Electrotaxis Study of Human Lung Cancer Cells by Using Microfluidic Chips
Date Issued
2012
Date
2012
Author(s)
Huang, Ching-Wen
Abstract
Electrotaxis is the movement of adherent living cells in response to a direct current (dc) electric field (EF) of physiological strength. In this work, microfluidic cell culture chips were developed to be used for long-term electrotaxis study on a microscope. The cellular response under three different electric field strengths was studied in a single channel microfluidic chip. EF inside the micro-chamber was numerically simulated and compared to the measured value. Lung cancer cell lines with high and weak metastasis potential, CL1-5 and CL1-0, respectively, were used to demonstrate the function of the multi-field chip (MFC). The two cell lines exhibited greatly different response under the applied EF of E = 74-375mV/mm. CL1-5 cells migrated toward the anode while CL1-0 cells did not show obvious response. Under the applied EF, cell orientation was observed accompanying the cell migration in CL1-5.
To understand the transcriptional response of CL1-5 and CL1-0 cells to a dcEF, microarray analysis was performed in this study. Affymetrix GeneChip was used for microarray analysis. A large electric-field chip (LEFC) was designed, fabricated, and used for sample collection. CL cells were treated with the EF strength of 0mV/mm (the control group) and 300mV/mm (the EF-treated group) for two hours. Signaling pathways involving the genes that expressed differently between the two groups were revealed. In CL1-5, it was shown that the EF-regulated genes highly correlated to adherens junction, telomerase RNA component gene regulation, and tight junction. In CL1-0, the same analysis revealed that the EF-regulated genes were mainly involved in translation, purine metabolism, and cell apoptosis. In other words, the EF-regulated genes in CL1-0 did not show strong correlation to cell adhesion and migration. We further observed a high percentage of significantly regulated genes which encode cell membrane proteins in CL1-5, suggesting that dcEF may directly influence the activity of cell membrane proteins in signal transduction. However, the dcEF did not have significant influence in membrane protein encoding genes in CL1-0.
Our study demonstrated that CL1-0 and CL1-5 not only showed distinct activities induced by the dcEF, they also showed different gene expression changes. Some of the EF-regulated genes have been reported to be essential whereas others are novel for electrotaxis. Our result confirms that the regulation of gene expression is involved in the mechanism of electrotactic response. By building an interaction network of EF-regulated genes and considering the original gene expression differences of the two cell lines, we proposed several membrane proteins that may sense the EF signal and cause the different electrotactic responses between the high and low metastatic cancer cells.
Subjects
Electrotaxis
Microfluidic
Metastasis
Lung cancer cell
Gene expression
Microarray analysis
SDGs
Type
thesis
File(s)![Thumbnail Image]()
Loading...
Name
ntu-101-D95548007-1.pdf
Size
23.54 KB
Format
Adobe PDF
Checksum
(MD5):97357181d502c03b74888260efce95c9