基于微流控芯片的CAFs影响胃癌细胞耐药及GRP-78表达相关性

The Relevance of CAFs Influencing the Drug Resistant of Gastric Cancer Cells and the GRP-78 Expression Based on Microfluidic Chip

目的:利用微流控芯片平台探索肿瘤相关成纤维细胞(CAFs)影响人胃癌SGC7901细胞对化疗药物的耐药及其与GRP78表达的相关性,从而为临床工作提供以CAFs和GRP78为靶点来改善肿瘤耐药的新思路。方法:本实验自主设计了一个可用于药敏检测及蛋白半定量的三维联合共培养微流控芯片体系,通过灌注细胞悬液-凝胶混合物于三维培养池内来实现胃癌细胞系的三维培养。应用凋亡染色方法检测SGC7901细胞单独培养组(对照组)、SGC7901细胞+NFs共培养组、SGC7901细胞+CAFs共培养组中SGC7901细胞对不同浓度的化疗药物顺铂的药物敏感情况。应用免疫荧光法检测对照组和实验组SGC7901细胞GRP78表达。结果:SGC7901细胞+CAFs共培养组与单培养组相比,SGC7901细胞对顺铂的敏感性较SGC7901细胞单独培养时低(P<0.05),SGC7901细胞+CAFs共培养组SGC7901细胞中GRP78蛋白表达较对照组升高(P<0.05)。结论:CAFs可以诱导人胃癌SGC7901细胞对顺铂耐药;CAFs可能通过上调人胃癌SGC7901细胞中GRP78的表达而导致耐药。

Objective：The platform of microfluidic chip is used to explore the relevance of cancerassociated fibroblasts（CAFs） influencing the resisitence of human gastric cancer SGC7901 cells to chemotherapy drugs and the expression of GRP-78,to provide the new idea that CAFs and GRP78 as targets improve tumor drug resistance for clinical work.Method：A microfluidic chip system of three-dimensional co-culture was designed independently which could be used for drug sensitive detection and protein semi-quantitative in this experiment.Three-dimensional culture of gastric cancer cell line was achieved by perfusing cell suspension-gel mixture.Apoptosis staining was applied to detect the sensitivity of SGC7901 cells to chemotherapeutic drugs cisplatin with different concentrations in separate culture group of SGC7901 cells（the control group）,co-culture group of SGC7901 cells and NFs,co-culture group of SGC7901 cells and CAFs.The expressions of GRP78 in the control group and experimental group were detected by immunofluorescent assay.Result：Compared with the control group,the sensitivity of human gastric cancer SGC7901 cells to cisplatin in the co-culture group of SGC7901 cells and CAFs was obviously lower than that in the control group（P〈0.05）.Compared with the control group,the GRP78 expressions of SGC7901 cells in the co-culture group of SGC7901 cells and CAFs was distinctly higher（P〈0.05）.Conclusion：CAFs can induce the drug resistance of SGC7901 cells to cisplatin; CAFs may lead to drug resistance by up-regulating the GRP78 expression in human gastric cancer SGC7901 cells.