0 software and a P value Poziotinib mw < 0.05 was considered statistically significant. Results RT-PCR and real time RT-PCR analysis The expression levels of lamin A/C mRNA were examined in 52 paired clinical samples by semiquantitative RT-PCR. As shown in Fig. 1A, lamin A/C mRNA could be detected in GC tissues as well as in matched

non-cancerous tissues. However, a large decrease in the levels of lamin A/C mRNA expression was observed in primary GC as compared with normal tissue. The analysis of results displayed the density value (normalized to β-actin expression as a loading control) of tumour was significantly lower than that in AZD3965 mw corresponding non-cancerous tissue using paired t-test (p = 0.011, Fig. 1B). Figure 1 Expression pattern of lamin A/C in GC specimens

by RT-PCR. (A) 1.5% agarose electrophoresis of lamin A/C products of RT-PCR in GC specimens. Representative results from 4 pairs of GC and corresponding normal gastric tissues are shown. β-actin was used as an internal quantitative control. (B) Densitometry analyses of lamin A/C mRNA level quantified by compared with β-actin in GC and corresponding normal gastric samples. The expression of lamin A/C gene was reduced in tumour tissues when compared with corresponding non-tumourous tissues (p = 0.011). T, GC; N, corresponding non-cancerous tissues. To validate the results BVD-523 in vitro of semiquantitative RT-PCR, we randomly selected 30 cases out of the 52 patients to investigate the mRNA expression level with real time RT-PCR. The dissociation Phosphoprotein phosphatase curve and amplification curve were shown in Fig. 2A and 2B. The fold change in expression levels determined by a comparative

CT method also demonstrated that lamin A/C expression is reduced in GC tissues. We further analyzed the correlations between lamin A/C mRNA expression and clinicopathological features. As shown in Table 1, the mRNA expression level was evidently lower in poor differentiated tumours than that in well or moderately differentiated tumours. Decreased of lamin A/C expression correlated with histological differentiation significantly (r = 0.438, p = 0.025). However, there were no statistical correlations between lamin A/C and invasion, tumour size and metastasis. Table 1 Correlations between lamin A/C expression detected by real time RT-PCR and pathological variables in 30 cases of GC Variables Number of Cases Fold Change (mean ± SD) t p -Value Invasion            Profound layer 24 0.77 ± 0.19 -0.692 0.495    Superficial layer 6 0.83 ± 0.19     Differentiation            Poor 21 0.73 ± 0.19 -2.376 0.025a    Well or Moderate 9 0.90 ± 0.13     Metastasis            No 23 0.76 ± 0.18 -0.792 0.435    Yes 7 0.83 ± 0.23     Tumour Size (cm)            < 5 18 0.83 ± 0.18 1.704 0.099    ≥5 12 0.71 ± 0.20     a Statistically significant (p < 0.05). Figure 2 The dissociation curves and amplification curves of lamin A/C in GC specimens by real time RT-PCR.