CAJ | 학술논문

目的观察Toll样受体3(TLR3)对人肺腺癌细胞A549的增殖、抗凋亡及免疫逃逸方面的影响.方法给予TLR3-RNA干扰(RNAi)慢病毒转染组、未转染组和空载对照组TLR3的配体Poly(I∶C)刺激后,分别通过流式细胞术检测B7-H1的表达,实时定量聚合酶链反应(Real-time PCR)检测前列腺素E2(PGE-2)的表达,Western blot检测磷酸化p38的含量,并采用膜联蛋白V/碘化丙锭(Annexin V/PI)双染色法检测A549细胞的凋亡.结果在Poly(I∶C)刺激时间及浓度相同的情况下,转染TLR3-RNAi慢病毒载体的A549细胞组中B7-H1阳性细胞百分比、PGE-2的表达以及磷酸化p38的百分比相对值均低于未转染组和空载对照组(P＜0.01).凋亡实验显示,转染TLR3-RNAi慢病毒载体的A549细胞组细胞凋亡率[(44.40 ±1.41)％]明显高于未转染慢病毒载体组(34.70±0.89)％]和空载对照组[(36.50±1.12)％,P＜0.01.结论 TLR3基因沉默抑制了A549细胞中p38、PGE-2、B7-H1的表达,并降低了细胞的抗凋亡能力.
목적 관찰Toll양수체3(TLR3)대인폐선암세포A549적증식、항조망급면역도일방면적영향.방법 급여TLR3-RNA간우(RNAi)만병독전염조、미전염조화공재대조조TLR3적배체Poly(I∶C)자격후,분별통과류식세포술검측B7-H1적표체,실시정량취합매련반응(Real-time PCR)검측전렬선소E2(PGE-2)적표체,Western blot검측린산화p38적함량,병채용막련단백V/전화병정(Annexin V/PI)쌍염색법검측A549세포적조망.결과 재Poly(I∶C)자격시간급농도상동적정황하,전염TLR3-RNAi만병독재체적A549세포조중B7-H1양성세포백분비、PGE-2적표체이급린산화p38적백분비상대치균저우미전염조화공재대조조(P＜0.01).조망실험현시,전염TLR3-RNAi만병독재체적A549세포조세포조망솔[(44.40 ±1.41)％]명현고우미전염만병독재체조(34.70±0.89)％]화공재대조조[(36.50±1.12)％,P＜0.01.결론 TLR3기인침묵억제료A549세포중p38、PGE-2、B7-H1적표체,병강저료세포적항조망능력.
Objective Toll-like receptors (TLR) are very important innate immunity molecules.Recently the relationship between TLR and human tumors had been gradually focused on by more and more researchers.This study was to investigate the effect of TLR3 on the cell proliferation,anti-apoptosis and immune escape in human pulmonary adenocarcinoma cells.Methods A549 cells were transfected with TLR3-RNA interference (RNAi) lentiviral vector,blank vector or non-transfected,then incubated with TLR3 ligand Poly (I∶ C).The expression of B7-H1,prostaglandin E synthase-2 (PGE-2) and phosphorylated p38 were respectively evaluated by flow cytometry,real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting.Cells apoptosis was detected by flow cytometry with Annexin V fluoresceine isothiocyanate (FITC)/propidium iodide (PI) double staining.Results Under the same time and concentration stimulation with Poly (I∶ C),the percentage of B7-H1-positive cells,the expression of PGE-2 mRNA and the relative percentage of phosphoryated p38 in TLR3-RNAi lentiviral vector transfected A549 cells were significantly lower than those in the non-transfected and blank vector transfected A549 cells (P ＜ 0.01).The percentage of the apoptotic cells in TLR3-RNAi lentiviral vector transfected A549 cells [(44.40 ± 1.41) ％] was significantly higher than that in the non-transfected [(34.70 ± 0.89) ％] and blank vector transfected [(36.50 ± 1.12) ％] A549 cells (P ＜ 0.01).Conclusion Down-regulation of TLR3 could inhibit the expression of B7-H1,PGE-2 and phosphorylated p38 in A549 cells,and weaken the anti-apoptosis ability of A549 cells.