摘要
目的:探讨细胞外泌体蛋白AGR2表达差异与上皮性卵巢癌获得性耐药的关系。方法:用不同浓度顺铂(6、12、18、24μg/mL)处理细胞,筛选出大鼠卵巢癌耐药细胞株,MTT法检测细胞存活率。用超速差速离心法获得外泌体蛋白,纳米流式检测外泌体蛋白的粒径大小及颗粒浓度。通过qRT-PCR技术比较两组细胞系AGR2相关mRNA的转录情况,并通过Western blot实验对外泌体中AGR2蛋白表达水平进行比较,分析细胞外泌体中蛋白AGR2表达差异与上皮性卵巢癌获得性耐药的关系。结果:NUTU-19肿瘤细胞生存率与化疗药浓度呈负相关;药物浓度的升高与耐药细胞株(NUTU-19R)死亡率相关性不明显。NUTU-19R细胞株AGR2相关mRNA表达水平高于NUTU-19细胞株,差异有统计学意义(P<0.05);NUTU-19R细胞株AGR2蛋白表达高于NUTU-19细胞株,差异有统计学意义(P<0.05)。结论:细胞外泌体蛋白AGR2高表达促进了上皮性卵巢癌获得性耐药的形成。
Objective:To explore the relationship between expression difference of cellular exosome protein AGR2 and acquired drug resistance in epithelial ovarian cancer.Methods:The cells culture in culture medium was conducted with cisplatin(6,12,18,24 μg/mL) to screen out drug-resistant cell lines in rat with ovarian cancer.The survival rate of cells was detected by MTT assay.The exosome protein was obtained by ultrafast differential centrifugation method.The particle size and concentration of exosome protein were detected by nanometer flow cytometry.The transcription of AGR2-related mRNA in the two groups was compared by RT-PCR.The expression of AGR2 protein in exosome was compared by Western Blot to find out the relationship between expression difference of cellular exosome protein AGR2 and acquired drug resistance in epithelial ovarian cancer.Results:MTT assay showed that there was negative correlation between survival rate of tumor cells and the concentration of chemotherapy drugs in NUTU-19 cells.In contrast, the increase of drugs concentration was not proportional to cells death in drug-resistant cell line(NUTU-19R).AGR2-related mRNA in NUTU-19R cell line was higher than that in NUTU-19 cell line(P<0.05),and the expression of AGR2 protein was higher than that in NUTU-19 cell line(P<0.05).Conclusion:The high expression of cellular exosome protein AGR2 promotes the formation of acquired drug resistance in epithelial ovarian cancer.
作者
曹阳
谢鑫
李明珠
CAO Yang;XIE Xin;LI Ming-zhu(Cancer Hospital of China Medical University·Liaoning Cancer Hospital;Liaoning University of Traditional Chinese Medicine,Shenyang 110042,Liaoning,China)
出处
《川北医学院学报》
CAS
2023年第1期15-18,共4页
Journal of North Sichuan Medical College
基金
辽宁省自然科学基金项目(20180550023)。
