解放军医学杂志2025，Vol.50Issue(6)：656-664,9.DOI:10.11855/j.issn.0577-7402.1634.2025.0519
下调Hsa-circ-0101216表达对胰腺癌吉西他滨化疗耐药的影响及其机制
Effect of downregulating Hsa-circ-0101216 expression on gemcitabine chemoresistance in pancreatic cancer and its mechanism
摘要
Abstract
Objective To analyze the effect of Hsa-circ-0101216 on gemcitabine(GEM)chemotherapy resistance in pancreatic cancer and its mechanism.Methods Differentially expressed circRNAs between GEM-resistant pancreatic cancer cells and parent cells were screened using the GEO database.Pancreatic cancer GEM resistant cell lines(BxPC-3-GEM and Capan-1-GEM)were constructed by intermittent concentration gradient method.qRT-PCR was used to detect the expression of Hsa-circ-0101216 in cells.GEM resistant pancreatic cancer cell lines were taken and divided into sh-circ-0101216 group(knockdown of circ-0101216),sh-NC group(transfected with sh-NC),and blank control group(untreated).CCK-8 assay and EdU proliferation assay were used to detect the half inhibitory concentration(IC50)of GEM and proliferation ability of cells in each group.Western blotting was performed to detect the expression of multidrug resistance-related protein 1(MRP1),breast cancer resistance protein(BCRP),and human equilibrative nucleoside transporter-1(hENT-1).A subcutaneous xenograft tumor model of human pancreatic cancer in nude mice was constructed,and sh-NC+GEM group and sh-circ-0101216+GEM group(n=6)were set up.The volume and weight of xenograft tumor in nude mice were compared between the two groups.Western blotting and immunohistochemistry were used to detect the expression of MRP1,BCRP,and hENT-1 proteins in xenograft tumor tissues,and EDU proliferation assay was used to detect the proliferation ability of tumor cells.Results The GEO database screening showed that Hsa-circ-0101216 was up-regulated in GEM-resistant pancreatic cancer cell lines.Pancreatic cancer GEM-resistant cell lines were successfully constructed,and the expression levels of Hsa-circ-0101216 and the IC50 value in GEM-resistant pancreatic cancer cells BxPC-3-GEM and Capan-1-GEM were significantly higher than those in parental cells(P<0.05).In sh-circ-0101216 group,the IC50 values of GEM,cell viability,EdU positivity rate,and the expression levels of MRP1 and BCRP proteins in GEM-resistant pancreatic cancer cells BxPC-3-GEM and Capan-1-GEM were significantly lower than those in blank control group and sh-NC group,while the expression level of hENT-1 protein was significantly higher(P<0.05 or P<0.001).In sh-circ-0101216+GEM group,the weight and volume of subcutaneous xenograft tumors in nude mice,the expression levels and positive expression rates of MRP1 and BCRP proteins in tumor tissues,and the EdU positive rate were significantly lower than those in sh-NC+GEM group,while the expression level and positive expression rate of hENT-1 protein were significantly higher(P<0.05).Conclusions Hsa-circ-0101216 is highly expressed in GEM-resistant pancreatic cancer cell lines.Its knockdown can inhibit the proliferation of pancreatic cancer cells and enhance the chemosensitivity of pancreatic cancer cells to GEM.The mechanism may be related to the regulation of transmembrane transporter protein expression.关键词
Hsa-circ-0101216/胰腺癌/吉西他滨/增殖Key words
Hsa-circ-0101216/pancreatic cancer/gemcitabine/proliferation分类
医药卫生引用本文复制引用
刘海潮,刘少朋,闫宏宪,白明辉,张继翔,李迎博,王闯,邹凯..下调Hsa-circ-0101216表达对胰腺癌吉西他滨化疗耐药的影响及其机制[J].解放军医学杂志,2025,50(6):656-664,9.基金项目
This work was supported by the Henan Province Medical Science and Technology Joint Construction Project(LHGJ20240736),the Guiding Science and Technology Plan Project of Luoyang Science and Technology Bureau(2302039Y),and the Henan Province Medical Science and Technology Key Project(SBGJ202102216) 河南省医学科技攻关联合共建项目(LHGJ20240736) （LHGJ20240736）
洛阳市科技局指导性科技计划项目(2302039Y) （2302039Y）
河南省医学科技攻关省部共建重点项目(SBGJ202102216) （SBGJ202102216）
