胰腺导管腺癌中BCLAF1蛋白的表达

Expression of BCLAF1 protein in pancreatic ductal adenocarcinoma

目的探究Bcl-2相关转录因子-1(BCLAF1)蛋白在胰腺导管腺癌(PDAC)中的表达水平及临床意义。方法通过免疫组织化学法分别检测67例配对的PDAC以及癌旁组织石蜡切片中BCLAF1蛋白的表达。用实时荧光定量PCR(qRT-PCR)和Western blot检测8例配对的PDAC以及癌旁组织中BCLAF1蛋白的表达水平。PDAC Panc-1细胞中过表达BCLAF1后,通过Transwell侵袭实验观察PDAC细胞侵袭迁移能力的变化,采用CCK-8法检测各组PDAC细胞增殖情况。结果免疫组化结果显示:BCLAF1主要表达于PDAC导管细胞的细胞核中,PDAC组BCLAF1阳性表达的有54例(80.59%),高于癌旁组织(35.82%),差异有统计学意义(χ^2=27.610,P<0.001)。Western blot结果表明:PDAC中BCLAF1蛋白含量高于癌旁组织,差异有统计学意义(P<0.001)。PDAC细胞中过表达BCLAF1后明显增加胰腺细胞的侵袭迁移能力,同时能促进癌细胞的增殖能力。BCLAF1在PDAC癌组织中的表达水平与患者肿瘤的大小、TNM分期、有无淋巴结转移以及临床分期和预后情况等临床生物学特征显著相关。Kaplan-Meier生存分析显示BCLAF1高表达生存时间比BCLAF1低表达生存时间显著缩短,差异有统计学意义(χ^2=17.419,P<0.01)。COX回归多因素分析显示BCLAF1和TNM分期是影响PDAC预后的独立因素。结论 BCLAF1蛋白表达水平在PDAC中显著升高,转染pcDNA3.1-BCLAF-1质粒Panc-1细胞增殖速度,侵袭能力得到提高,其检测有助于对PDAC的早期诊断以及预后评估。

Objective To investigate the expression level and clinical significance of Bcl-2 associated transcription factor 1(BCLAF1) protein in pancreatic ductal adenocarcinoma(PDAC). Methods Immunohistochemistry was used to detect the expression of BCLAF1 protein in 67 pairs of PDAC and paraffin sections of paracancerous tissues. Real-time quantitative PCR(qRT-PCR) and Western blot were used to detect the expression levels of BCLAF1 protein in 8 matched PDACs and adjacent tissues. After overexpression of BCLAF1 in panc-1 cells of pancreatic cancer, the invasion and migration ability of pancreatic cancer cells were observed by Transwell invasion assay. The proliferation of pancreatic cancer cells in each group was detected by CCK-8 method.Results The results of immunohistochemistry showed that BCLAF1 was mainly expressed in the nucleus of PDAC catheter cells. 66 cases(80.59%) were positive for BCLAF1 in PDAC group, which was higher than that in adjacent tissues(35.82%).Statistical significance was significant(χ2= 27. 610,P < 0. 001). Western blot results showed that the content of BCLAF1 protein in PDAC was higher than that in adjacent tissues,and the difference was statistically significant( P < 0. 001). Overexpression of BCLAF1 in pancreatic cancer cells significantly increases the invasion and migration ability of pancreatic cells,and at the same time promotes the proliferation of cancer cells. The expression level of BCLAF1 in pancreatic ductal adenocarcinoma was significantly correlated with clinical tumor characteristics such as tumor size,TNM stage,lymph node metastasis,clinical stage and prognosis. Kaplan-Meier survival analysis showed that the high expression survival time of BCLAF1 was significantly lower than that of BCLAF1(χ2=17. 419,P < 0. 01). Multivariate analysis of COX regression showed that BCLAF1 and TNM staging were independent factors influencing the prognosis of pancreatic cancer. Conclusion The expression level of BCLAF1 protein is significantly increased in PDAC,and the proliferation rate of PANC-1 cells transfect with pcDNA3. 1-BCLAF-1 plasmid is improved. Its detection contributes to the early diagnosis and prognosis assessment of pancreatic ductal adenocarcinoma.