癌症干细胞PCR芯片 Cancer Stem Cells

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Cancer Stem Cells PCR Array

癌症干细胞PCR芯片

Product	Species	Technology	Cat. No.
Cancer Stem Cells PCR Array	Human	Gene Expression	PAHS-176Z
Cancer Stem Cells PCR Array	Mouse	Gene Expression	PAMM-176Z
Cancer Stem Cells PCR Array	Rat	Gene Expression	PARN-176Z

The Human Cancer Stem Cells RT2 Profiler PCR array profiles the expression of 84 genes linked to cancer stem cells (CSCs). Cancer researchers have struggled with the vexing problem that although many cancer drugs dramatically reduce the size of the tumors, most cancers eventually relapse. Dynamic changes in cancer cell populations during treatment suggest that a small population of cells resistant to current therapies is ultimately responsible for the re-growth of tumors.Furthermore, studies imply that these cells may provide a reservoir for the generation and propagation of mutant cells providing further resistance to therapy. The cancer-stem-cell hypothesis posits that only a very rare population of cells within tumors has the capacity for limitless self-renewal.This concept has important therapeutic implications, and may explain why many cancers return even after treatment removes any detectable tumor cells.If current treatments do not eliminate cancer stem cells, then they may regenerate the tumor once treatment stops. Recently, advances in technology have allowed the prospective identification and purification of CSCs from various different types of cancers for further characterization. The genes profiled with this array include CSC molecular markers and genes regulating CSC proliferation, self-renewal, and pluripotency to help ensure the stability of CSC isolates in culture. Also included are genes involved in CSC asymmetric cell division, migration and metastasis, and relevant signal transduction pathways to help facilitate CSC characterization as well as the targets of therapeutics currently being tested. A set of controls present on each array enables data analysis using the ΔΔCT method of relative quantification and assessment of reverse transcription performance, genomic DNA contamination, and PCR performance. Using real-time PCR, research studies can easily and reliably analyze the expression of a focused panel of genes related to cancer stem cells with this array.
The RT² Profiler PCR Arrays are intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
癌症干细胞PCR芯片用于研究与肿瘤干细胞(CSCs)相关的84个基因的表达。癌症研究人员疲于应对棘手的问题,尽管许多抗癌药物大幅减少肿瘤的大小,最终大多数癌症复发。在治疗期间癌症细胞种群的动态变化表明,一个小种群细胞抵抗目前的治疗方法是最终负责肿瘤复发。此外,研究表明,这些细胞可能会提供一个突变细胞的生成和传播储蓄池提供进一步的抵抗治疗。肿瘤干细胞假说认为在肿瘤中只有很少的细胞具有无限的自我更新能力。这一概念具有重要的治疗意义,甚至也许可以解释为什么许多癌症恢复甚至在治疗清除任何检测到的肿瘤细胞后。目前的治疗如果不消除癌症干细胞,那么一旦停止治疗它们可能会再生肿瘤。最近技术的进步使得CSCs从不同类型的癌症中做的预期鉴定和纯化作进一步鉴定。这个芯片包括CSC分子标记和调节CSC增殖、自我更新和多潜能帮助确保CSC隔离培养的稳定性的基因。还包括参与CSC不对称细胞分裂、迁移和转移、促进CSC鉴定的相关信号转导通路的基因，以及当前已测试的治疗靶点。每张芯片含一个对照组使得分析数据时可以用相对定量ΔΔCT方法评估逆转录效率,基因组DNA污染,和PCR效率。利用这张芯片，通过实时定量PCR，可以简易且可靠地分析癌症干细胞相关基因集的表达。

PCR芯片仅用于分子生物学应用。本产品不用于疾病的诊断、预防和治疗。

Cancer Stem Cell Markers: ABCB5, ALCAM, ALDH1A1, ATXN1, BMI1, CD24, CD34, CD38, CD44, ENG, ETFA, FLOT2, GATA3, ITGA2, ITGA4, ITGA6, ITGB1, KIT, MS4A1, MUC1, PECAM1, PROM1, PTPRC, THY1.
Proliferation: EGF, ERBB2, KITLG, LIN28B, NOS2.
Self-Renewal: BMP7, DNMT1, FGFR2.
Pluripotency: KLF4, LIN28A, MYC, NANOG, POU5F1, SOX2.
Asymmetric Division: FOXP1, HDAC1, MYCN, SIRT1, WNT1.
Migration & Metastasis: AXL, ID1, IL8, KLF17, PLAT, PLAUR, SNAi1, TWIST1, TWIST2, ZEB1, ZEB2.
Loss of Stemness: ALDH1A1, CD34, DACH1, FOXA2, PECAM1, PTCH1.
Signal Transduction Pathways:
Hippo Signaling: LATS1, MERTK, SAV1, TAZ, WWC1, YAP1.
Hedgehog Signaling: PTCH1, SMO.
Notch Signaling: DLL1, DLL4, JAG1, MAML1, NOTCH1, NOTCH2.
WNT Signaling: DKK1, EPCAM, FZD7, WNT1.
PI3K/AKT/mTOR signaling: ABCG2, GSK3B.
STAT/NFκB Signaling: IKBKB, JAK2, NFKB1.
Therapeutic Targets: ABCG2, ATM, AXL, CHEK1, DDR1, DKK1, EPCAM, FZD7, GSK3B, ID1, IKBKB, JAK2, KLF17, NFKB1, PTCH1, SMO, STAT3, TGFBR1, WEE1.

