Novel differentiation induction therapy for colorectal cancer by targeting to CEACAM1

已知癌胚抗原相关细胞粘附分子1（CEACAM1）在腺瘤和癌中在转录水平下调。最近的研究表明CEACAM1在大肠癌中蛋白水平过表达，并与临床分期相关。结肠直肠癌细胞重新表达CEACAM1的原因尚不清楚。本研究的目的是阐明CEACAM1在结直肠癌中重新表达的意义。用CEACAM1长（CEACAM1-L）或短（CEACAM1-S）胞质结构域特异性抗体对164例结直肠癌患者的临床样本进行免疫组织化学分析。根据CEACAM1再表达的临床意义计算转移和生存的危险因素。采用侵袭小室和伤口愈合试验检测CEACAM1表达对大肠癌细胞侵袭和迁移的影响。CEACAM1-L和CEACAM1-S在浸润前沿染色强度更高， 关于我们 Han位于肿瘤的腔表面。在侵袭前沿观察到长和短胞质亚型表达水平之间的差异。多因素分析显示CEACAM1-L优势是淋巴结转移、血行转移和短生存期的独立危险因素。Kaplan-Meier评估表明CEACAM1-L优势与较短的生存时间相关（p <0.0001）。在侵袭室和伤口愈合测定中，CEACAM1-L促进侵袭和迁移。在结直肠癌的浸润前沿观察到CEACAM1的再表达。CEACAM1-L优势与结直肠癌患者的转移和较短的生存期相关。CEACAM1-L优势对结直肠癌细胞的侵袭和迁移至关重要。组织学未分化癌细胞在肿瘤边缘以外形成的肿瘤出芽与淋巴结转移有关。然而，具有中空的、可想象的肿瘤出芽的组织学晚期表型的癌巢尚未讨论。其次，我们研究了在浸润边缘外存在的“中空球体”的外观，并与转移和预后有关。此外，我们提出CEACAM1亚型平衡与SWH形成相关。用CEACAM1和M30作为凋亡标志物进行免疫组织化学分析，以解决SWH的CEACAM1表达和SWH形成的中央细胞凋亡。对314例结直肠癌患者的浸润边缘外SWH与临床病理特征的关系进行了分析。用CEACAM1 cDNA或shRNA转染的结直肠癌细胞进行三维（3D）培养，以评估CEACAM1同种型平衡控制结直肠SWH形成。M30染色和连续切片CEACAM1染色证实SWH形成伴随中央细胞凋亡。在314例患者中，96例（30.4%）被归类为SWH。SWH是转移和生存期缩短的独立危险因素。在三维培养中，CEACAM1亚型平衡调节结直肠癌细胞SWH的形成，肿瘤边缘以外的SWH对预测结直肠癌的恶性潜能比肿瘤出芽更重要。少

Carcinoembryonic antigen-related cell adhesion molecule 1(CEACAM1) is known to be downregulated at the transcriptional level in adenoma and carcinoma. Recent reports have shown that CEACAM1 is overexpressed at protein level in colorectal cancer, and correlated with clinical stage. The reason why colorectal cancer cells re-expressed CEACAM1 remains unclear. The aim of the present study was to clarify the implication of CEACAM1 re-expression in colorectal cancer. Immunohistochemical analyses were conducted with CEACAM1 long (CEACAM1-L) or short (CEACAM1-S) cytoplasmic domain-specific antibodies on clinical samples from 164 patients with colorectal cancer. The risk factors for metastasis and survival were calculated for clinical implication of CEACAM1 re-expression. Invasion chamber and wound healing assays were performed for the effect of CEACAM1 expression on invasion and migration of colorectal cancer cells. CEACAM1-L and CEACAM1-S stained with greater intensity at the invasion front t … More han at the luminal surface of tumors. Differences between the long and short cytoplasmic isoform expression levels were observed at the invasion front. Multivariate analysis showed that CEACAM1-L dominance was an independent risk factor for lymph node metastasis, hematogenous metastasis, and short survival. The Kaplan-Meier evaluation demonstrated that CEACAM1-L dominance was associated with shorter survival time (p<0.0001). In the invasion chamber and wound healing assays, CEACAM1-L promoted invasion and migration. Re-expression of CEACAM1 is observed at the invasion front of colorectal cancer. CEACAM1-L dominance is associated with metastasis and shorter survival of the patients with colorectal cancer. CEACAM1-L dominance is important for colorectal cancer cells invasion and migration. Tumor budding formed by histological undifferentiated cancer cells beyond the border of tumor margin is associated with lymph node metastasis. However, cancer nests with a hollow, conceivable histological advanced phenotype of tumor budding, has not been discussed. Next we investigate that the appearance "spheroid with a hollow (SWH)" exists beyond the border of the invasive margin, and associates with metastases and prognosis. Moreover, we present that CEACAM1 isoform balance is associated with SWH formation. Immunohistochemical analyses with CEACAM1 and M30 as an apoptosis marker were performed, to address CEACAM1 expression of SWH and central cells apoptosis for SWH formation. The correlations between the SWH beyond the border of the invasive margin and clinicopathological characteristics of 314 patients with colorectal cancer were evaluated. Three dimensional (3D) culture was conducted with colorectal cancer cells transfected with CEACAM1 cDNA or shRNA, to evaluate that CEACAM1 isoform balance controls colorectal SWH formation. The existence of SWH formation accompanying the central cells apoptosis was confirmed by M30 staining and serial section with CEACAM1 staining. Of the 314 patients, 96 (30.4%) were classified as having SWH. SWH is an independent risk factor for metastases and shorter survival. In 3D culture, CEACAM1 isoform balance modulated SWH formation of colorectal cancer cells.The colorectal cancer SWH beyond the border of tumor margin is more important for prediction of malignant potential than tumor budding. Less