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目的:探讨人远端上游元件结合蛋白1(far upstream element binding protein 1,FUBP1)基因在肺腺癌组织中的表达及其对肺腺癌A549细胞化疗敏感性的影响。方法:选取43例来源于201-03-17-2012-05-09沈阳军区总医院胸外科,手术患者的肺腺癌及相应距肿瘤>1cm的癌旁组织标本为研究对象。Real-time PCR检测FUBP1mRNA在肺腺癌组织中的表达;RNA干扰技术沉默A549细胞中FUBP1基因的表达,蛋白质印迹法检测抑制效果;MTT法和流式细胞术测定FUBP1基因表达沉默对细胞生长抑制率和凋亡率的影响。不同浓度顺铂(DDP)处理FUBP1基因表达沉默的A549细胞,检测细胞生长抑制率和凋亡率。结果:FUBP1基因在肺腺癌组织中的表达显著上调,为癌旁正常组织中FUBP1的177.65%,t=2.858,P=0.006。FUBP1随着分化程度的降低和分期的升高呈逐渐增高趋势,且与淋巴结及远处转移相关,P<0.01。siRNA-FUBP1能够显著抑制A549细胞中FUBP1蛋白的表达,F=14.726,P<0.001;而FUBP1基因表达沉默的A549细胞的生长抑制率由1.4%升高至29.5%,F=16.302,P<0.001;凋亡率由2.68%升高至5.84%,F=19.497,P<0.001。FUBP1基因表达沉默的A549细胞分别经1、3和5μg/mL DDP处理后,生长抑制率由14.418%、17.460%和23.545%分别升高至21.693%、27.513%和37.566%,相同DDP浓度的2组间比较的t值分别为21.074、25.835和29.473,P值均<0.001;DDP的IC50由(5.12±0.31)μg/mL降至(3.93±0.24)μg/mL,t=3.487,P=0.001;而凋亡率由8.85%、14.37%和18.21%分别升高至13.25%、18.46%和26.52%,相同DDP浓度组间比较的t值分别为7.279、15.878和23.111,P值均<0.001。结论:FUBP1的表达上调与肺腺癌相关,FUBP1表达沉默能够增加A549细胞的生长抑制率和凋亡率,并且可以提高A549细胞对化疗药物DDP的敏感性。
Objective: To investigate the expression of human distal upstream element binding protein 1 (FUBP1) gene in lung adenocarcinoma and its effect on chemosensitivity of lung adenocarcinoma A549 cells. Methods: Forty-three cases of lung adenocarcinoma and corresponding para-cancerous tissue specimens> 1cm in diameter were selected from the Department of Thoracic Surgery, Surgery, General Hospital of Shenyang Military Command, 201-03-17-2012-05-09. Real-time PCR was used to detect the expression of FUBP1mRNA in lung adenocarcinoma; RNA interference technique was used to silence the expression of FUBP1 in A549 cells; Western blotting was used to detect the inhibitory effect; MTT assay and flow cytometry were used to determine the inhibitory effect of FUBP1 gene silencing on cell growth inhibition Effect of rate and apoptosis rate. A549 cells with silencing of FUBP1 gene were treated with different concentrations of cisplatin (DDP), and the cell growth inhibition rate and apoptosis rate were measured. Results: The expression of FUBP1 in lung adenocarcinoma was significantly up-regulated, which was 177.65% of FUBP1 in normal tissues adjacent to the tumor, t = 2.858, P = 0.006. FUBP1 gradually increased with the decrease of differentiation and staging, and was correlated with lymph node and distant metastasis, P <0.01. siRNA-FUBP1 could significantly inhibit the expression of FUBP1 protein in A549 cells, F = 14.726, P <0.001; while the inhibition rate of FUBP1 gene silencing A549 cells increased from 1.4% to 29.5%, F = 16.302, P <0.001 ; The apoptosis rate increased from 2.68% to 5.84%, F = 19.497, P <0.001. The growth inhibition rates of A549 cells with FUBP1 silencing A549 cells were increased from 14.418%, 17.460% and 23.545% to 21.693%, 27.513% and 37.566% respectively after being treated with 1, 3 and 5 μg / mL DDP, respectively The t values of the two groups were 21.074, 25.835 and 29.473, respectively, P <0.001. The IC50 of DDP decreased from 5.12 ± 0.31 μg / mL to 3.93 ± 0.24 μg / mL, t = 3.487, P = 0.001 While the apoptotic rates increased from 8.85%, 14.37% and 18.21% to 13.25%, 18.46% and 26.52%, respectively. The t values of the same DDP concentration group were 7.279, 15.878 and 23.111, respectively, P <0.001. Conclusion: Up-regulation of FUBP1 is associated with lung adenocarcinoma. Silencing of FUBP1 can increase the growth inhibition rate and apoptosis rate of A549 cells and increase the sensitivity of A549 cells to chemotherapeutic drug DDP.