Quantitative analysis of tumor mitochondrial RNA using microarray.
OBJECTIVE
To design a novel method to rapidly detect the quantitative alteration of mtRNA in patients with tumors.


METHODS
Oligo 6.22 and Primer Premier 5.0 bio-soft were used to design 15 pairs of primers of mtRNA cDNA probes in light of the functional and structural property of mtDNA, and then RT-PCR amplification was used to produce 15 probes of mtRNA from one normal gastric mucosal tissue. Total RNA extracted from 9 gastric cancers and corresponding normal gastric mucosal tissues was reverse transcribed into cDNA labeled with fluorescein. The spotted mtDNA microarrays were made and hybridized. Finally, the microarrays were scanned with a GeneTAC laser scanner to get the hybridized results. Northern blot was used to confirm the microarray results.


RESULTS
The hybridized spots were distinct with clear and consistent backgrounds. After data was standardized according to the housekeeping genes, the results showed that the expression levels of some mitochondrial genes in gastric carcinoma were different from those in the corresponding non-cancerous regions.


CONCLUSIONS
The mtDNA expression microarray can rapidly, massively and exactly detect the quantity of mtRNA in tissues and cells. In addition, the whole expressive information of mtRNA from a tumor patient on just one slide can be obtained using this method, providing an effective method to investigate the relationship between mtDNA expression and tumorigenesis.
Han
Cheng-Bo
CB
Cancer Institute, First Affiliated Hospital, China Medical University, Shenyang 110001, Liaoning Province, China. hancb@126.com
Mao
Xiao-Yun
XY
Xin
Yan
Y
Wang
Shao-Cheng
SC
Ma
Jia-Ming
JM
Zhao
Yu-Jie
YJ
D015152:Blotting, Northern; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D046228:Microarray Analysis / Q000379:methods*; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000032:analysis / Q000235:genetics*; D000077278:RNA, Mitochondrial; D012334:RNA, Neoplasm / Q000032:analysis / Q000235:genetics*; D013274:Stomach Neoplasms / Q000235:genetics*
D000077278:RNA, Mitochondrial; D012334:RNA, Neoplasm; D012313:RNA