15-PGDH inhibits hepatocellular carcinoma growth through 15-keto-PGE2/PPAR?-mediated activation of p21WAF1/Cip1 Oncogene doi:10.1038/onc.2013.69
[PTGR2 ]
A Modified Form of Diphthamide Causes Immunotoxin Resistance in a Lymphoma Cell Line with a Deletion of the WDR85 Gene J. Biol. Chem., Apr 2013; 288: 12305 - 12312.
[WDR85]
A New Signaling Pathway (JAK-Fes-phospholipase D) That Is Enhanced in Highly Proliferative Breast Cancer Cells J. Biol. Chem., Apr 2013; 288: 9881 - 9891.
[FES]
Androgen receptor inclusions acquire GRP78/BiP to ameliorate androgen-induced protein misfolding stress in embryonic stem cells Cell Death and Disease 4, e607 doi:10.1038/cddis.2013.122
[GRP78]
KCNA5 Other names: ATFB7; HCK1; HK2; HPCN1; KV1.5; PCN1
Host Cell
CHL
cDNA Clone:
SC123964
Format:
2 x 1 mL frozen cell vials each containing 1 x 10E6 cells
Mycoplasma:
Negative by DNA staining and direct culture methods (ATTC - detailed information available upon request)
Cell Line Validation:
Gene expression: qPCR experiments determined specific over-expression of KNA5A.Figure 1.
Functional validation
Current-voltage relationship. Figure 2
Expression statistics. Figure 3
Pharmacology (Figure 4) - Inhibition of hKv1.5 K+ currents by known K+ channel blockers such as Psora-4, CP-339818, DPO-1, Mephetyl Tetrazole, Bupivicaine, Quinidine
Background:
Kv1.5 is a voltage-gated K+ channel which underlies the ultra-rapidly activating delayed rectifier K+ current (Ikur) found in human atrial myocytes. Kv1.5 is also expressed in the human ventricle where it is possible it contributes to the K+ current through formation of heteromultimeric K+ channels with other Kv-alpha subunits. Inhibition of Kv channels increases atrial refractory period in man – this is clinically valuable for the treatment of patients suffering atrial fibrillation, but undesirable for drugs designed to have inert cardiovascular profiles.
Figure 1: In a SYBR green qPCR experiment, specific over-expression of KCN5A was determined using gene specific primers. Data are shown as fold over-expression after normalization against GAPDH.
Figure 2: Current-voltage relationship for hKv1.5 channels stably expressed in CHL cells (A). Representative recordings from CHL-Kv1.5 (B) and CHL-wild type cells (C). Experiments were conducted from a holding potential of -80mV. The depolarising step length was 200ms.
Figure 3: Expression profile: outward K+ currents of >0.6 nA were observed in 305 of 310 cells (98%), with a mean amplitude of 2.03 ± 0.85 nA (n=310; mean ± S.D.). The peak current in each cell, evoked from a depolarising pulse to +40 mV, was divided into 0.2 nA bins to create the population histogram.
Figure 4: Summary pharmacology of hKv1.5-CHL cell line. Voltage-clamp recordings were made using repeated gating steps, Vh -80mV, Vstep +40mV, 1 Hz, pulse duration 250ms. Control traces in black, test compounds in red. IC50 values were obtained by fitting concentration-response curves to values for inhibition of the 1st and 5th pulse in the train (µM, shown in table). Use-dependent and open channel block can be assessed within this protocol (note blue arrow).
