CAJ | 학술논문

目的观察CXC趋化因子受体4(CXCR4)对人胆囊癌细胞株GBC-SD增殖的影响.方法分别用实时定量聚合酶链反应(Real-time PCR)、Western blot、流式细胞术等方法检测CXCR4在GBC-SD细胞及人脐静脉内皮细胞株ECV-304中mRNA、蛋白以及细胞表面分子的表达;设计CXCR4靶向短发卡RNA (shRNA),并构建到pSilencerTM2.1-U6质粒中,转染GBC-SD细胞,噻唑蓝比色法(MTT)检测GBC-SD细胞增殖.结果与ECV-304细胞比较,GBC-SD细胞中CXCR4 mRNA和蛋白呈明显阳性表达,而细胞表面CXCR4分子的表达也明显增高[(70.30±3.50)％与(0.76±0.11)％];与对照组GBC-SD细胞比较,转染了pSilencerTM 2.1-U6-CXCR4 shRNA质粒的GBC-SD细胞,其CXCR4 mRNA未见明显条带,蛋白表达亦明显降低,MTT结果显示干扰CXCR4 120 h后,GBC-SD细胞吸光度值较未转染组细胞明显降低(3.19±0.16与1.63±0.23).结论抑制CXCR4表达可抑制胆囊癌细胞生长.
목적 관찰CXC추화인자수체4(CXCR4)대인담낭암세포주GBC-SD증식적영향.방법 분별용실시정량취합매련반응(Real-time PCR)、Western blot、류식세포술등방법검측CXCR4재GBC-SD세포급인제정맥내피세포주ECV-304중mRNA、단백이급세포표면분자적표체;설계CXCR4파향단발잡RNA (shRNA),병구건도pSilencerTM2.1-U6질립중,전염GBC-SD세포,새서람비색법(MTT)검측GBC-SD세포증식.결과 여ECV-304세포비교,GBC-SD세포중CXCR4 mRNA화단백정명현양성표체,이세포표면CXCR4분자적표체야명현증고[(70.30±3.50)％여(0.76±0.11)％];여대조조GBC-SD세포비교,전염료pSilencerTM 2.1-U6-CXCR4 shRNA질립적GBC-SD세포,기CXCR4 mRNA미견명현조대,단백표체역명현강저,MTT결과현시간우CXCR4 120 h후,GBC-SD세포흡광도치교미전염조세포명현강저(3.19±0.16여1.63±0.23).결론 억제CXCR4표체가억제담낭암세포생장.
Objective To investigate the effect of CXC chemokine receptor 4 (CXCR4) on the growth of the gallbladder cancer cell line GBC-SD.Methods Using real-time quantitative polymerase chain reaction (Real-time PCR),Western blotting and flow cytometry,the expression of CXCR4 was detected in GBC-SD cells.After GBC-SD cells were transfected with CXCR4 shRNA,the proliferation was measured by MTT.Results The level of mRNA and protein of CXCR4 in GBC-SD cells,as well as the surface CXCR4 molecule on GBC-SD cells were all highly expressed than that in ECV-304 alls [(70.30 ±3.50)％ vs.(0.76 ± 0.11)％].The proliferation of GBC-SD cells was inhibited when CXCR4 was knocked down (3.19 ± 0.16 vs.1.63 ± 0.23).Conclusion The CXCR4 was highly expressed in GBC-SD cells,and the targeting of stromal cell derived factor-1 (SDF-1)/CXCR4 axis may be a novel strategy to control the progress of gallbladder cancer.