β-连环蛋白在肝癌细胞核内的积累上调谷胱甘肽 S-转移酶 M3 mRNA。
β-catenin accumulation in nuclei of hepatocellular carcinoma cells up-regulates glutathione-s-transferase M3 mRNA.
机构信息
Department of Pharmacology, College of Pharmacy, South-Central University for Nationalities, No. 708, Minyuan Road, Wuhan 430074, Hubei Province, China.
出版信息
World J Gastroenterol. 2011 Apr 7;17(13):1772-8. doi: 10.3748/wjg.v17.i13.1772.
AIM
To identify the differentially over-expressed genes associated with β-catenin accumulation in nuclei of hepatocellular carcinoma (HCC) cells.
METHODS
Differentially expressed genes were identified in radiation-induced B6C3 F1 mouse HCC cells by mRNA differential display, Northern blot and RT-PCR, respectively. Total glutathione-s-transferase (GST) activity was measured by GST activity assay and β-catenin localization was detected with immunostaining in radiation-induced mouse HCC cells and in HepG2 cell lines.
RESULTS
Two up-regulated genes, glutamine synthetase and glutathione-s-transferase M3 (GSTM3), were identified in radiation-induced mouse HCC cells. Influence of β-catenin accumulation in nuclei of HCC cells on up-regulation of GSTM3 mRNA was investigated. The nearby upstream domain of GSTM3 contained the β-catenin/Tcf-Lef consensus binding site sequences [5'-(A/T)(A/T) CAAAG-3'], and the total GST activity ratio was considerably higher in B6C3F1 mouse HCC cells with β-catenin accumulation in nuclei of HCC cells than in those without β-catenin accumulation (0.353 ± 0.117 vs. 0.071 ± 0.064, P < 0.001). The TWS119 (a distinct GSK-3β inhibitor)-induced total GST activity was significantly higher in HepG2 cells with β-catenin accumulation than in those without β-catenin accumulation in nuclei of HCC cells. Additionally, the GSTM3 mRNA level was significantly higher at 24 h than at 12 h in TWS119-treated HepG2 cells.
CONCLUSION
β-catenin accumulation increases GST activity in nuclei of HCC cells, and GSTM3 may be a novel target gene of the β-catenin/Tcf-Lef complex.
目的
鉴定与肝癌（HCC）细胞中β-catenin 核内积累相关的差异表达基因。
方法
通过 mRNA 差异显示、Northern blot 和 RT-PCR，分别在辐射诱导的 B6C3F1 小鼠 HCC 细胞中鉴定差异表达基因。通过 GST 活性测定和免疫染色检测辐射诱导的小鼠 HCC 细胞和 HepG2 细胞系中的总谷胱甘肽-S-转移酶（GST）活性和β-catenin 定位。
结果
在辐射诱导的小鼠 HCC 细胞中鉴定出两个上调基因，即谷氨酰胺合成酶和谷胱甘肽-S-转移酶 M3（GSTM3）。研究了 HCC 细胞中β-catenin 核内积累对 GSTM3 mRNA 上调的影响。GSTM3 的近端上游区域包含β-catenin/Tcf-Lef 共识结合位点序列[5'-(A/T)(A/T) CAAAG-3']，并且在具有β-catenin 核内积累的 B6C3F1 小鼠 HCC 细胞中，总 GST 活性比没有β-catenin 积累的细胞（0.353 ± 0.117 比 0.071 ± 0.064，P < 0.001）显著更高。在具有β-catenin 核内积累的 HepG2 细胞中，TWS119（一种独特的 GSK-3β 抑制剂）诱导的总 GST 活性明显高于没有β-catenin 积累的细胞。此外，在 TWS119 处理的 HepG2 细胞中，GSTM3 mRNA 水平在 24 小时时明显高于 12 小时。
结论
β-catenin 核内积累增加了 HCC 细胞中的 GST 活性，GSTM3 可能是β-catenin/Tcf-Lef 复合物的一个新的靶基因。
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