CAJ | 학술논문

目的：探讨TRB3基因对人肺腺癌A549细胞生物学行为的影响。方法免疫组化及半定量RT-PCR测定NSCLC患者肿瘤组织中TRB3的表达，分析其与临床病理特征的相关性，构建shRNA TRB3重组质粒，脂质体法转染A549细胞，Transwell小室侵袭实验、RT-PCR、Western blot等方法探讨TRB3基因沉默后对A549细胞生物学行为的影响。结果<br>　　①在所有类型的NSCLC 中，尤其是腺癌中，TRB3表达上调。TRB3的表达与肿瘤大小、淋巴结转移、远处转移和复发相关，并与患者状况显著相关（P<0.05）。与自身癌旁组织相比，60例肺癌组织中TRB3 mRNA 水平明显增高（P<0.001）。此外， Kaplan-Meier 生存曲线表明TRB3表达与总体生存期和无病生存率呈负相关。②与对照组比较，shTRB3转染组TRB3表达明显降低。TRB3沉默后抑制A549细胞的生长。同时，Transwell 侵袭实验显示空载体转染组与对照组之间无显著性差异。与其他两组相比，shTRB3转染组侵袭率明显降低（P<0.001）。③与空白载体组相比，shTRB3 A549细胞中Notch1基因表达明显下调（P<0.001），两者呈正相关。结论沉默TRB3表达可抑制肺癌细胞生长，降低其侵袭能力，TRB3有可能成为治疗肺癌，预测其预后的一个新靶点。
목적：탐토TRB3기인대인폐선암A549세포생물학행위적영향。방법면역조화급반정량RT-PCR측정NSCLC환자종류조직중TRB3적표체，분석기여림상병리특정적상관성，구건shRNA TRB3중조질립，지질체법전염A549세포，Transwell소실침습실험、RT-PCR、Western blot등방법탐토TRB3기인침묵후대A549세포생물학행위적영향。결과<br>　　①재소유류형적NSCLC 중，우기시선암중，TRB3표체상조。TRB3적표체여종류대소、림파결전이、원처전이화복발상관，병여환자상황현저상관（P<0.05）。여자신암방조직상비，60례폐암조직중TRB3 mRNA 수평명현증고（P<0.001）。차외， Kaplan-Meier 생존곡선표명TRB3표체여총체생존기화무병생존솔정부상관。②여대조조비교，shTRB3전염조TRB3표체명현강저。TRB3침묵후억제A549세포적생장。동시，Transwell 침습실험현시공재체전염조여대조조지간무현저성차이。여기타량조상비，shTRB3전염조침습솔명현강저（P<0.001）。③여공백재체조상비，shTRB3 A549세포중Notch1기인표체명현하조（P<0.001），량자정정상관。결론침묵TRB3표체가억제폐암세포생장，강저기침습능력，TRB3유가능성위치료폐암，예측기예후적일개신파점。
Objective To investigate the influence of TRB3 on the biological behavior of human lung adenocarcinoma A549 cells. Methods The expression of TRB3 in NSCLC tissue was detected by immunochemistry and semi-quantitative RT-PCR, and its cor-relation with the clinical pathological features was analyzed. shRNA TRB3 recombinant plasmid was constructed, and transfected into A549 cells. The biological behavior of A549 cells was detected by Transwell chamber invasion assay, RT-PCR, Western blot, and so on. Results The expression of TRB3 was up-regulated in all types of NSCLC, especially in adenocarcinoma, but it is normal in normal lung tissue. It was related to tumor size, lymph node metastasis, distant metastasis, recurrence and the status of patients with NSCLC (P<0.05). Compared with the adjacent normal tissues, the level of TRB3 mRNA was significantly increased in the tissues of 60 cases with lung cancer (P<0.001). In addition, Kaplan-Meier survival curves showed that the expression of TRB3 was negatively correlated with the overall survival and disease-free survival (Fig. 1B, 1C).②Compared with control group, the expression of TRB3 was significantly de-creased in shTRB3 transfected group. The growth of A549 cells was inhibited after TRB3 knockdowned. At the same time, there was no significant difference between the empty vector transfected group and the control group. Compared with the other two groups, the inva-sion of the shTRB3 transfected group was significantly decreased (P<0.001).③Compared with the empty vector group, the expression of Notch1 in the shTRB3 transfected group was significantly down-regulated (P<0.001), and there was a positive correlation between them. Conclusion The decreased expression of TRB3 may inhibit the growth of A549 cells, and reduce its invasiveness. TRB3 would become a new target for treatment and prediction on the lung cancer prognosis.