细胞膜GRP78蛋白通过激活STAT3促进乳腺癌细胞增殖和迁移

发布时间：2018-04-18 12:59

本文选题：sGRP78 + STAT3　；参考：《武汉大学》2015年博士论文

【摘要】：目的：近年来,我国乳腺癌发病率逐年上升,已跃升为女性肿瘤的第一位。耐药乳腺癌复发及转移仍然是无法治愈的疾病,潜伏的肿瘤细胞带来的肿瘤复发和远处转移则是乳腺癌患者死亡的主要原因。已有研究证实,细胞膜高表达GRP78蛋白与癌细胞增殖、迁移、侵袭和治疗耐药有关。然而,其具体的作用机制并不清楚。本文将探讨人乳腺癌组织细胞膜GRP78(surface GRP78,sGRP78)蛋白的表达与疾病进展的关系,并在体外进一步研究sGRP78蛋白促进乳腺癌细胞增殖和迁移的分子机制。方法：利用免疫荧光双标法检测乳腺癌组织和及其癌旁对照组织sGRP78蛋白,Western blot法检测蛋白和蛋白磷酸化的表达,并分析其与临床病理参数的关系。采用携带目的基因腺病毒载体在低表达GRP78蛋白的人乳腺癌细胞MCF-7和MDA-MB-453建立过表达模型；采用靶向STAT3的shRNA慢病毒下调STAT3的表达。分析sGRP78蛋白表达与内质网应激的关系,然后分别应用MTT和TUNEL检测细胞增殖和凋亡,wound healing assay检测细胞迁移,最后探讨sGRP78对JAK2/STAT3信号通路的影响。结果：(1)sGRP78在人乳腺癌组织中有表达,且表达与乳腺癌的病理分期显著相关,分期越高,sGRP78蛋白的表达越强。(2)Ad/GRP78转染细胞可显著提高sGRP78在乳腺癌细胞中的表达。(3)sGRP78蛋白上调不能引起内质网应激；但内质网应激能够提高GRP78蛋白表达并上调GRP78蛋白在细胞膜的分布。(4)sGRP78蛋白过表达促进MCF-7乳腺癌细胞增殖、迁移。(5)sGRP78激活JAK2/STAT3信号通路。(6)STAT3介导sGRP78对乳腺癌细胞的增殖、凋亡、迁徙的影响。结论：我们证实,(1)人乳腺癌组织表达sGRP78蛋白,且与乳腺癌进展相关；(2)sGRP78蛋白通过激活JAK2/STAT3信号通路促进乳腺癌细胞增殖和迁移。
[Abstract]:Objective: in recent years, the incidence of breast cancer in China has increased year by year.Recurrence and metastasis of drug-resistant breast cancer is still an incurable disease. Recurrence and distant metastasis caused by latent tumor cells are the main causes of death in breast cancer patients.It has been proved that the high expression of GRP78 protein on cell membrane is related to the proliferation, migration, invasion and drug resistance of cancer cells.However, its specific mechanism is not clear.In this paper, we will investigate the relationship between the expression of GRP78(surface GRP78) and the progress of the disease in human breast cancer tissue, and further study the molecular mechanism of sGRP78 protein promoting the proliferation and migration of breast cancer cells in vitro.Methods: the expression of protein and protein phosphorylation in breast cancer tissues and its adjacent control tissues were detected by double immunofluorescence blot, and the relationship between protein phosphorylation and clinicopathological parameters was analyzed.The overexpression model was established by using adenovirus vector carrying target gene in MCF-7 and MDA-MB-453 cells with low expression of GRP78 protein, and the expression of STAT3 was down-regulated by shRNA lentivirus targeting STAT3.The relationship between the expression of sGRP78 protein and endoplasmic reticulum stress was analyzed. Then MTT and TUNEL were used to detect cell proliferation and apoptosis healing assay to detect cell migration. Finally, the effect of sGRP78 on JAK2/STAT3 signaling pathway was discussed.Results the expression of SGRP78 in human breast cancer was significantly correlated with the pathological stage of breast cancer.The higher the stage, the stronger the expression of sGRP78 protein. Ad-r-GRP78 transfection cells could significantly increase the expression of sGRP78 in breast cancer cells. The up-regulation of sGRP78 protein could not induce endoplasmic reticulum stress.However, endoplasmic reticulum stress could increase the expression of GRP78 protein and up-regulate the distribution of GRP78 protein on the cell membrane. The overexpression of GRP78 protein promoted the proliferation of MCF-7 breast cancer cells. Migration. 5% sGRP78 activated the JAK2/STAT3 signaling pathway. SGRP78 mediated the proliferation and apoptosis of breast cancer cells.The effects of migration.Conclusion: sGRP78 protein is expressed in human breast cancer tissues, and it is associated with the progression of breast cancer, which promotes the proliferation and migration of breast cancer cells by activating JAK2/STAT3 signaling pathway.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R737.9

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