Divergent effects of muscarinic receptor subtype gene ablation on murine colon tumorigenesis reveals association of M3R and zinc finger protein 277 expression in colon neoplasia.
Divergent effects of muscarinic receptor subtype gene ablation on murine colon tumorigenesis reveals association of M3R and zinc finger protein 277 expression in colon neoplasia.
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DOI：
10.1186/1476-4598-13-77
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发表时间：
2014-04-03
期刊：
影响因子：
37.3
通讯作者：
Raufman JP
中科院分区：
文献类型：
--
作者：
Cheng K;Xie G;Khurana S;Heath J;Drachenberg CB;Timmons J;Shah N;Raufman JP
M3 和 M1 亚型毒蕈碱受体在正常和肿瘤性肠上皮细胞中共表达。在小鼠中，消除 Chrm3（编码 M3R 的基因）可显着减弱肠道肿瘤的形成。在这里，我们研究了 Chrm1 基因消融单独以及与 Chrm3 消融结合的效果。我们使用野生型 Chrm1-/-、Chrm3-/- 和联合 Chrm1-/-/Chrm3-/- 敲除（双敲除）小鼠。用氧化偶氮甲烷（一种肠道选择性致癌物）治疗动物。 20 周后，对结肠肿瘤进行计数并通过组织学和免疫组织化学染色进行分析。使用微阵列分析肿瘤基因表达，并通过 RT-PCR 验证结果。通过分析人类结肠癌和邻近正常结肠组织中的基因和蛋白质表达，扩展了主要发现。与野生型小鼠相比，氧化偶氮甲烷处理的 Chrm3-/- 小鼠的结肠肿瘤更少且更小。在 Chrm1-/- 或双敲除小鼠中未观察到结肠肿瘤数量和大小的减少。为了获得对这些不同表型的遗传洞察，我们使用无偏差的微阵列方法来比较 Chrm3-/- 肿瘤中的基因表达与野生型小鼠中的基因表达。我们检测到 430 个基因的表达发生改变，并通过定量 RT-PCR 验证了前 14 个上调基因和 14 个下调基因。比较野生型、Chrm3-/-、Chrm1-/- 和双敲除小鼠肿瘤中这 28 个基因子集的表达，发现 M3R 缺陷型和双敲除小鼠组织中编码锌指蛋白 277 的 Zfp277 表达显着降低，Zfp277 蛋白表达也发生平行变化。值得注意的是，与邻近的正常结肠相比，人类结肠癌中 ZNF277（Zfp277 的人类类似物）的 mRNA 和蛋白质增加，同时 M3R 表达也发生平行变化。我们的结果确定了一个新的候选小鼠基因 Zfp277，其表达模式与介导 Chrm3 和 Chrm1 基因消融对小鼠肠道肿瘤的不同作用的作用相一致。通过发现人类结肠癌中 ZNF277 的表达增加以及 M3R 表达的平行增加，这一观察结果的生物学重要性得到了加强。锌指蛋白277在结肠癌中的作用及其与M3R表达和激活的关系值得进一步研究。
M3 and M1 subtype muscarinic receptors are co-expressed in normal and neoplastic intestinal epithelial cells. In mice, ablating Chrm3, the gene encoding M3R, robustly attenuates intestinal tumor formation. Here we investigated the effects of Chrm1 gene ablation, alone and in combination with Chrm3 ablation. We used wild-type, Chrm1-/-, Chrm3-/- and combined Chrm1-/-/Chrm3-/- knockout (dual knockout) mice. Animals were treated with azoxymethane, an intestine-selective carcinogen. After 20 weeks, colon tumors were counted and analyzed histologically and by immunohistochemical staining. Tumor gene expression was analyzed using microarray and results validated by RT-PCR. Key findings were extended by analyzing gene and protein expression in human colon cancers and adjacent normal colon tissue. Azoxymethane-treated Chrm3-/- mice had fewer and smaller colon tumors than wild-type mice. Reductions in colon tumor number and size were not observed in Chrm1-/- or dual knockout mice. To gain genetic insight into these divergent phenotypes we used an unbiased microarray approach to compare gene expression in tumors from Chrm3-/- to those in wild-type mice. We detected altered expression of 430 genes, validated by quantitative RT-PCR for the top 14 up- and 14 down-regulated genes. Comparing expression of this 28-gene subset in tumors from wild-type, Chrm3-/-, Chrm1-/- and dual knockout mice revealed significantly reduced expression of Zfp277, encoding zinc finger protein 277, in tissue from M3R-deficient and dual knockout mice, and parallel changes in Zfp277 protein expression. Notably, mRNA and protein for ZNF277, the human analogue of Zfp277, were increased in human colon cancer compared to adjacent normal colon, along with parallel changes in expression of M3R. Our results identify a novel candidate mouse gene, Zfp277, whose expression pattern is compatible with a role in mediating divergent effects of Chrm3 and Chrm1 gene ablation on murine intestinal neoplasia. The biological importance of this observation is strengthened by finding increased expression of ZNF277 in human colon cancer with a parallel increase in M3R expression. The role of zinc finger protein 277 in colon cancer and its relationship to M3R expression and activation are worthy of further investigation.
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影响因子：
4.1
作者：
Boss, A;Oppitz, M;Drews, U
通讯作者：
Drews, U
影响因子：
5.6
作者：
Jiménez, E;Montiel, M
通讯作者：
Montiel, M
影响因子：
3.6
作者：
Gautam, D;Heard, TS;Wess, J
通讯作者：
Wess, J
DOI：
10.1016/j.bbrc.2005.07.031
发表时间：
2005-09-09
影响因子：
3.1
作者：
Rimmaudo, LE;de la Torre, E;Sales, ME
通讯作者：
Sales, ME
癌症的分子起源：结直肠癌的分子基础。
DOI：
10.1056/nejmra0804588
发表时间：
2009-12-17
期刊：
The New England journal of medicine
影响因子：
--
作者：
Markowitz SD;Bertagnolli MM
通讯作者：
Bertagnolli MM