趋化因子受体CXCR7协同CXCR4调控胰腺癌趋向性转移及上皮间质转分化
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摘要
目的探究CXCL12/CXCR4/CXCR7生物轴在胰腺癌细胞侵袭转移及上皮间质转分化(EMT)中的调控作用及其作用机制,为胰腺癌转移的治疗研究提供新依据。方法利用CXCR4 shRNA及CXCR7 shRNA分别构建低表达CXCR4和CXCR7的胰腺癌细胞株。通过细胞侵袭和细胞迁移实验,观察CXCR4、CXCR7对CXCL12诱导的胰腺癌细胞侵袭转移的影响;Transwell侵袭实验及Western blot法检测间接共培养条件下胰腺癌细胞MiaPaCa-2的侵袭能力及EMT指标E-cadherin、Vimentin的变化。结果 CXCL12/CXCR4/CXCR7生物轴能够调控胰腺癌细胞的侵袭转移。CXCL12能显著增强胰腺癌细胞的侵袭转移能力,与单独沉默CXCR4或CXCR7相比,同时抑制CXCR4和CXCR7可以显著抑制CXCL12的促胰腺癌细胞侵袭转移作用。结论 CXCL12/CXCR4/CXCR7生物轴能够调控胰腺癌细胞侵袭转移及EMT发生。
Objective To investigate the effect of CXCL12/CXCR4/CXCR7 axis on the metastasis and invasion in pancreatic cancer so as to provide new evidence for research on pancreatic cancer metastasis treatment. Methods MiaPaCa-2 cells were transfected with CXCR4 shRNA and CXCR7 shRNA, and the Transwell assay was used to determine the effects of CXCL12/CXCR4/CXCR7 axis on cell invasion and migration. Quantitative RT-PCR and Western blotting were used to explore the effects of CXCL12/CXCR4/CXCR7 axis on the expressions of invasion-related genes(MMP-2 and uPA) and EMT-related genes(E-cadherin and Vimentin). Results CXCL12 significantly increased the metastasis and invasion of pancreatic cancer cells. The enhancement of tumor cell invasion was effectively countered by CXCR4 shRNA or CXCR7 shRNA. CXCL12/CXCR4 axis in cancer cells increased the expressions of invasion-related genes(MMP-2 and uPA) and EMT-related genes(E-cadherin and Vimentin). CXCL12/CXCR7 axis only increased the expressions of MMP-2 and uPA. Compared to blocking CXCR4 or CXCR7 alone, the inhibitory effects on invasion-related genes and EMT-related genes were more effective when both CXCR4 and CXCR7 were blocked. Conclusion CXCL12/CXCR4/CXCR7 axis regulates the EMT, metastasis, and invasion of pancreatic cancer cells.
Objective To investigate the effect of CXCL12/CXCR4/CXCR7 axis on the metastasis and invasion in pancreatic cancer so as to provide new evidence for research on pancreatic cancer metastasis treatment. Methods MiaPaCa-2 cells were transfected with CXCR4 shRNA and CXCR7 shRNA, and the Transwell assay was used to determine the effects of CXCL12/CXCR4/CXCR7 axis on cell invasion and migration. Quantitative RT-PCR and Western blotting were used to explore the effects of CXCL12/CXCR4/CXCR7 axis on the expressions of invasion-related genes(MMP-2 and uPA) and EMT-related genes(E-cadherin and Vimentin). Results CXCL12 significantly increased the metastasis and invasion of pancreatic cancer cells. The enhancement of tumor cell invasion was effectively countered by CXCR4 shRNA or CXCR7 shRNA. CXCL12/CXCR4 axis in cancer cells increased the expressions of invasion-related genes(MMP-2 and uPA) and EMT-related genes(E-cadherin and Vimentin). CXCL12/CXCR7 axis only increased the expressions of MMP-2 and uPA. Compared to blocking CXCR4 or CXCR7 alone, the inhibitory effects on invasion-related genes and EMT-related genes were more effective when both CXCR4 and CXCR7 were blocked. Conclusion CXCL12/CXCR4/CXCR7 axis regulates the EMT, metastasis, and invasion of pancreatic cancer cells.
引文
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[2] 牛坤汀,莫碧文,王志霞,等.CXCL12-CXCR4/CXCR7信号轴相关疾病研究进展[J].山东医药,2015,55(17):89-91.
[3] LEVOYE A,BALABANIAN K,BALEUX F,et al.CXCR7 heterodimerizes with CXCR4 and regulates CXCL12-mediated G protein signaling[J].Blood,2009,113(24):6085-6093.
[4] SOON LL.A discourse on cancer cell chemotaxis:Where to from here?[J].IUBMB Life,2007,59(2):60-67.
[5] KUMAR R,HERMAN JM,WOLFGANG CL,et al.Multidisciplinary management of pancreatic cancer[J].Surg Oncol Clin N Am,2013,22(2):265-287.
[6] DONà E,BARRY JD,VALENTIN G,et al.Directional tissue migration through a self-generated chemokine gradient[J].Nature,2013,503(7475):285-289.
[7] BURNS JM,SUMMERS BC,WANG Y,et al.A novel chemokine receptor for SDF-1 and I-TAC involved in cell survival,cell adhesion,and tumor development[J].J Exp Med,2006,203(9):2201-2213.
[8] HECKMANN D,MAIER P,LAUFS S,et al.The disparate twins:A comparative study of CXCR4 and CXCR7 in SDF-1alpha-induced gene expression,invasion and chemosensitivity of colon cancer[J].Clin Cancer Res,2014,20(3):604-616.
[9] XU Q,WANG Z,CHEN X,et al.Stromal-derived factor-1α/CXCL12-CXCR4 chemotactic pathway promotes perineural invasion in pancreatic cancer[J].Oncotarget,2015,6(7):4717-4732.
[10] LI X,MA Q,XU Q,et al.SDF-1/CXCR4 signaling induces pancreatic cancer cell invasion and epithelial-mesenchymal transition in vitro through non-canonical activation of Hedgehog pathway[J].Cancer Lett,2012,322(2):169-176.
[11] ELLENRIEDER V,ALBER B,LACHER U,et al.Role of MT-MMPs and MMP-2 in pancreatic cancer progression[J].Int J Cancer,2000,85(1):14-20.
[12] MAHADEVAN D,VON HOFF DD.Tumor-stroma interactions in pancreatic ductal adenocarcinoma[J].Mol Cancer Ther,2007,6(4):1186-1197.
[13] YILMAZ M,CHRISTOFORI G.EMT,the cytoskeleton,and cancer cell invasion[J].Cancer Metastasis Rev,2009,28(1-2):15-33.