目的研究ITGA5经RNA干扰后对宫颈癌细胞生长和侵袭的影响。方法应用TNMplot、Kaplan-Meier Plotter、Human Protein Atlas数据库分析ITGA5在宫颈癌中的表达和对患者生存率的影响,采用两条靶向ITGA5的特异siRNA序列,进行RNA干扰实验。实...目的研究ITGA5经RNA干扰后对宫颈癌细胞生长和侵袭的影响。方法应用TNMplot、Kaplan-Meier Plotter、Human Protein Atlas数据库分析ITGA5在宫颈癌中的表达和对患者生存率的影响,采用两条靶向ITGA5的特异siRNA序列,进行RNA干扰实验。实验分为siITGA5#1干扰组、siITGA5#2干扰组和阴性对照组(非特异性siRNA)。实时定量PCR和Western blot法分别检测宫颈癌细胞中ITGA5的mRNA与蛋白水平,筛选内源性ITGA5呈高表达的HeLa及C-33A细胞做后续实验。Western blot法评估siRNA干扰后宫颈癌HeLa/C-33A细胞中ITGA5水平,并先后检测c-Myc、BCL-2、Bax及Vimentin、E-cadherin的表达变化,CCK-8法检测HeLa/C-33A细胞增殖,流式细胞术检测细胞凋亡,Transwell小室侵袭实验评估细胞侵袭能力。结果与阴性对照组相比,特异性siRNA显著下调宫颈癌HeLa/C-33A细胞中ITGA5蛋白的表达;宫颈癌细胞增殖明显受抑,细胞凋亡显著增加(P<0.05);c-Myc和BCL-2表达下降、Bax表达升高。干扰ITGA5后,HeLa/C-33A细胞侵袭能力显著降低,间质性标记物Vimentin的表达明显受抑,上皮细胞标记物E-cadherin表达水平明显增加。结论ITGA5经RNA干扰后,能明显抑制宫颈癌细胞生长和侵袭,ITGA5对宫颈癌细胞侵袭能力的影响可能是通过调控上皮细胞间质化来实现的。展开更多
The tumor–stroma interaction contributes to the aggressive and resistance nature of pancreatic ductal adenocarcinoma(PDAC),leading to treatment failure.Cancer-associated fibroblasts(CAFs),a key cell type in the strom...The tumor–stroma interaction contributes to the aggressive and resistance nature of pancreatic ductal adenocarcinoma(PDAC),leading to treatment failure.Cancer-associated fibroblasts(CAFs),a key cell type in the stroma,produce abundant extracellular matrix(ECM)and exhibit crosstalk with cancer cells inducing chemoresistance.In this study,we designed a cyclic peptide(cyAV3.3)targeting integrinα5(ITGA5)to disrupt CAF-induced desmoplasia and crosstalk with cancer cells.In vitro,cyAV3.3 inhibited the differentiation of pancreatic stellate cells into CAFs and reduced ECM production.In 3D co-cultured human spheroid models,the peptide decreased markers of resistance(ABCG1,BCL2,CXCR4),stemness(WNT1,CD44)and ECM remodeling(COL1A1,MMP2/9,LOX)and enhanced gemcitabine efficacy.In vivo,radiolabeled cyAV3.3 exhibited high tumor accumulation and retention following parenteral injections in a co-injection xenograft tumor model.Intriguingly,combination of cyAV3.3 with gemcitabine resulted in improved therapeutic efficacy of gemcitabine in co-injection xenograft and genetically engineered LSL-KrasG12D/+LSL-Trp53R172H/+Pdx1-Cre(KPC)PDAC models.These effects were attributed to reduced desmoplasia,vasculature compression and enhanced infiltration of cytotoxic T cells and apoptosis.This study presents a novel cyclic peptide inhibiting ITGA5-mediated tumor–stroma interaction and thereby reduce desmoplasia and resistance,ultimately enhancing chemotherapy efficacy in PDAC.展开更多
基金funded by Indonesia Endowment Fund for Education(Lembaga Pengelola Dana Pendidikan,to Deby Fajar Mardhian)ScarTec Therapeutics BV(Enschede,The Netherlands)+3 种基金Swedish Research Council(2011-5389 to Jai Prakash,The Netherlands)Dutch Organisation for Scientific Research(NWO-STW)Phase-2 grant(2017/STW/00364810 to Jai Prakash,The Netherlands)Egyptian Ministry of Higher Education and Scientific Research(20002907)Saadia A.Karim and Jennifer P.Morton were funded by a CRUK core program award to Jennifer P.Morton(A29996,UK).
文摘The tumor–stroma interaction contributes to the aggressive and resistance nature of pancreatic ductal adenocarcinoma(PDAC),leading to treatment failure.Cancer-associated fibroblasts(CAFs),a key cell type in the stroma,produce abundant extracellular matrix(ECM)and exhibit crosstalk with cancer cells inducing chemoresistance.In this study,we designed a cyclic peptide(cyAV3.3)targeting integrinα5(ITGA5)to disrupt CAF-induced desmoplasia and crosstalk with cancer cells.In vitro,cyAV3.3 inhibited the differentiation of pancreatic stellate cells into CAFs and reduced ECM production.In 3D co-cultured human spheroid models,the peptide decreased markers of resistance(ABCG1,BCL2,CXCR4),stemness(WNT1,CD44)and ECM remodeling(COL1A1,MMP2/9,LOX)and enhanced gemcitabine efficacy.In vivo,radiolabeled cyAV3.3 exhibited high tumor accumulation and retention following parenteral injections in a co-injection xenograft tumor model.Intriguingly,combination of cyAV3.3 with gemcitabine resulted in improved therapeutic efficacy of gemcitabine in co-injection xenograft and genetically engineered LSL-KrasG12D/+LSL-Trp53R172H/+Pdx1-Cre(KPC)PDAC models.These effects were attributed to reduced desmoplasia,vasculature compression and enhanced infiltration of cytotoxic T cells and apoptosis.This study presents a novel cyclic peptide inhibiting ITGA5-mediated tumor–stroma interaction and thereby reduce desmoplasia and resistance,ultimately enhancing chemotherapy efficacy in PDAC.
