组胺H4受体调控食管癌细胞系Eca-109的作用及其信号传导机制的研究

Esophageal cancer is a common digestive tract tumor. Around the world there are about 500,000 people died from esophageal cancer every year with our country as the hard-hit area. Looking for a new and effective molecular anticancer target is vital for the treatment and prognosis of esophageal cancer. An accumulating evidence has documented that histamine receptor 4 is widely involved in the the biological behavior of gastrointestinal tumor, including gastric carcinorma, colorectal carcinorma and so on. However, whether histamine receptor 4 can regulate the proliferation, apoptosis, metastasis and drug resistance of esophageal carcinorma remain unknown. Our preliminary experiments confirmed that: the expression of histamine receptor 4 significantly decreased in several esophageal carcinorma cell lines compared with normal esophageal epithelial cell line SHEE. And the agonist of histamine receptor 4 (4-methylhistamine) significantly inhibited proliferation and epithelial mesenchymal transformation of esophageal carcinorma cell line Eca-109, as well as the transplanted tumor growth. Based on these findings, this research is proposed to study the expression of histamine receptor 4 in the esophageal carcinorma tissue and cell lines, to verify the function of histamine receptor 4 in the proliferation, apoptosis, metastasis and drug resistance of the esophageal cancer cell line Eca-109, and to explore the potiential signaling pathways underlying the histamine receptor 4. The results is expected to provide new academic information to recognize the anti-tumor effect of histamine receptor 4 and also can provide new medical target for clinical treatment of esophageal cancer.

食管癌是一种常见的消化道肿瘤，全世界每年约有50万人死于食管癌，我国是世界上食管癌高发地区之一，寻找一种新的有效的分子调控靶点对于食管癌的治疗和预后至关重要。大量研究证实组胺H4受体参与多种消化系肿瘤的行为学的调控，但是H4受体在食管癌中的作用目前为止尚无文献报道。我们的预实验结果提示：多种食管癌细胞系中H4受体表达紊乱，并且激活H4受体可抑制食管癌细胞系Eca-109的增殖、上皮间质转化过程以及多条与食管癌生物学行为密切相关信号通路的活化。在此基础上，本项目拟研究H4受体在食管癌组织和食管癌细胞系中的表达；并探讨H4受体对于食管癌细胞系Eca-109的增殖、凋亡、转移和耐药的调控作用；并验证其中可能的信号通路。研究结果可望为认识H4受体的抗肿瘤作用提供新的学术资料，也可为临床治疗食管癌提供新的药物靶点。

食管癌是一种常见的消化道肿瘤，全世界每年约有50万人死于食管癌，我国是世界上食管癌高发地区之一，寻找一种新的有效的分子调控靶点对于食管癌的治疗和预后至关重要。大量研究证实组胺H4受体参与多种消化系肿瘤的行为学的调控，但是H4受体在食管癌中的作用目前为止尚无文献报道。.我们通过构建临床食管癌组织标本样品库，首次证实组胺受体4（H4R）在食管正常组织和食管鳞癌中的表达，分子生物学结果提示H4R部分食管鳞癌组织中的表达出现缺失。我们构建了TE-1细胞系的裸鼠移植瘤模型，分组每日腹腔注射给予生理盐水、H4R的激动剂4MH、阻断剂JNJ和4MH+JNJ，观察移植瘤的在体生长情况并记录裸鼠的存活时间，结果证实：激动剂4MH激动H4R后，裸鼠的成瘤大小得到显著的抑制，同时裸鼠成瘤后裸鼠的生存期时间明显延长；而阻断剂JNJ处理后则显著提高裸鼠的成瘤大小。在测序实验中发现H4R可以调控MAPK条目和PATHWAY中的MAPK抑制条目中的差异基因做GO条目注释，结果提示H4R的激动能够显著抑制多条和细胞增殖相关的条目。将各个条目中的差异基因进行PCR验证，结果证实：激动H4R可以显著抑制 JUN、TGF-β2、TGF-β1、RASGRF1 、EGF、HSPA1B、HSPA1A 的表达，阻断H4R则可以显著促进这些基因的表达。进一步加使用ERK阻断剂则仅仅抑制了由于JNJ阻断H4R所提高的HSP70和TGFb1、2，说明HSP70和TGFb1、2是H4R-ERK信号依赖性的下游分子。初步使用HSP70抑制剂和TGFb的siRNA处理后，可以抑制H4R分子阻断剂对于食管鳞癌的促增殖作用，说明HSP70和TGFb1、2是H4R重要的调控食管鳞癌肿瘤增殖的相关分子。研究结果为认识H4受体的抗肿瘤作用提供新的学术资料，也可为临床治疗食管癌提供新的药物靶点。