Method of measuring cell-substrate impedance in living cells to identify compounds affecting receptor tyrosine kinase (RTK) activity and pathways
A method of identifying a potential therapeutic compound that affects a Receptor Tyrosine Kinase (RTK) pathway in cancer cells, which includes: providing a device capable of measuring cell-substrate impedance; culturing cancer cells in serum-free media in at least two wells of the device; adding to a first well a proposed therapeutic compound that affects a RTK pathway and a RTK stimulating factor for the RTK pathway to form a test well, and adding to another well the RTK stimulating factor to form a control well; continuously monitoring cell-substrate impedance of the at least two wells and optionally determining cell indices from the monitored cell-substrate impedance; and determining a difference in impedance or optionally cell index between the test well and control well; and if significantly different, concluding the proposed therapeutic compound is therapeutically active in the RTK pathway within the cancer cells.
1. A method of identifying a potential therapeutic compound that affects a Receptor Tyrosine Kinase (RTK) pathway in cancer cells, the method comprising:
a) providing a device capable of measuring cell-substrate impedance, wherein the device comprises at least two wells, each well comprising an electrode array, further wherein the device is operably connected to an impedance analyzer;
b) culturing cells embodied as cancer cells in serum-free media in the at least two wells;
c) adding to a first well a proposed therapeutic compound that affects an RTK pathway and an RTK stimulating factor for the RTK pathway to form a test well, and adding to another well the RTK stimulating factor to form a control well;
d) continuously monitoring cell-substrate impedance of the at least two wells and optionally determining cell indices from the monitored cell-substrate impedance; and
e) determining a difference in impedance or optionally in cell index between the test well and control well; and if significantly different,
f) concluding the proposed therapeutic compound is therapeutically active in the RTK pathway within the cancer cells.
2. The method of claim 1 , wherein the cancer cells are primary cells.
3. The method according to claim 1 , wherein the cancer cells are selected from the group consisting of breast cancer cells, lung cancer cells, and colon cancer cells.
4. The method according to claim 1 , wherein the cancer cells are selected from the group consisting of ovarian cancer cells, prostate cancer cells, hepatosarcoma cells, epidermoid cancer cells, and fibrosarcoma cells.
5. The method of claim 1 , wherein the proposed therapeutic compound is a kinase inhibitor.
6. The method of claim 1 , wherein the proposed therapeutic compound is Trastuzumab (HERCEPTIN) or 4- Quinazolinamine N-(3-chloro-4-fluorophenyl)-7-methoxy-6-[3-(4-morpholinyl) propoxy](GEFITINIB).
7. The method of claim 1 , wherein the proposed therapeutic compound is added before the RTK stimulating factor.
8. The method of claim 1 , wherein the RTK stimulating factor is a growth factor.
9. The method of claim 1 , wherein the RTK stimulating factor is selected from the group consisting of epidermal growth factor (EGF), platelet derived growth factor (PDGF), a nerve growth factor (NGF), hepatocyte growth factor (HGF), fibroblast growth factor (FGF), and an antibody capable of activating RTK.