Frequent downregulation of the runt domain transcription factors RUNX1, RUNX3 and their cofactor CBFB in gastric cancer
Frequent downregulation of the runt domain transcription factors RUNX1, RUNX3 and their cofactor CBFB in gastric cancer
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DOI：
10.1002/ijc.20551
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发表时间：
2005-01
影响因子：
6.4
通讯作者：
C. Sakakura;A. Hagiwara;Koji Miyagawa;S. Nakashima;T. Yoshikawa;S. Kin;Y. Nakase;Kosei Ito;H. Yamagishi;S. Yazumi;T. Chiba;Yoshiaki Ito
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作者：
C. Sakakura;A. Hagiwara;Koji Miyagawa;S. Nakashima;T. Yoshikawa;S. Kin;Y. Nakase;Kosei Ito;H. Yamagishi;S. Yazumi;T. Chiba;Yoshiaki Ito
我们以前的研究表明，RUNX3功能缺失与人类胃癌的发生发展有因果关系，但RUNX家族其他成员的潜在作用尚未见报道。我们检测了RUNX1、RUNX2和Cbfb 3个RUNX基因在胃癌细胞系和原发胃癌标本中的表达，并与早期报道的RUNX3结合临床病理因素进行了比较。采用Northern印迹分析、定量RT-PCR和原位杂交方法检测了9株胃癌细胞系、56例原发胃癌组织和癌旁正常胃粘膜组织中RUNX家族基因的表达。Northern印迹分析显示RUNX1和RUNX3分别在67%和78%的受试细胞系中表达下调。RUNX1、RUNX2、RUNX3、Cbfb的平均RUNX m RNA/β-Actin m RNA比值(×10 3)分别为48.0±2 1.1vs.2 1.4±8.1、1.1±0.3、9.2±6.3、3.1±1.3和42.0±19.4、2 1.0±8.4(平均标准差)。RUNX2的基础表达很弱，在胃癌组织中无明显变化。定量RT-PCR分析显示，RUNX1和RUNX3分别在62%和69%的手术切除标本中与周围粘膜相比显著下调(p<；0.01)。此外，编码RUNX1、-2、-3辅助因子的基因Cbfb也显著下调(32%，p<；0.05)。RUNX1、RUNX3和Cbfb基因下调的百分比随着肿瘤分期的进展而增加。Tricostatin A和5‘-azacitidin使RUNX3表达恢复，但不能恢复RUNX1和Cbfb在胃癌细胞中的表达，提示RUNX1和Cbfb的下调是由于启动子区域甲基化以外的机制。这些结果表明，除RUNX3外，RUNX1和Cbfb在胃癌中也有一定的作用，RUNX基因家族在胃癌中的作用比以往认识的更广泛和复杂。
Our previous studies suggest that lack of RUNX3 function is causally related to the genesis and progression of human gastric cancer, but potential roles of other members of the RUNX family genes have not yet been reported. We examined the expression of 3 Runt‐related (RUNX) genes, RUNX1, RUNX2 and CBFB, in gastric cancer cell lines and primary gastric cancer specimens and compared them to those of RUNX3 reported earlier in conjunction with clinicopathologic factors. Expression of RUNX family genes in 9 gastric cancer cell lines, 56 primary gastric cancer specimens and surrounding normal gastric mucosa were estimated by Northern blot analysis, quantitative RT‐PCR and in situ hybridization. Northern blot analysis in gastric cancer cell lines showed downregulation of RUNX1 and RUNX3 in 67% and 78% of the cell lines tested, respectively. The ratio of the average RUNX mRNA/β‐actin mRNA ratio (×103) for RUNX1 was 48.0 ± 21.1 vs. 21.4 ± 8.1; RUNX2, 1.1 ± 0.3 vs. 1.0 ± 0.2; RUNX3, 9.2 ± 6.3 vs. 3.1 ± 1.3 and CBFB, 42.0 ± 19.4 vs. 21.0 ± 8.4 (normal vs. tumor, respectively, average ±SD). The basal RUNX2 expression was very weak, and there was no significant change in gastric cancers. Both RUNX1 and RUNX3 showed remarkable downregulation in 62% and 69%, respectively, of surgically resected specimens compared to surrounding mucosa analyzed by quantitative RT‐PCR (p < 0.01). Furthermore, CBFB, the gene encoding the cofactor of RUNX1, ‐2, ‐3, was also downregulated in significant fraction (32%, p < 0.05). The percentage of downregulation of RUNX1, RUNX3 and CBFB increased as the cancer stage progressed. Tricostatin A and 5′‐azacitidin reactivate RUNX3 expression, but they could not reactivate expression of RUNX1 and CBFB in gastric cancer cells, suggesting that the downregulation was due to mechanisms other than methylation of the promoter region. These findings suggest that RUNX1 and CBFB in addition to RUNX3 play some roles in gastric cancers and that roles of RUNX gene family in gastric cancer are more widespread and complex than previously realized.