内质网应激相关蛋白调控自噬在双硫仑诱导胰腺癌细胞凋亡中的作用及机制研究

The efficacy of chemotherapeutic drugs, which is widely used in clinical treatment, is not satisfactory. Our previous work revealed that: the alcohol-abuse drug disulfiram can obviously inhibit the proliferation of pancreatic cancer cells. The autophagy and autophagy-mediated cell death of cells can be observed in DSF treated pancreatic cancer cells. In the next study, we found that the autophagy initiated via endoplasmic reticulum stress induced by DSF, the autophagy decreased in endoplasmic reticulum stress protein inhibit pretreated cells. Based upon these founding, we will investigate the mechanism of the endoplasmic reticulum stress related proteins, including p8﹑sXBP1﹑IREα﹑ATF4﹑CHOP﹑p97﹑p47 involved in ER autophagy induced by DSF. Our study including two parts: First, the functional characterization of the p8-ATF4-CHOP pathway involved in the autophagosome formation. And the role of the p97-p47-VCIP135 complex involved in the autolysome maturation. SiRNA interference, eukaryotic gene overexpression, retrovirus packaging and infection,FACS, cell immunofluorescence and Co-immunoprecipitation techniques will be used in our project, including vitro and vivo treatment. The implementation of this project may provide new evidence in pancreatic cancer treatment with DSF, a highly efficient and inexpensive chemotherapy drug.

目前广泛应用于胰腺癌临床治疗的化疗药物疗效不佳。申请者前期研究发现，临床上常用的戒酒药双硫仑可以明显抑制胰腺癌细胞的增殖，DSF可通过上调胰腺癌细胞的自噬，引起细胞的凋亡，且有证据提示DSF诱导胰腺癌细胞的内质网应激反应与细胞的自噬发生相关，抑制内质网应激则抑制了细胞自噬。基于此，项目申请者拟通SiRNA干扰、真核基因过表达、逆转录病毒包装转染、流式细胞术、细胞免疫荧光及免疫共沉淀等技术，结合体外和体内实验，研究p8-ATF4-CHOP通路在自噬体形成及p97-p47-VCIP135复合物在自噬溶酶体形成过程中的作用，探讨内质网应激相关蛋白p8﹑sXBP1﹑IREα﹑ATF4﹑CHOP﹑p97﹑p47等在DSF诱导的胰腺癌自噬中的调控机制。本项目的实施，可为胰腺癌的临床治疗提供一种高效廉价的化疗药物，并为其提供新的理论基础和实验依据。

目前广泛应用于胰腺癌临床治疗的化疗药物疗效不佳。申请者前期研究发现，临床上常用的戒酒药双硫仑可以明显抑制胰腺癌细胞的增殖，DSF可通过上调胰腺癌细胞的自噬，引起细胞的凋亡，且有证据提示DSF诱导胰腺癌细胞的内质网应激反应与细胞的自噬发生相关，抑制内质网应激则抑制了细胞自噬。基于此，项目申请者拟通SiRNA干扰、真核基因过表达、逆转录病毒包装转染、流式细胞术、细胞免疫荧光及免疫共沉淀等技术，结合体外和体内实验，研究p8-ATF4-CHOP通路在自噬体形成及p97-p47-VCIP135复合物在自噬溶酶体形成过程中的作用，探讨内质网应激相关蛋白p8﹑sXBP1﹑IREα﹑ATF4﹑CHOP﹑p97﹑p47等在DSF诱导的胰腺癌自噬中的调控机制。本项目的实施，可为胰腺癌的临床治疗提供一种高效廉价的化疗药物，并为其提供新的理论基础和实验依据。