BA/F3_EGFR Del19-T790M激酶Kinase稳定表达细胞株-BioVector NTCC保藏中心

BA/F3_EGFR Del19-T790M激酶Kinase稳定表达细胞株-BioVector NTCC保藏中心
I. Introduction
Cell Line Name：

EGFR-d19/T790M

Gene Synonyms:

Epidermal Growth Factor Receptor, ERBB1, EGFR

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

Mycoplasma Status:

Negative


II.Background
EGFR is widely recognized for its importance in cancer. Amplification and mutations have been shown to be driving events in many cancer types. Its role in non-small cell lung cancer, glioblastoma and basal-like breast cancers has spurred many research and drug development efforts. Tyrosine kinase inhibitors have shown efficacy in EGFR amplfied tumors, most notably gefitinib and erlotinib. Mutations in EGFR have been shown to confer resistance to these drugs, particularly the variant T790M, which has been functionally characterized as a resistance marker for both of these drugs. The later generation TKI's have seen some success in treating these resistant cases, and targeted sequencing of the EGFR locus has become a common practice in treatment of non-small cell lung cancer. Overproduction of ligands is another possible mechanism of activation of EGFR. ERBB ligands include EGF, TGF-a, AREG, EPG, BTC, HB-EGF, EPR and NRG1-4 (for detailed information please refer to the respective ligand section). In ligand-activated cancers, Cetuximab appears to be more effective than tyrosine-kinase inhibitors.


III. Representative Data
1. Anti-proliferation assay



Figure 3. Anti-proliferation assay of three reference compounds on the Ba/F3_EGFR-d19/T790M 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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