重组新城疫病毒rL-RVG通过乙酰胆碱受体诱导胃癌细胞内质网应激-自噬-凋亡的机制研究

Newcastle disease virus named as NDV has wide application prospect in cancer therapy, but the study of recombinant Newcastle disease virus used to treat gastric cancer is very poor. The recombinant Newcastle disease virus (rL-RVG) expressing stably the rabies virus glycoprotein, which has the safety, high immune activity, constructed by the Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences was used. Previous studies showed that rL-RVG could induce apoptosis in gastric cancer cells, significantly improved the level of expression of autophagy and endoplasmic reticulum stress related proteins compared with the wild type virus, and we found that the expression of rL-RVG-induced apoptosis and autophagy related proteins decreased in gastric cancer cells after pretreatment with the autophagy pathway inhibitor 3-MA. On the other hand ,we demonstrated that the alpha 7-nAChR expressed in gastric cancer cell lines ,which prompted that rL-RVG maybe enhance the apoptosis of tumor cells through blocking nAChR channel. In this study, we will infect human gastric adenocarcinoma cells with rL-RVG in vitro, and pretreat with the inhibitors of nAChR pathway and endoplasmic reticulum stress pathways,then detect nAChR pathway, endoplasmic reticulum stress, autophagy and apoptosis related proteins. Further, we will give the tumor bearing rat intervention with the recombinant Newcastle disease virus (rL-RVG) in vivo after the gastric cancer orthotopic transplantation models have been constructed. At last, the relationship between nAChR expression levels and the clinical features in patients with gastric cancer will be analyzed.All the above researches aim at elucidating the mechnism of the recombinant Newcastle disease virus rL-RVG inducing endoplasmic reticulum stress-autophagy-apoptosis-oncolytic through blocking the nAChR pathway in stomach adenocarcinoma, it will provide molecular bases for developing rL-RVG into a drug candidate for the treatment of gastric carcinoma.

新城疫病毒NDV在肿瘤治疗中具有广泛的应用前景，目前重组NDV用于胃癌治疗的研究甚少。我们采用中国农业科学院哈尔滨兽医研究所构建的安全、高免疫活性、稳定表达狂犬病毒糖蛋白的新城疫病毒疫苗rL-RVG，前期研究发现rL-RVG较野生型病毒明显促进胃癌细胞凋亡，且自噬及内质网应激蛋白水平显著提高，并发现抑制自噬后能抑制凋亡；α7-nAChR在胃癌细胞系较多表达，rL-RVG可能通过阻断nAChR途径促进肿瘤细胞凋亡。本研究采用rL-RVG体外感染人胃腺癌细胞，并予以nAChR途径及内质网应激通路干预，检测nAChR途径蛋白及内质网应激、自噬、凋亡水平，同时进行体内荷瘤鼠重组病毒感染干预研究，并分析nAChR表达与胃癌临床特征的关系，旨在阐明重组新城疫病毒rL-RVG通过拮抗nAChR途径诱导内质网应激-自噬-凋亡-溶瘤的机制，为重组新城疫病毒rL-RVG用于临床治疗胃癌提供理论依据。

新城疫病毒NDV在肿瘤治疗中具有广泛的应用前景，目前重组NDV用于胃癌治疗的研究甚少。我们采用中国农业科学院哈尔滨兽医研究所构建的安全、高免疫活性、稳定表达狂犬病毒糖蛋白的新城疫病毒疫苗rL-RVG，前期研究发现rL-RVG较野生型病毒明显促进胃癌细胞凋亡且自噬及内质网应激蛋白水平显著提高，并发现抑制自噬后能抑制凋亡；推测rL-RVG可能通过阻断nAChR途径促进肿瘤细胞凋亡。本研究采用rL-RVG体外感染人胃腺癌细胞(SCG、HGC、BGC)，并予以nAChR途径及内质网应激通路抑制剂进行干预，检测nAChR途径蛋白及内质网应激、自噬、凋亡水平，且进行干预后胃癌细胞的迁移及相关通路蛋白的影响，同时进行重组病毒感染后体内成瘤研究，并分析nAChR表达与胃癌临床特征的关系，结果发现：rL-RVG体内外抑制人胃腺癌细胞的生长，体外研究发现能促进内质网应激、自噬及其凋亡，并抑制其迁移，临床研究发现α7-nAChR、miR-125b-5p在胃癌组织较多表达，并与临床特征密切相关，故本实验进一步阐明重组新城疫病毒rL-RVG通过拮抗nAChR途径诱导内质网应激-自噬-凋亡的机制，为重组新城疫病毒rL-RVG用于临床治疗胃癌提供理论依据。