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. 2006 Nov 14;12(42):6893-7.
doi: 10.3748/wjg.v12.i42.6893.

Identification of differentially expressed genes in mouse hepatocarcinoma ascites cell line with low potential of lymphogenous metastasis

Xiao-Nan Cui  1 , Jian-Wu Tang, Li Hou, Bo Song, Li-Ying Ban
Affiliations

Identification of differentially expressed genes in mouse hepatocarcinoma ascites cell line with low potential of lymphogenous metastasis

Xiao-Nan Cui et al. World J Gastroenterol. .

Abstract

Aim: To identify genes differentially expressed in mouse hepatocarcinoma ascites cell line with low potential of lymphogenous metastasis.

Methods: A subtracted cDNA library of mouse hepatocarcinoma cell line with low potential of lympho-genous metastasis Hca-P and its synogenetic cell line Hca-F with high metastatic potential was constructed by suppression subtracted hybridization (SSH) method. The screened clones of the subtracted library were sequenced and GenBank homology search was performed.

Results: Fifteen differentially expressed cDNA fragments of Hca-P were obtained which revealed 8 known genes, 4 expressed sequence tags (ESTs) and 3 cDNAs showed no homology.

Conclusion: Tumor metastasis is an incident involving multiple genes. SSH is a useful technique to detect differentially expressed genes and an effective method to clone novel genes.

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Figures

Figure 1
Figure 1
The effect of RsaIdigestion. Lane 1, 3: cDNA of Hca-F and Hca-P cells; Lane 2, 4: cDNA of Hca-F and Hca-P cells after RsaIdigestion; M: DNA marker DL2000.
Figure 2
Figure 2
Ligation efficiency analysis. Lane 1: Tester-1 as template, G3PDH 3’Primer and PCR Primer1; Lane 2: Tester-1 as template, G3PDH 3‘Primer and G3PDH 5‘Primer; Lane 3: Tester-2R as template, G3PDH 3‘Primer and PCR Primer1; Lane 4: Tester-2R as template, G3PDH 3‘Primer and G3PDH 5‘Primer.
Figure 3
Figure 3
The results of secondary PCR amplification. Lane 1-3: Product of primary PCR amplification, Lane 4: secondary PCR amplification product of unsubtracted cDNA, Lane 5: secondary PCR amplification product of subtracted cDNA, Lane 6: secondary PCR amplification product of PCR control cDNA, M: DNA Marker DL2000.
Figure 4
Figure 4
Analysis of subtraction effect. PCR was performed on subtracted (Lane 1-4) or unsubtracted (Lane 5-8) secondary PCR product with G3PDH 5’Primer and 3’primer. Lanes 1, 5: 20 cycles, Lanes 2, 6: 25 cycles, Lanes 3, 7: 30 cycles, Lanes 4, 8: 35 cycles. M: DNA marker DL2000.
Figure 5
Figure 5
The results of clone PCR amplification. There was an average insert size of 300-1000 bp. M: DNA marker DL2000.
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