Construction of transcriptome profiling of cancerous parenchyma cells using laser capture microdissection and Gene-Chip technology
10.3781/j.issn.1000-7431.2008.09.020
- Author:
Yu-Hao SUN
1
Author Information
1. Department of General Surgery
- Publication Type:Journal Article
- Keywords:
Colonic neoplasms;
Lasers;
Microdissection;
Molecular probe techniques
- From:
Tumor
2008;28(9):802-804
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct transcriptome profiling of parenchyma cell using laser capture microdissection (LCM) combined with Gene-Chip technology. Methods: We obtained cancerous parenchyma cells from frozen sections of tumor tissues by using LCM. RNA was extracted from the cells. aRNA was obtained after linear amplification, and then converted to cRNA probe, finally hybridized with Affymetrix HG-U133. Plus 2.0 Gene-Chip for construction of full genome transcriptome profiling. Results: About 3 mm2 cancerous colon cells were obtained from each of 5 colon cancer specimens, respectively. The quantity of total RNA extracted from cancerous colon cells ranged from 118.4 to 300.3 ng. We obtained 7 υg of aRNA after linear amplification from 100 ng mixed RNA. The quality control was guided by the operating instruction standard of Gene-Chip. Totally 18 205 transcripts were presented representing 15 276 genes. Conclusion: The combination of laser capture microdissection and Gene-Chip technology could be used for construction of transcriptome profiling of purified cancerous parenchyma cells.
