Onconase mediated NFKβ downregulation in malignant pleural mesothelioma.
Onconase mediated NFKβ downregulation in malignant pleural mesothelioma.
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DOI：
10.1038/onc.2010.643
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发表时间：
2011-06-16
期刊：
影响因子：
8
通讯作者：
Pass, H. I.
中科院分区：
文献类型：
--
作者：
Goparaju, C. M.;Blasberg, J. D.;Volinia, S.;Palatini, J.;Ivanov, S.;Donington, J. S.;Croce, C.;Carbone, M.;Yang, H.;Pass, H. I.
用Ranpirnase （Onconase）治疗恶性胸膜间皮瘤（MPM）会导致蛋白质翻译的破坏和细胞凋亡。我们假设Onconase通过特异性microRNAs （miRNA）下调核因子κ B （NFKβ）而起作用，并且该途径的干扰可能与MPM对化疗的耐药有关。三种永生化MPM细胞系（H2959， H2373和H2591）暴露于0-20µg/mL的Onconase中。在48小时和72小时测量细胞计数。RT-PCR验证mirna富集RNA中的基因表达。通过将miRNA模拟物或抑制剂转染到MPM细胞系中，并进行Matrigel™侵袭、细胞增殖、软琼脂集落形成和划痕闭合试验，评估miRNA表达的功能意义。通过RT-PCR和Western Blotting评估NFKβ表达和下游靶标包括ABC转运蛋白、BCL-xl和IAP的影响。20µg/mL Onconase处理显著降低细胞计数和侵袭。在所有细胞系中，经Onconase处理，Hsa-miR-17*显著上调，hsa-miR-30c显著下调。在所有实验中，强迫表达hsa-miR-17* mimic和hsa-miR-30c inhibitor均显著降低Onconase的功能活性。NFKB1（p50）表达和下游靶标也在Onconase治疗以及强迫表达miRNA模拟物和抑制剂的情况下降低。Onconase处理导致细胞增殖、侵袭和某些mirna表达显著降低。用hsa-miR-17*模拟物和hsa-miR-30c抑制剂重述所得到的miRNA表达模式，在功能分析中导致NFKB1的下调和恶性行为的减少。因此，Onconase可能通过这些mirna发挥其抗肿瘤作用。
Treatment of malignant pleural mesothelioma (MPM) with Ranpirnase (Onconase) results in disruption of protein translation and cell apoptosis. We hypothesize that Onconase acts via down regulation of nuclear factor kappa B (NFKβ) by specific microRNAs (miRNA) and that interference of this pathway could have implications for MPM resistance to chemotherapy. Three immortalized MPM cell lines (H2959, H2373, and H2591) were exposed to Onconase at 0–20 µg/mL. Cell counts were measured at 48 and 72 hours. Gene expression in miRNA-enriched RNA was validated by RT-PCR. The functional implications of miRNA expression were evaluated by transfecting miRNA mimics or inhibitors into MPM cell lines, and performing Matrigel™ invasion, cell proliferation, soft agar colony formation, and scratch closure assays. Effects on NFKβ expression and downstream targets including ABC transporters, BCL-xl, and IAP were assessed by RT-PCR and Western Blotting. Treatment with 20µg/mL of Onconase significantly decreased cell count and invasion. Hsa-miR-17* was significantly upregulated and hsa-miR-30c significantly down-regulated by Onconase treatment in all cell lines. Forced expression of hsa-miR-17* mimic and hsa-miR-30c inhibitor each significantly decreased functional activity of Onconase in all assays. NFKB1(p50) expression and downstream targets were also decreased with Onconase treatment as well as with forced expression miRNA mimic and inhibitors. Onconase treatment caused a significant decrease in cell proliferation, invasion, and in expression of certain miRNAs. Recapitulation of the resultant miRNA expression pattern with hsa-miR-17* mimic and hsa-miR-30c inhibitor resulted in downregulation of NFKB1 and reduced malignant behavior in functional assays. Thus, Onconase likely exerts its anti-tumor effect through these miRNAs.
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影响因子：
14.9
作者：
Chen C;Ridzon DA;Broomer AJ;Zhou Z;Lee DH;Nguyen JT;Barbisin M;Xu NL;Mahuvakar VR;Andersen MR;Lao KQ;Livak KJ;Guegler KJ
通讯作者：
Guegler KJ
影响因子：
3.3
作者：
Kbeck, Amanda;Pass, Harvey I.;Yang, Haining
通讯作者：
Yang, Haining
影响因子：
5.6
作者：
Kobe, B;Deisenhofer, J
通讯作者：
Deisenhofer, J
影响因子：
3.6
作者：
Cao, Xiaobo;Rodarte, Charles;Smythe, W. Roy
通讯作者：
Smythe, W. Roy
影响因子：
4.3
作者：
Ardelt, Barbara;Ardelt, Wojciech;Darzynkiewicz, Zbigniew
通讯作者：
Darzynkiewicz, Zbigniew