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. 2013 Feb;25(1):71-8.
doi: 10.3978/j.issn.1000-9604.2012.12.03.

Primarily screening and analyzing ESTs differentially expressed in rats' primary liver cancer

Dandan Liu  1 , Lijuan Zhi, Mingxia Ma, Dan Qiao, Meijuan Wang, Yawei Wang, Baijie Jin, Anqi Li, Guting Liu, Yiqing Zhang, Yanyan Song, Hongxu Zhang
Affiliations

Primarily screening and analyzing ESTs differentially expressed in rats' primary liver cancer

Dandan Liu et al. Chin J Cancer Res. 2013 Feb.

Abstract

Objective: To screen and analyze key express sequence tags (ESTs) which were differentially displayed in every period of SD rats' primary hepatic carcinoma and reveal the molecular mechanism of carcinogenesis.

Methods: Using diethylnitrosamine (DENA) as a cancerigenic agent, animal models with different phases of primary hepatic cancer were constructed in SD rats. Rats were respectively sacrificed at d 14, d 28, d 56, d 77, d 105 and d 112 after the rats received DENA by gavage, then the livers were harvested. One part of the livers was classified according to their pathological changes, while the other was reserved for molecular mechanism studies on hepatocarcinogenesis. The differentially expressed genes were isolated from both normal and morbid tissues by mRNA differential display technique (DDRT-PCR). After the fragments were sequenced, bioinformatics were used to analyze the results.

Results: Twelve differentially expressed cDNA fragments were obtained. Nine fragments had the homology with known cDNA clones, especially EST-7 was similar to BN/SsNHsdMCW mitochondrion gene and the identity was 100% which suggested EST-7 may be the part of BN/SsNHsdMCW mitochondrion gene. In contrast, other three fragments (EST-1, EST-3 and EST-5) had extremely low identity to any genes registered in GENBANK databases.

Conclusions: BN/SsNHsdMCW mitochondrion gene was expressed in different periods of hepatocarcinogenesis. Moreover, EST-1, EST-3 and EST-5 were suggested to contribute to the development of rat hepatocarcinogenesis, and thus may be candidates of new targets of oncogenes or cancer suppressor genes.

Keywords: Animal models of primary liver cancer; DDRT-PCR (differential display reverse transcription PCR); ESTs (express sequence tags); mitochondrion gene.

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Figures

Figure 1
Figure 1
Results of RNA isolation
Figure 2
Figure 2
Results of cDNA identification. M. molecular weight standard; C. the negative control amplified with RNA as the template
Figure 3
Figure 3
Differential expressed genes among differential pathological stages. Parts of the differentially expressed bands were showed by arrows. N. normal liver; 1. light inflammation; 2. heavy inflammation; 3. nodus hyperplasia; 4. untypical nodus hyperplasia; 5. intrahepatic cholangiocarcinoma; M. molecular weight standard
Figure 4
Figure 4
Parts of the interesting DNA bands were reamplified and showed by agarose gels. M, molecular weight standard; C, the negative control amplified with H2O as the template
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