能与临床环境有关。(C) 2016 Elsevier Inc.版权所有。
The occurrence of drug resistance limits the efficacy of platinum compounds in the cure of ovarian carcinoma. Since microRNAs (miRNAs) may contribute to this phenomenon by regulating different aspects of tumor cell response, the aim of this study was to exploit the analysis of expression of miRNAs in platinum sensitive/resistant cells in an attempt to identify potential regulators of drug response. MiR-483-3p, which may participate in apoptosis and cell proliferation regulation, was found up-regulated in 4 platinum resistant variants, particularly in the IGROV-1/Pt1 subline, versus parental cells. Transfection of a synthetic precursor of miR-483-3p in IGROV-1 parental cells elicited a marked up-regulation of the miRNA levels. Growth-inhibition and colony-forming assays indicated that miR-483-3p over-expression reduced cell growth and conferred mild levels of cisplatin resistance in IGROV-1 cells, by interference with their proliferative potential. Predicted targets of miR-483-3p included PRKCA (encoding PKC-alpha), previously reported to be associated to platinum-resistance in ovarian carcinoma. We found that miR-483-3p directly targeted PRKCA in IGROV-1 cells. In keeping with this finding, cisplatin sensitivity of IGROV-1 cells decreased upon molecular/pharmacological inhibition of PKC-alpha. Overall, our results suggest that overexpression of miR-483-3p by ovarian carcinoma platinum-resistant cells may interfere with their proliferation, thus protecting them from DNA damage induced by platinum compounds and ultimately representing a drug-resistance mechanism. The impairment of cell growth may account for low levels of drug resistance that could be relevant in the clinical setting. (C) 2016 Elsevier Inc. All rights reserved.