How it Works

The PCR array is a set of optimized real-time PCR primer assays on 96-well or 384-well plates for pathway or disease focused genes as well as appropriate RNA quality controls. The PCR array performs gene expression analysis with real-time PCR sensitivity and the multi-gene profiling capability of a microarray. Simply mix your cDNA template with the appropriate ready-to-use PCR master mix, aliquot equal volumes to each well of the same plate, and then run the real-time PCR cycling program. (Download user manual)

Figure 1:	How PCR Arrays Work - Protocol Chart

What it offers?
Guaranteed Performance* - ready-to-use for gene expression analysis
Time and cost saving - less than 30 min hands-on time for analyzing 84 genes
Ease of data analysis using our easy-to-use Excel-based data analysis template or web-based analysis tool

Layout and Controls: The PCR Arrays are available in both 96- and 384-well plates and are used to monitor the expression of 84 genes related to a disease state or pathway plus five housekeeping genes. Controls are also included on each array for genomic DNA contamination, RNA quality, and general PCR performance

You can easily perform a PCR Array experiment in your own laboratory, or send your samples to us and take advantage of our PCR Array Services.

*: when using complete PCR array system.

Performance Data Sensitivity:
The complete PCR Array System yields a greater-than 85 percent present call with as little as 25 ng as much as 5 µg of total RNA from a pathway representing genes expressed at a lower level (inflammatory cytokines and receptors).

Figure 2:	PCR Arrays Let You See More Genes with Less RNA Different amounts of universal total RNA were characterized using the Human Inflammatory Cytokines and Receptors PCR Array, and the percentage of detectable genes was calculated for each RNA amount. As little as 25 ng total RNA yields greater than an 80% positive call, even for cytokines expressed at very low levels.

Reproducibility
The complete PCR Array System demonstrates strong correlations across technical replicates, lots, and instruments with average correlation coefficients > 0.99 insuring reliable detection of differences in expression between biological samples.

	050825_1	050825_2	050825_3	050825_4
060111_1	0.993	0.989	0.995	0.992
060111_2	0.994	0.990	0.995	0.992
060111_3	0.992	0.990	0.993	0.992
060111_4	0.993	0.992	0.994	0.992

Figure 3:	PCR Arrays Yield Highly Reproducible Results Four replicate sets of raw threshold data (1-4) obtained by two different scientists (A & B) at two different times (050825 & 060111) on Human Drug Metabolism RT² Profiler PCR Arrays are directly compared. The results demonstrate a high degree of correlation (R2 > 0.990).

Specificity
The complete PCR Array System, with high quality input RNA, is guaranteed to yield single bands of the predicted size without primer dimers or other secondary products thus providing the most accurate real-time PCR results possible.

Figure 4:	PCR Arrays Amplify A Single Gene-Specific Product in Every Reaction. Universal total RNA was characterized on the TGFβ / BMP Signaling Pathway PCR Array, followed by dissociation (melt) curve analysis. PCR Arrays specifically detect individual genes despite the expression of related gene family members in the same RNA sample.

Application Data

Cancer Research:
To ascertain the oncogenic route that two different human breast tumors have taken, the relative expression level of cancer- and adhesion-related genes in normal and two different cancerous tissues were compared.

Template cDNAs prepared from total RNA of normal human breast and two human breast tumors (BioChain Institute, Inc., 5.0 µg) were characterized in technical triplicates using the Human Cancer PathwayFinder™ PCR Array and the Human Extracellular Matrix & Adhesion Molecule PCR Array with the RT² SYBR Green / Fluorescein PCR master mix on the Bio-Rad iCycler®.

Figure 5:

ECM & Adhesion PCR Arrays Revealed Up- and Down-Regulated Genes in Breast Cancer Total RNA from normal human breast and a human breast tumor were characterized in technical triplicates, and the relative expression levels for each gene in the two samples are plotted against each other in the Scatter Plot. Genes encoding the matrix metallopeptidases (MMP3 & MMP9) and their inhibitors (TIMP3) are up-regulated, while genes encoding integrins (ITGB3 & ITGB4) are down-regulated, by at least three-fold (outside the silver field) in breast tumors relative to normal tissue.

Toxicology Research:
Rezulin (Troglitazone or "Tro" or "T"), a glitazone PPAR-gamma agonist, was approved for treatment of type 2 diabetes mellitus, but was withdrawn from the market due to idiosyncratic liver toxicity. Two similar drugs, Avandia (Rosiglitazone or "Rosi" or "R") and Actos (Pioglitazone or "Pio" or "P"), are considered to be safe treatments for the same condition. The expression profile of key drug metabolism genes should be different in cells treated with Rezulin versus those treated with Avandia and Actos.

Hepatocellular carcinoma HepG2 cells were treated at 80% cell confluence with these three drugs (100 µM, Cayman Chemical) or a DMSO vehicle control for 24 h. RNA isolated using the ArrayGrade™ Total RNA Isolation Kit was used to characterize gene expression with the Human Drug Metabolism and Stress & Toxicity PathwayFinder™ RT² Profiler™ PCR Arrays and RT² SYBR Green / Fluorescein PCR master mix on the Bio-Rad iCycler®.

Figure 6:

Stress & Toxicity PathwayFinder™ PCR Array Uncovered Idiosyncratic Mechanisms of Action for Liver Toxicity Caused by 3 PPARγ Agonists. RNA from HepG2 cells treated with three different glitazone PPARγ agonists for type 2 diabetes mellitus was characterized, and the results were compared to that of a vehicle (DMSO) control. A withdrawn drug with idiosyncratic liver toxicity (Rezulin) induces very different changes in the expression of stress-related genes than two safer drugs still on the market (Avandia and Actos).