增殖1阻断基因在肝癌组织中上调并促进癌细胞的迁移

Block of proliferation 1 overexpression in hepatocellular carcinoma and its promoting effect on mi- gration of hepatocellular carcinoma HepG2 cells

目的观察增殖1阻断（BOPl）基因在肝细胞肝癌中的表达，以及对人肝癌细胞株HepG2增殖和迁移能力的影响。方法运用实时荧光定量聚合酶链反应（FQ—PCR）检测45例肝癌及正常组织中BOPlmRNA的表达量，比较其差异。构建pEGFP—N1．BOPl真核表达载体，并通过脂质体质粒转染HepG2细胞，运用细胞计数试剂盒（CCK_8）了解BOPl对细胞增殖的影响，划痕实验观察BOPl过表达对HepG2细胞运动迁移的影响。结果45例肝癌组织中BOPlmRNA表达水平ACt（为BOPl基因的ct值减去内参后所得）为15．164-1．86，对应正常组织表达水平△ct为17．48±2．68，癌组织中BOPl基因表达明显高于正常组织（P〈0．01）。细胞实验结果显示实验组吸光度值显著高于对照组（P〈0．05）；实验组向划痕部位的迁移速度明显快于对照组，72h对照组划痕部位仍有裂隙，而实验组完全愈合，差异有统计学意义（P〈0．05）。结论BOPl基因在肝癌组织中异常表达，癌组织中的表达明显高于正常组织；过表达BOPl可促进肝癌HepG2细胞的增殖和迁移能力。该基因可能在肝细胞肝癌的发生发展中起重要作用。

Objective To study the expression of block of proliferation 1 （ BOP1 ） gene in hepato- cellular carcinoma （HCC） and its impact on the proliferation and migration of HepG2 cell line. Methods Real-time fluorogenic quantitative polymerase chain reacton （RFQ-RCR） was conducted to quantify the ex- pression of BOP1 mRNA in the normal and cancerous tisseus from 45 patients with HCC. The eukaryotic vec- tor of human BOP1 was constructed, and transfected into HepG2 cells. The cell counting kit-8 （CCK-8） as- say and Scratch test were used to oberserve the influence of BOP1 overexpression on proliferation and mi- gration of HepG2 cells respectively. Results In 45 patients with HCC, the global expression （ △Ct） of BOP1 was （ 15.16±1.86） in tumor tissue and （ 17.48±2. 68） in corresponding normal tissue respective- ly. BOP1 mRNA expression in tumor tissues was significantly higher than in the normal tissues （P 〈 0.01 ）. The results of CCK-8 assay showed that the A value in experimental group was higher than in con- trol group, and overexpression of BOP1 significantly promoted cell proliferation （ P 〈0. 05 ）. Scratch text revealed that the speed of scratch healing in experimental group was significantly faster than in control group （P 〈 0.05）： Seventy-two h after transfection, the control group still had a fissure, but the experimental group had completely healed. Overexpression of BOP1 could obviously enhance the migration of HepG2 cells. Conclusion BOP1 gene has a higher expression level in HCC, and o,~erexpression of BOP1 can in- crease proliferatioh and migration of HepG2 cells. This gene probably plays an important role in carcinogen esis of HCC.