COL2A1 Is a Novel Biomarker of Melanoma Tumor Repopulating Cells.
COL2A1 Is a Novel Biomarker of Melanoma Tumor Repopulating Cells.
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DOI：
10.3390/biomedicines8090360
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发表时间：
2020-09-18
期刊：
影响因子：
4.7
通讯作者：
Chowdhury F
中科院分区：
文献类型：
--
作者：
Talluri B;Amar K;Saul M;Shireen T;Konjufca V;Ma J;Ha T;Chowdhury F
从B16 F1黑色素瘤细胞系中选择的软3D纤维蛋白凝胶肿瘤再生细胞（TRC）表现出非凡的自我更新和肿瘤再生能力。然而，它们的生物标志物和基因调控特征在很大程度上仍然未知。在这里，我们利用下一代基于测序的RNA测序（RNA-seq）技术来发现TRC的新生物标志物和活性基因调控特征。RNA-seq数据的系统生物学分析鉴定了差异表达的基因簇，包括细胞粘附簇，随后鉴定了高度特异性和新型生物标志物，如Col 2a 1，Ncam 1，F11 r和Negr 1。我们通过实时qPCR验证了这些基因的表达。发现Col 2a 1的表达水平在TRC中相对较低，但与亲本对照细胞系相比高20倍，因此使得生物标志物对TRC非常特异。我们通过免疫荧光显微镜验证了COL 2A 1蛋白，显示与亲本对照细胞相比，COL 2A 1在TRC中的表达更高。KEGG通路分析显示JAK/STAT、缺氧和Akt信号通路在TRCs中是活跃的。此外，有氧糖酵解途径非常活跃，表明高致瘤性细胞具有典型的瓦尔堡效应。总之，我们的研究揭示了黑色素瘤TRC的高度特异性生物标志物和活性细胞信号传导途径，这些生物标志物和活性细胞信号传导途径可以潜在地靶向和中和TRC。
Soft 3D-fibrin-gel selected tumor repopulating cells (TRCs) from the B16F1 melanoma cell line exhibit extraordinary self-renewal and tumor-regeneration capabilities. However, their biomarkers and gene regulatory features remain largely unknown. Here, we utilized the next-generation sequencing-based RNA sequencing (RNA-seq) technique to discover novel biomarkers and active gene regulatory features of TRCs. Systems biology analysis of RNA-seq data identified differentially expressed gene clusters, including the cell adhesion cluster, which subsequently identified highly specific and novel biomarkers, such as Col2a1, Ncam1, F11r, and Negr1. We validated the expression of these genes by real-time qPCR. The expression level of Col2a1 was found to be relatively low in TRCs but twenty-fold higher compared to the parental control cell line, thus making the biomarker very specific for TRCs. We validated the COL2A1 protein by immunofluorescence microscopy, showing a higher expression of COL2A1 in TRCs compared to parental control cells. KEGG pathway analysis showed the JAK/STAT, hypoxia, and Akt signaling pathways to be active in TRCs. Besides, the aerobic glycolysis pathway was found to be very active, indicating a typical Warburg Effect on highly tumorigenic cells. Together, our study revealed highly specific biomarkers and active cell signaling pathways of melanoma TRCs that can potentially target and neutralize TRCs.
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影响因子：
13.8
作者：
Liberti MV;Locasale JW
通讯作者：
Locasale JW
影响因子：
4.6
作者：
Chen J;Zhou W;Jia Q;Chen J;Zhang S;Yao W;Wei F;Zhang Y;Yang F;Huang W;Zhang Y;Zhang H;Zhang Y;Huang B;Zhang Z;Jia H;Wang N
通讯作者：
Wang N
影响因子：
3.7
作者：
Chowdhury F;Li Y;Poh YC;Yokohama-Tamaki T;Wang N;Tanaka TS
通讯作者：
Tanaka TS
影响因子：
6.2
作者：
Jia Q;Zhou W;Yao W;Yang F;Zhang S;Singh R;Chen J;Chen JJ;Zhang Y;Wei F;Zhang Y;Jia H;Wang N
通讯作者：
Wang N
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y