细胞间黏附分子-1 介导鼠结肠腺癌的侵袭。
Intercellular adhesion molecule-1 mediates murine colon adenocarcinoma invasion.
机构信息
Department of Surgery, Denver Health Medical Center, University of Colorado, Denver Campus, Denver, Colorado.
Department of Surgery, Denver Health Medical Center, University of Colorado, Denver Campus, Denver, Colorado.
出版信息
J Surg Res. 2014 Mar;187(1):19-23. doi: 10.1016/j.jss.2013.11.001. Epub 2013 Nov 8.
BACKGROUND
Intercellular adhesion molecule-1 (ICAM-1) modulates cell-cell adhesion and is a receptor for cognate ligands on leukocytes. Upregulation of ICAM-1 has been demonstrated in malignant transformation of adenomas and is associated with poor prognosis for many malignancies. ICAM-1 is upregulated on the invasive front of pancreatic metastases and melanomas. These data suggest that the upregulated ICAM-1 expression promotes malignant progression. We hypothesize that the downregulation of ICAM-1 will mitigate tumor progression.
METHODS
Mouse colon adenocarcinoma cells (MC38) were evaluated for the expression of ICAM-1 using Western immunoblot analysis. Short hairpin RNA (shRNA) transduction was used to downregulate ICAM-1. Tumor invasion determined via a modified Boyden chamber was used as a surrogate of tumor progression examining MC38 cells, MC38 ICAM-1 knockdowns, and MC38 transduced with vehicle control. The cells were cultured in full media for 24 h and serum-starved for 24 h. A total of 5 × 10(4) cells were plated and allowed to migrate for 24 h using full media with 10% fetal bovine serum as a chemoattractant. Inserts were fixed and stained with crystal violet. Blinded investigators counted the cells using a stereomicroscope. Statistical analysis was performed by analysis of variance with Fischer protected least significant difference and a P value of <0.05 was considered statistically significant.
RESULTS
ICAM-1 was constitutively expressed on MC38 cells. Transduction with anti-ICAM-1 shRNA vector downregulated ICAM-1 protein expression by 30% according to the Western blot analysis (P < 0.03) and decreased ICAM-1 messenger RNA expression by 70% according to the reverse transcription-polymerase chain reaction. shRNA knockdown cells had a significant reduction in invasion >45% (P < 0.03). There were no significant differences between the invasion rates of MC38 and MC38 vehicle controls.
CONCLUSIONS
Downregulation of ICAM-1 mitigates MC38 invasion. These data suggest that targeted downregulation of tumor ICAM-1 is a potential therapeutic target.
背景
细胞间黏附分子-1（ICAM-1）调节细胞-细胞黏附，是白细胞配体的受体。ICAM-1 的上调已在腺瘤的恶性转化中得到证实，并且与许多恶性肿瘤的预后不良相关。ICAM-1 在胰腺转移瘤和黑色素瘤的侵袭前沿上调。这些数据表明，上调的 ICAM-1 表达促进了恶性进展。我们假设下调 ICAM-1 将减轻肿瘤进展。
方法
使用 Western 免疫印迹分析评估小鼠结肠腺癌细胞（MC38）中 ICAM-1 的表达。短发夹 RNA（shRNA）转导用于下调 ICAM-1。通过改良 Boyden 室测定肿瘤侵袭作为检查 MC38 细胞、MC38 ICAM-1 敲低和转导载体对照的肿瘤进展的替代物。细胞在完全培养基中培养 24 小时并用无血清培养基饥饿 24 小时。将 5×10（4）个细胞接种并在含有 10%胎牛血清的完全培养基中作为趋化剂迁移 24 小时。用结晶紫固定插入物并染色。使用体视显微镜的研究人员进行盲法计数。通过方差分析进行统计分析，并采用 Fischer 保护最小显著差异，P 值<0.05 被认为具有统计学意义。
结果
ICAM-1 在 MC38 细胞中持续表达。根据 Western blot 分析，抗 ICAM-1 shRNA 载体转导下调 ICAM-1 蛋白表达 30%（P<0.03），并根据逆转录-聚合酶链反应降低 ICAM-1 信使 RNA 表达 70%。shRNA 敲低细胞的侵袭率显著降低>45%（P<0.03）。MC38 和 MC38 载体对照的侵袭率之间没有显著差异。
结论
下调 ICAM-1 减轻了 MC38 的侵袭。这些数据表明，靶向下调肿瘤 ICAM-1 是一种潜在的治疗靶点。
Zotero 插件