Three dysregulated miRNAs control kallikrein 10 expression and cell proliferation in ovarian cancer.
Three dysregulated miRNAs control kallikrein 10 expression and cell proliferation in ovarian cancer.
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DOI：
10.1038/sj.bjc.6605634
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发表时间：
2010-04-13
影响因子：
8.8
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中科院分区：
文献类型：
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激肽释放酶相关肽酶（KLK）是丝氨酸蛋白酶家族，其已被证明在包括卵巢癌在内的几种恶性肿瘤中失调。激肽释放酶基因的控制及其在癌症中的生理功能还不清楚。我们假设microRNAs（miRNAs）代表了一种新的转录后控制癌症中KLK表达的机制。我们首先在计算机上分析了卵巢癌中的miRNA表达。据报道，共有98种miRNAs在卵巢癌中表达改变。这些miRNAs中的三种被预测靶向KLK 10。我们使用两种独立的技术实验验证了预测的miR-KLK 10相互作用，一种是使用含有KLK 10 3′非翻译区（UTR）的构建体pMIR-KLK 10的荧光素酶测定，另一种是测量用miRNA转染后的KLK 10蛋白水平。当我们用pMIR-KLK 10和let-7 f、miR-224或mR-516 a共转染细胞时，我们看到荧光素酶信号减少，表明这些miRNA可以靶向KLK 10。然后，我们研究了这三种miRNA对KLK 10蛋白表达和细胞生长的影响。转染所有miRNA，let-7 f，miR-224和miR-516 a导致蛋白质表达和细胞生长的降低。该效应具有剂量依赖性。共转染let-7 f及其抑制剂可部分恢复KLK 10蛋白水平。另外，let-7 f转染后，KLK 10 mRNA表达略有下降。我们的研究结果证实，KLK可以被一个以上的miRNA靶向。卵巢癌中某些miRNAs的表达增加可导致KLK蛋白表达降低，随后对细胞增殖产生负面影响。这种剂量依赖性效应表明存在一种“微调”或“微调”机制，其中一种KLK的表达可以由多种miRNA控制。这些数据共同表明，miRNA可能被用作潜在的治疗选择，需要进一步的研究。
Kallikrein-related peptidases (KLKs) are a family of serine proteases that have been shown to be dysregulated in several malignancies including ovarian cancer. The control of kallikrein genes and their physiological function in cancer is not well understood. We hypothesized that microRNAs (miRNAs) represent a novel mechanism for post-transcriptional control of KLK expression in cancer. We first analysed miRNA expression in ovarian cancer in silico. A total of 98 miRNAs were reported to have altered expression in ovarian cancer. Three of these miRNAs were predicted to target KLK10. We experimentally verified the predicted miR–KLK10 interaction using two independent techniques, a luciferase assay with a construct containing the KLK10 3′ untranslated region (UTR), pMIR–KLK10, and measuring KLK10 protein levels after transfection with miRNA. When we co-transfected cells with pMIR–KLK10 and either let-7f, miR-224, or mR-516a, we saw decreased luciferase signal, suggesting that these miRNAs can target KLK10. We then examined the effect of these three miRNAs on KLK10 protein expression and cell growth. Transfection of all miRNAs, let-7f, miR-224, and miR-516a led to a decrease in protein expression and cellular growth. This effect was shown to be dose dependent. The KLK10 protein levels were partially restored by co-transfecting let-7f and its inhibitor. In addition, there was a slight decrease in KLK10 mRNA expression after transfection with let-7f. Our results confirm that KLKs can be targeted by more than one miRNA. Increased expression of certain miRNAs in ovarian cancer can lead to decreased KLK protein expression and subsequently have a negative effect on cell proliferation. This dose-dependent effect suggests that a ‘tweaking’ or ‘fine-tuning’ mechanism exists in which the expression of one KLK can be controlled by multiple miRNAs. These data together suggest that miRNA may be used as potential therapeutic options and further studies are required.
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影响因子：
11.5
作者：
Dorn, Julia;Schmitt, Manfred;Harbeck, Nadia
通讯作者：
Harbeck, Nadia
影响因子：
5
作者：
Emami, Nashmil;Diamandis, Eleftherios P.
通讯作者：
Diamandis, Eleftherios P.
影响因子：
45.3
作者：
Diamandis, EP;Scorilas, A;Katsaros, D
通讯作者：
Katsaros, D
影响因子：
6.6
作者：
Bayani, Jane;Paliouras, Miltiadis;Diamandis, Eleftherios P.
通讯作者：
Diamandis, Eleftherios P.
影响因子：
--
作者：
Arora A;Guduric-Fuchs J;Harwood L;Dellett M;Cogliati T;Simpson DA
通讯作者：
Simpson DA