Overexpression of claudin-3 and claudin-4 receptors in uterine serous papillary carcinoma - Novel targets for a type-specific therapy using Clostridium perfringens enterotoxin (CPE)
Overexpression of claudin-3 and claudin-4 receptors in uterine serous papillary carcinoma - Novel targets for a type-specific therapy using Clostridium perfringens enterotoxin (CPE)
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DOI：
10.1002/cncr.22536
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发表时间：
2007-04-01
期刊：
影响因子：
6.2
通讯作者：
Pecorelli, Sergio
中科院分区：
文献类型：
--
作者：
Santin, Alessandro D.;Bellone, Stefania;Pecorelli, Sergio
背景子宫浆液性乳头状癌（USPC）是一种高度侵袭性的子宫内膜癌。使用基因表达谱，我们最近确定了紧密连接蛋白-3和紧密连接蛋白-4受体在有限的USPC中的高表达。这些紧密连接蛋白分别代表细胞毒性产气荚膜梭菌肠毒素（CPE）的低亲和力和高亲和力受体，足以介导CPE结合并引发随后的毒素介导的细胞溶解。探讨了靶向该途径治疗USPC的可能性。在来自20个连续USPC患者的快速冷冻和福尔马林固定的石蜡包埋组织中，在mRNA和蛋白质水平上分析Claudin-3和Claudin-4受体表达。还在体外研究了重组CPE作为针对原发性、转移性和化疗耐药USPC细胞系的新型疗法的潜力。最后，在皮下和腹腔内移植表达claudin-3和claudin-4的USPC的SCID小鼠中，研究亚致死剂量CPE的体内治疗作用。总之，通过定量逆转录酶聚合酶链反应（RT-PCR），检测的100%（20/20）初级快速冷冻USPC过表达1种或2种CPE受体。在大多数通过免疫组织化学检测的USPC标本中记录了膜免疫反应性的claudin-4蛋白表达，而在正常子宫内膜对照组织样品中仅发现低水平的膜染色。当在体外用不同浓度的CPE孵育原发性和转移性短期USPC细胞系时，证实了剂量依赖性细胞毒性效应。在体内，肿瘤内注射耐受性良好剂量的CPE在大型皮下USPC异种移植物中导致所有治疗动物的大面积肿瘤细胞坏死和肿瘤消失，而亚致死腹腔注射CPE对肿瘤进展具有显著的抑制作用，延长了具有化疗耐药的腹腔内USPC癌病的动物的生存期。Claudin-3和claudin-4受体可能为CPE作为一种新型的针对这种高度侵袭性和化疗耐药的子宫内膜癌变体的特异性治疗提供有希望的靶点。
BACKGROUND. Uterine serous papillary carcinoma (USPC) represents a highly aggressive variant of endometrial cancer. Using gene expression profiling, we recently identified high expression of the claudin-3 and claudin-4 receptors in a limited set of USPC. These tight junction proteins represent the low- and high-affinity receptors, respectively, for the cytotoxic Clostridium perfringens enterotoxin (CPE) and are sufficient to mediate CPE binding and trigger subsequent toxin-mediated cytolysis. The potential for targeting this pathway in the treatment of USPC was explored.METHODS. Claudin-3 and claudin-4 receptor expression was analyzed at the mRNA and protein levels in flash-frozen and formalin-fixed, paraffin-embedded tissue from 20 consecutive USPC patients. The potential of recombinant CPE as a novel therapy against primary, metastatic, and chemotherapy-resistant USPC cell lines was also investigated in vitro. Finally, the in vivo therapeutic effect of sublethal doses of CPE was studied in SCID mouse xenografts harboring subcutaneous and intraperitoneal USPC that expressed claudin-3 and claudin-4.RESULTS. In all, 100% (20 out of 20) of the primary flash-frozen USPC tested overexpressed 1 or both CPE receptors by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR). Membranous immunoreactivity for claudin-4 protein expression was documented in the majority of USPC specimens tested by immunohistochemistry, whereas only a low level of membranous staining was found in normal endometrial control tissue samples. When primary and metastatic short-term USPC cell lines were incubated with different concentrations of CPE in vitro, a dose-dependent cytotoxic effect was demonstrated. In vivo, intratumoral injections of well-tolerated doses of CPE in large subcutaneous USPC xenografts led to large areas of tumor cell necrosis and tumor disappearance in all the treated animals, whereas sublethal intraperitoneal injections of CPE had a significant inhibitory effect on tumor progression, with extended survival of animals harboring chemotherapy-resistant intra-abdominal USPC carcinomatosis.CONCLUSIONS. Claudin-3 and claudin-4 receptors may offer promising targets for the use of CPE as a novel type-specific therapy against this highly aggressive and chemotherapy-resistant variant of endometrial cancer.