胰腺导管腺癌和肿瘤微环境之间相互作用的系统研究：临床病理研究

Microenvironment may play a dual-role in tumor invasion and metastasis: there are both pro- and anti-tumorigenesis factors in the microenvironment of the cancer. Characterization of these factors and understanding their molecular mechanisms are pivotal to cancer diagnosis, prognosis and therapy. Pancreatic ductal adenocarcinoma (PDAC) is a cancer with the worst prognosis. Importantly, PDAC is characterized by having a rich stroma, suggesting intimate interactions between individual cancer cells with its microenvironment. While several recent studies have established the importance of microenvironment for other adenocarcinoma, few studies have focused on the tumor-stroma interaction in pancreatic cancer. We hypothesize that microenvironmental elements, in particular inflammatory cells such as mast cells, may establish a pro-growth environment for cancer cells by promoting angiogenesis. The balance between this impact and immune cells' general anti-cancer activity determines the aggressiveness of the cancer. In this study, we propose to comprehensively characterize the distribution and subtypes of mast cells and blood vessels in four distinct areas of PDAC: the non-invasive areas including peritumoral border and central zones; the invasive front, or the intratumor border zone; and the complete tumor zone, or the intratumor central zone. We will use HE staining, masson staining, and immunohistochemistry analysis to evaluate infiltration of inflammatory cells, fibrosis formation, angiogenesis and lymphoangiogenesis. Comparisons of the localization and zone-specific distribution of mast cells using cox-hazard ratio model will provide novel prognostic. We also propose to dissect the molecular mechanism and signaling pathways that mediate mast cells' capacity to promote tumorigenesis by a combined molecular biology and cell biology strategy. Ultimately, our proposed study will provide significant and novel insight into the mechanism and therapeutic of PDAC.

肿瘤微环境具有促肿瘤和抑肿瘤双重作用。胰腺癌间质含量丰富，肿瘤细胞和细胞间质的相互作用对于肿瘤侵袭转移复发可能具有重要影响。目前对胰腺癌微环境研究较少，缺乏按照空间定位系统评价胰腺癌微环境的临床研究。本研究设想肿瘤微环境如肥大细胞等炎性细胞可能对于肿瘤细胞的血管生成具有促进作用，与免疫细胞抗癌作用的平衡决定癌症的进展：将胰腺癌标本分为肿瘤中心区、肿瘤边缘区、癌旁边缘区和癌旁中心区四个部分，通过HE染色、Masson染色、免疫组化染色评价其炎细胞浸润数量、纤维化程度、微血管和微淋巴管数量等炎症反应情况，结合病理信息进行统计分析，评价胰腺癌微环境中炎症反应的临床意义；探讨肥大细胞的区域分布特点，结合术后随访进行多因素生存分析，评价肥大细胞浸润对胰腺癌病人预后的影响；采用分子生物学和细胞生物学研究肥大细胞促进肿瘤生成的分子机制和信号转导途径。期望对胰腺癌机制及其临床治疗的研究具有新的启示。

微环境在肿瘤发生发展中的作用已受到广泛关注。胰腺癌导管腺癌（PDAC)预后差，其组织学特点是间质含量丰富，肿瘤细胞和细胞间质的相互作用对于肿瘤侵袭转移复发可能具有重要影响。目前对胰腺癌微环境研究较少，缺乏按照空间定位系统评价胰腺癌微环境的临床研究。本研究设想肿瘤微环境如肥大细胞等炎性细胞可能对于肿瘤细胞的血管生成具有促进作用，与免疫细胞抗癌作用的平衡决定癌症的进展：将胰腺癌标本分为肿瘤中心区、肿瘤边缘区、癌旁边缘区和癌旁中心区四个部分，通过HE 染色、Masson 染色、免疫组化染色评价其炎细胞浸润数量、纤维化程度、微血管和微淋巴管数量等炎症反应情况，结合病理信息进行统计分析，评价胰腺癌微环境中炎症反应的临床意义；探讨肥大细胞的区域分布特点，结合术后随访进行多因素生存分析，评价肥大细胞浸润对胰腺癌病人预后的影响, 为临床提供新的预后指标。本课题执行期内，收集了经病理确诊的PDAC病例30例和正常人5例，取胰腺癌和正常胰腺组织及其血清，按课题计划对PDAC肿瘤的4个部位进行组织学和免疫组化染色，其中用类胰蛋白酶/糜蛋白酶显示肿瘤间质中的肥大细胞，血管内皮细胞标志CD34显示毛细血管网分布，同时用Masson三色染色显示细胞外基质中的纤维成分。根据时间节点的病例随访资料尚待收集。初步结果显示，肿瘤边缘区肥大细胞数量最多，主要以T型（表达胰蛋白酶）为主，同时此部位也是CD34阳性内皮细胞显示血管分布最丰富的区域。对组织标本进行胰蛋白酶元的Western blot检测也得出类似结果。肿瘤边缘区及侵袭前沿分布较多的肥大细胞并具有较多的血管网提示肥大细胞在PDAC侵袭转移中有重要作用，为进一步揭示肥大细胞促进胰腺肿瘤浸润转移的分子机制和信号转导途径并建立新的胰腺癌预后指标奠定了基础。