期刊
ELECTROPHORESIS
卷 31, 期 22, 页码 3763-3770出版社
WILEY
DOI: 10.1002/elps.201000265
关键词
Chemoresistance; Lung cancer; Microfluidic device; SPCA1; VP-16
资金
- National Natural Science Foundation of China [30470464, 30670532]
Microchip-based systems have been developed rapidly due to their desirable advantages over conventional platforms. Higher level system integration and complex microdevices are emerging to satisfy the demand for high-throughput and large-scale applications. However, most of the devices need to be fabricated with complicated microvalves and micropumps, which, to some extent, limit the use of the novel technique. In this study, a simple microdevice was developed to perform chemotherapy resistance analysis in lung cancer cell line SPCA1. This device includes a PDMS chip for which a simple external small dip served as a microvalve to control the fluid flow so that the parallel control experiment could be carried out simultaneously, and a syringe pump, which supplied the cells with fresh medium mimicking the microenvironment in vivo. Cell culture, detection of drug resistance related protein P-glycoprotein (P-gp) and glutathione S-transferase-pi (GST-pi) and cell viability after VP-16 treatment on experimental (pretreated with corresponding inhibitors) and control groups were achieved. The results demonstrated that the cells could grow and spread well for at least 3 days. The expression of P-gp and GST-pi was obviously downregulated by corresponding inhibitors. The percentage of apoptotic cells for P-gp inhibition group increased 2.9-fold compared with that of control group (23.7 +/- 2.6 versus 8.1 +/- 3.0%, p <0.05), while for GST-pi inhibition, there was no obvious distinction between the experimental and control group. The simple microdevice is capable of integrating parallel operations involving cell culture and functional analysis, offering an easy and flexible platform for a stable long-term cell culture and comparison research.
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