Fibronectin 1促进胃癌细胞耐药及其作用机制研究  

Fibronectin 1 promotes drug resistance of gastric cancer cells and its mechanism

作　　者：岳芙蓉 彭科瑜 张力 张军 YUE Furong;PENG Keyu;ZHANG Li;ZHANG Jun(Department of Oncology,Chongqing University Three Gorges Hospital,Chongqing,404100,China)

机构地区：[1]重庆大学附属三峡医院肿瘤科,重庆404100

出　　处：《肿瘤药学》2023年第1期43-49,共7页Anti-Tumor Pharmacy

基　　金：重庆市自然科学基金面上项目(cstc2017jcyj-mxmX3527)。

摘　　要：目的探讨纤连蛋白1(FN1)促进胃癌细胞耐药及其作用机制。方法选择胃癌细胞系MKN45、MKN45/DDP、MKN45/VCR进行pLV-FN1、pLV-NC、FN1 siRNA慢病毒和脂质体3000瞬时转染,将转染pLVFN1的设为FN1过表达组(FN1),转染pLV-NC的为阴性对照组(Vector),转染FN1 siRNA慢病毒的为FN1敲低组(si-FN1),转染脂质体3000的为空白对照组(si-control)。分别采用qRT-PCR、MTT、克隆形成实验、Western blotting检测FN1 mRNA和蛋白的表达,MKN45/DDP细胞增殖、凋亡、集落形成情况及耐药相关蛋白的表达。结果FN1 mRNA和蛋白在MKN45和MKN45/VCR细胞中的表达水平均显著低于MKN45/DDP细胞(P<0.05);FN1敲低组细胞增殖能力显著低于空白对照组(P<0.001),FN1过表达组细胞增殖能力明显高于阴性对照组(P<0.001);FN1敲低组细胞凋亡率明显高于空白对照组(P<0.001),FN1过表达组细胞凋亡率显著低于阴性对照组(P<0.05);FN1敲低组细胞集落形成能力明显低于空白对照组(P<0.001),FN1过表达组细胞集落形成能力显著高于阴性对照组(P<0.001);FN1敲低组MRP-1、p-STAT3、STAT3、p-AKT蛋白表达水平均明显低于空白对照组(P<0.001)。结论FN1的高表达可能参与胃癌化疗耐药的相关过程,下调FN1表达有助于恢复耐药胃癌细胞对化疗的敏感性,有望为改善胃癌化疗疗效提供新的思路和方法。Objective To investigate the mechanism of fibronectin 1(FN1)in promoting drug resistance of gastric cancer cells.Methods MKN45 gastric cancer cell line,MKN45/DDP and MKN45/VCR cell lines were respectively transfected with pLV-FN1,pLV-NC,FN1 siRNA,and lipofectin 3000,and were taken as the FN1 overexpression group(FN1),negative control group(Vector),FN1 knockdown group(si-FN1),and the blank control group(si-control),respectively.The expression of FN1 mRNA and protein,the proliferation,apoptosis,colony formation of cells,and expression of drug resistance-related proteins in MKN45/DDP cells were respectively detected by qRT-PCR,MTT assay,colony formation test and Western blotting.Results The expression of FN1 mRNA and protein in MKN45 and MKN45/VCR cells was significantly lower than in MKN45/DDP cells(P<0.001).The cell proliferation ability in the FN1 knockdown group was significantly lower than in the blank control group(P<0.05),but it was significantly higher in the FN1 overexpression group than in the negative control group(P<0.001).The cell apoptosis rate in the FN1 knockdown group was significantly higher than in the blank control group(P<0.001),but it was significantly lower in the FN1 overexpression group than in the negative control group(P<0.001).The colony formation ability of the FN1 knockdown group was significantly lower than that of blank control group(P<0.05),but that of the FN1 overexpression group was significantly higher than that of negative control group(P<0.001).The expression levels of the proteins MRP-1,p-STAT3,STAT3 and p-Akt in FN1 knockdown group were significantly lower than those in blank control group(P<0.001).Conclusion The high expression of FN1 may be involved in the related process of chemotherapy resistance in gastric cancer.The down-regulation of FN1 expression can reduce the resistance of gastric cancer cells to chemotherapy.This may provide new ideas and methods for further improvement of chemotherapy in gastric cancer.

关 键 词：FN1 胃癌 耐药 增殖 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]