Abstract
BackgroundThe metastatic ability of tumor cells is determined by level of expression of specific genes that may be identified with the aid of cDNA microarray containing thousands of genes and can be used to establish the expression profile of disease related genes in complex biological system.Materials and MethodsSalivary adenoid cystic carcinoma cell line and its high metastases adenoid cystic carcinoma clone were used as model systems to reveal the gene expression alteration related to metastasis mechanism by cDNA microarray analysis. The correlation of metastatic phenotypic changes and expression levels of 4 selected genes (encoding CD98, L6, RPL29, and TSH) were further validated by using RT-PCR analysis of human tumor specimens from primary adenoid cystic carcinoma and corresponding metastasis lymph nodes.ResultsOf the 7,675 clones of known genes and expressed sequence tags (ESTs) that were analyzed, 30 showed significantly different (minimum 3 fold) expression levels in two cell lines. Out of 30 genes found differentially expressed, 18 were up regulated (with ratio more than 3) and 12 down regulated (with ratio less than 1/3).ConclusionSome of these genes are known to be involved in human tumor antigen, immune surveillance, adhesion, cell signaling pathway and growth control. It is suggested that the microarray in combination with a relevant analysis facilitates rapid and simultaneous identification of multiple genes of interests and in this study it provided a profound clue to screen candidate targets for early diagnosis and intervention.
Highlights
The metastatic ability of tumor cells is determined by level of expression of specific genes that may be identified with the aid of cDNA microarray containing thousands of genes and can be used to establish the expression profile of disease related genes in complex biological system
Semi-quantitative RT-PCR Four selected genes, CD98, L6, RPL29, and thyroid-stimulating hormone (TSH), were selected and investigated in this study to evaluate the difference of expression of them between primary Adenoid cystic carcinoma (ACC) tumor tissue and ACC metastasis lymph node in vivo, confirming the reliability of our approach. 1 μg each of total RNA from 4 primary tumor tissue samples and 4 corresponding metastasis lymph node tissue samples were used as templates in a total volume of 20 μl reverse transcription reaction system
Identification of genes differentially expressed between ACC-2 and ACC-M cells The cDNA microarray technique was applied to analyze the expression patterns of ACC-2 and ACC-M cells
Summary
Cell lines and specimens Cell lines ACC-2 and clone ACC-M were previously established in the tumor biology laboratory of Shanghai Ninth Hospital in Shanghai Second Medical University. One specimen was from primary tumor tissue and the other from metastatic lymph node. Both of them were confirmed by histological examination. Total RNA was treated with DNase (Boehringer Mannheim, Germany) and RNasin (Promega, Madison, USA) at 37°C for 15 minutes to remove contaminated DNA. Semi-quantitative RT-PCR Four selected genes, CD98, L6, RPL29, and TSH, were selected and investigated in this study to evaluate the difference of expression of them between primary ACC tumor tissue and ACC metastasis lymph node in vivo, confirming the reliability of our approach. 1 μg each of total RNA from 4 primary tumor tissue samples and 4 corresponding metastasis lymph node tissue samples were used as templates in a total volume of 20 μl reverse transcription reaction system. Amplified fragments were visualized by ethidium bromide staining of the agarose gel and photographed under UV light
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
