Overexpression of miR-15b Promotes Resistance to Sunitinib in Renal Cell Carcinoma
Overexpression of miR-15b Promotes Resistance to Sunitinib in Renal Cell Carcinoma
复制标题
miR-15b 的过度表达促进肾细胞癌对舒尼替尼的耐药性
DOI：
10.7150/jca.31676
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发表时间：
2019-01-01
影响因子：
3.9
通讯作者：
Feng, Chenchen
中科院分区：
文献类型：
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作者：
Lu, Lu;Li, Yanmin;Feng, Chenchen
舒尼替尼仍然是转移性透明细胞肾细胞癌（ccRCC）的一线治疗。耐药是不可避免的，相关机制值得深入探讨。方法：对GEO和TCGA数据集进行计算机数据挖掘，以识别潜在的靶micro-RNA。进行体外和体内研究以验证结果。结果：GEO数据集的再现显示miR-15 b在舒尼替尼耐药的ccRCC中显著上调。在舒尼替尼治疗后，7种ccRCC细胞系中有5种表现出显著过表达miR-15 b。载体介导的miR-15 b过表达显著诱导ccRCC细胞对舒尼替尼的耐药性。过表达miR-15 b可显著减少舒尼替尼处理的细胞G1期细胞数和细胞凋亡。将TCGA ccRCC（KIRC）数据集中与miR-15 b负相关的基因表达与miR-15 b的预测靶标交叉参考，并选择CCNC作为耐药介导的潜在靶标。过表达miR-15 b抑制CCNC表达和蛋白质（细胞周期蛋白C）水平。细胞周期蛋白C相关蛋白CDK 19和CDK 8也在miR-15 b过表达后受到抑制。沉默CCNC模拟miR-25的过表达，诱导细胞周期进展通过G1期和舒尼替尼处理的ccRCC细胞中较少的凋亡。miR-15 b的过表达也抵消了舒尼替尼对ccRCC细胞系中迁移和集落形成的抑制。体内小鼠异种移植模型显示，在舒尼替尼治疗的小鼠中，miR-15 b表达恢复了肿瘤生长。结论：我们在这里显示miR-15 b是ccRCC中舒尼替尼耐药的可能罪魁祸首。靶向miR-15 b可能克服耐药性，相关机制值得进一步研究。
Aim: Sunitinib remains the frontline treatment for metastatic clear-cell renal cell carcinoma (ccRCC). Drug resistance is inevitable and related mechanism warrant insightful elaboration. Methods: In silico data mining of GEO and TCGA datasets was performed to identify potential target micro-RNA. In vitro and in vivo studies were performed to validate findings. Results: Reproduction of GEO datasets revealed miR-15b significantly upregulated in sunitinib- resistant ccRCC. Five out of seven ccRCC cell lines demonstrated significantly overexpressed miR-15b after sunitinib treatment. Vector-mediated overexpression of miR-15b significantly induced resistance to sunitinib in ccRCC cells. Overexpression of miR-15b significantly induced less population in G1 phase of cell cycle and less apoptosis in cells treated sunitinib. Expression of genes negatively correlated with miR-15b in TCGA ccRCC (KIRC) dataset were cross-referenced with predicted targets of miR-15b and CCNC was selected as potential target for resistance mediation. Overexpression of miR-15b suppressed CCNC expression and protein (Cyclin C) levels. Cyclin C-associated proteins CDK19 and CDK8 were also suppressed following miR-15b overexpression. Silencing of CCNC mimicked overexpression of miR-25 inducing cell cycle progression passing G1 phase and less apoptosis in ccRCC cells treated by sunitinib. Overexpression of miR-15b also counteracted suppression of migration and colony formation by sunitinib in ccRCC cell lines. In vivo mouse xenograft models showed recovered tumor growth with miR-15b expression in mice treated with sunitinib. Conclusion: We here show miR-15b as a possible culprit for sunitinib resistance in ccRCC. Targeting miR-15b could potentially overcome drug resistance and related mechanism warrants further investigation.