BA/F3_ETV6-NTRK3激酶Kinase稳定表达细胞株-BioVector NTCC保藏中心

BA/F3_ETV6-NTRK3激酶Kinase稳定表达细胞株-BioVector NTCC保藏中心
I. Introduction
Cell Line Name：

BA/F3_ETV6-NTRK3

Gene Synonyms:

ETS variant 6 ,TEL, TEL/ABL, THC5;neurotrophic receptor tyrosine kinase 3 ,GP145-TrkC, TRKC, gp145(trkC)

Host Cell:

Ba/F3

Stability: 16 passages
Application:

Anti-proliferation assay and PD assay

Freeze Medium:

90% FBS+10% DMSO

Complete Culture Medium:

RPMI-1640+10%FBS+1 ug/ml puromycin

Mycoplasma Status:

Negative


II.Background
ETV6-NTRK3 gene fusion is the translocation of genetic material between the ETV6 gene located on the short arm (designated p) of chromosome 12 at position p13.2 (i.e. 12p13.2) and the NTRK3 gene located on the long arm (designated q) of chromosome 15 at position q25.3 (i.e. 15q25.3) to create the (12;15)(p13;q25) fusion gene, ETV6-NTRK3. This new gene consists of the 5' end of ETV6 fused to the 3' end of NTRK3. ETV6-NTRK3 therefore codes for an chimeric oncoprotein consisting of the helix-loop-helix (HLH) protein dimerization domain of the ETV6 protein fused to the tyrosine kinase (i.e. PTK) domain of the NTRK3 protein. The ETV6 gene codes for the transcription factor protein, ETV6, which suppresses the expression of, and thereby regulates, various genes that in mice are required for normal hematopoiesis as well as the development and maintenance of the vascular network. NTRK3 codes for Tropomyosin receptor kinase C a NT-3 growth factor receptor cell surface protein that when bound to its growth factor ligand, neurotrophin-3, becomes an active tyrosine kinase that phosphorylates tyrosine residues on, and thereby stimulates, signaling proteins that promote the growth, survival, and proliferation of their parent cells. The tyrosine kinase of the ETV6-NTRK3 fusion protein is dysfunctional in that it is continuously active in phophorylating tyrosine residues on, and thereby continuously stimulating, proteins that promote the growth, survival, and proliferation of their parent cells. In consequence, these cells take on malignant characteristics and are on the pathway of becoming cancerous. Indeed, the ETV6-NTRK3 fusion gene appears to be a critical driver of several types of cancers. It was originally identified in congenital fibrosarcoma and subsequently found in secretory breast cancer (also termed juvenile breast cancer), Mammary analogue secretory carcinoma of salivary glands (also termed MASC or MASCSG), congenital fibrosarcoma, congenital mesoblastic nephroma, rare cases of acute myelogenous leukemia, ALK-negative Inflammatory myofibroblastic tumour, and radiation-induced papillary thyroid carcinoma.


III. Representative Data
1. Anti-proliferation assay



Figure 1. Anti-proliferation assay of two reference compounds on the BA/F3_ETV6-NTRK3 Stable Cell Line


IV. Thawing

Thawing: Protocol
1. Remove the vial from liquid nitrogen tank and thaw cells quickly in a 37°C water-bath.
2. Just before the cells are completely thawed, decontaminate the outside of the vial with 70% ethanol and transfer the cells to a 15 ml centrifuge tube containing 9 ml of complete growth medium.
3. Pellet cells by centrifugation at 200 x g force for 5 min, and discard the medium.
4. Resuspend the cells in complete growth medium.
5. Add 10 ml of the cell suspension in a 10 cm dish.
6. Add promycin to a concentration of 1 μg/ml the following day.
Kinase细胞株



现代新药研发的关键首先是寻找，确定和制备药物作用靶点。在500多个已发现的药物靶点里中，GPCR，Ion Channel，Kinase使用的最为广泛。




激酶(kinase)是一类从高能供体分子（如ATP）转移磷酸基团到特定靶分子（底物）的酶；这一过程谓之磷酸化。许多肿瘤的发生是由某些与生长相关的“激酶”发生突变导致异常活化引起的，因而针对这些突变激酶的抑制剂能够有效抑制这些激酶的活性，从而达到抑制癌细胞增长的目的。
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