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Archive > Volume 21 Issue 2 > 2014,21(2):175-180. DOI:10.3872/j.issn.1007-385X.2014.02.011 Prev Next
Clinical Research
Expression aberration and methylation of SRY-Box2 gene in gastric tumor tissues
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Abstract:
Objective : To determine mRNA levels and methylation status of stem cell transcription factor SRY-Box2 in gastric cancer (GC) cell lines (MKN74, MKN45) and tissue specimens in association with pathogenesis and clinicopathological features of GC. Methods: Eighty-six gastric cancer patients diagnosed in the Thoracic Surgery of the Fourth Hospital of Hebei Medical University between 2007 and 2012 were recruited. Biopsy specimens were collected from primary tumors and the corresponding adjacent tissues. The gastric cancer cell lines (MKN74, MKN45) were treated respectively with 5-aza-2’-deoxycytidine (5-Aza-Dc) and trichostatin A (TSA). Levels of CpG methylation of the SRY-Box2 promoter and SRY-Box2 mRNA were assessed by methylation specific PCR (MSP) and RT-PCR, respectively, in MKN74, MKN45 cells after drug treatments and in biopsy specimens. The relevance of SRY-Box2 gene methylation to and clinicopathological features of the cancer and to changes in RY-Box2 mRNA abundance was analyzed. Results: SRY-Box2 mRNA was detected in MKN45 cells but not in MKN74 cells. Treatment with 5-aza-2’-deoxycytidine (5-Aza-Dc, a demethylation agent) significantly increased SRY-Box2 mRNA abundance but trichostatin A (TSA) had no effect inMKN45 cells. Hypermethylation of SRY-Box2 gene containing a CpG island was observed in MKN74 cells. The frequency of expression loss of the SRY-Box2 gene (19.8% vs 7.0%; χ 2=69.073, P=0.014) and the level of hypermethylation (14.0% vs 1.2%；χ 2=10.069, P=0.002) were all significantly higher in the cancer tissue as compared with adjacent non-cancerous tissues.and hypermethylation was significantly correlated with expression loss in the SRY-Box2 gene (χ 2=50.878,P<0001). Furthermore, SRY-Box2 gene hypermethylation status was also correlated with lymph node metastasis (χ 2=3947, P=0.047), but not with clinical stage, pathological grade and depth of invasion ( P>0.05). Conclusion: CpG hypermethylation may be one of the mechanisms responsible for the expression loss of the SRY-Box2 gene and may play some role in the pathogenesis of gastric cancer.
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Project supported by the Natural Science Foundation of Hebei Province（No. H2013206315）
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