The rapid activation of the Lgls1 gene (Figure 5) indicates that VPA may act at the immediate level of the Lgals1 promoter.
By comparing gene expression changes in whole embryos with those in a cultured cell model, we defined a subset of VPA-responsive genes that may be evaluated as potential biomarkers of VPA teratogenicity (Lgals1, Idl, fasn, Anxa5).
Forward primer Gene symbol Forward primer sequence concentration (nM) Kpnb1 5'-GGGAATCGTCCAGGGATTG- 3' 900 Krtl-18 5'-AATCGAGGCACTCAAGGAAGAA- 3' 300 Lgals1 5'-GAATCTCTTCGCTTCAGCTTCA- 3' 50 Mt2 5'-CGCCATGGACCCAACT- 3' 50 Upp 5'-TCACCATCATCCGCATTGG- 3' 300 Vcl 5'-TGCCAAGCAGTGCACAGATAA- 3' 50 Ppia 5'-TTCCTCCTTTCACAGAATTATTCCA- 3' 50 Reverse primer Gene symbol Reverse primer sequence concentration (nM) Kpnb1 5'-AAATAAATTCTACTCTGGCTGTACCA- 3' 900 Krtl-18 5'-GGCATCCACTTCCACGTCA- 3' 300 Lgals1 5'-CAGGTTTGAGATTCAGGTTGCT- 3' 50 Mt2 5'-AGGAAGTACATTTGCATTGTTTGC- 3' 50 Upp 5'-GCCTGCTGCGTGATGACA- 3' 900 Vcl 5'-GGTCCGGCCCAGCATAGT- 3' 50 Ppia 5'-CCGCCAGTGCCATTATGG- 3' 50 Amplicon Gene symbol length (nt) Kpnb1 83 Krtl-18 112 Lgals1 68 Mt2 89 Upp 73 Vcl 124 Ppia 75 Table 2.
 observed increased expression of the LGALS1 mRNA in the stomach cell line TMC-1, suggesting that this protein is important in the metastasis process.
When we analyzed the association of ANXA1 and LGALS1 mRNA expression levels with the risk factors in the two lesions studied, we only found an association between the relative expression of LGALS1 and gender in the IM group, with women showing higher expression than men (RQ = 0.80 versus -0.15).
Other studies suggest that sex steroids may regulate LGALS1 expression in female reproductive tissues of humans and rats [70, 71].
Table 2: Relative gene expression of ANXA1 and LGALS1 mRNA in the intestinal metaplasia (IM) and gastric ulcer (GU) groups and comparison according to risk factors.
In another analysis, we investigated the GA group for a possible association between ANXA1 and LGALS1 mRNA expression and risk factors such drinking, smoking, and histological type of gastric cancer (Table 4), but no association was observed.
For LGALS1, our study showed a slightly increased relative expression only in 60% of the GA cases (mean RQ = 2.44), with an increase of 2 to 7 times, but in chronic gastritis the mean value was low (mean RQ = 0.43).
In contrary, the LGALS1 mRNA levels were lower in the CG group in comparison with the GA group.
The comparison between the CG and GA groups regarding ANXA1 and LGALS1 mRNA levels and risk factors did not show any association with the LGALS1 mRNA levels.