乳腺癌耐药蛋白基因在肝癌组织的表达的临床意义及抑制其表达对肝癌细胞增殖凋亡的影响

The clinical significance of the expression of breast cancer resistance protein gene in hepatocellular carcinoma tissue and the effect of inhibiting its expression on the proliferation and apoptosis ofhepatocellular carcinoma cells

目的观察乳腺癌耐药蛋白（BCRP）在肝癌组织的表达与临床病理特征的关系及其表达的抑制对肝癌细胞增殖凋亡的影响。 方法通过反转录-聚合酶链反应（RT-PCR）检测肝癌组织中BCRP表达，并分析其与临床病理特征的关系；通过Lipofectamine？2000脂质体介导法将BCRP-小干扰RNA（siRNA）转染人肝癌SMMC-7721细胞，通过噻唑蓝（MTT）法、流式细胞仪及Western blot分别检测转染48 h后细胞的增殖、凋亡及BCRP、细胞核增殖抗原（Ki-67）、B细胞淋巴瘤/白血病-2相关X蛋白（bax）、Notch1、Hes1的蛋白表达。 结果BCRP在肝癌组织中的表达与肝癌患者的性别、年龄、肿瘤大小无明显相关，与病理分级、临床分期及肿瘤是否发生转移明显相关；BCRP-siRNA转染后SMMC-7721细胞中BCRP在转录及翻译水平的表达均受到抑制；BCRP-siRNA组细胞增殖（0.287±0.022）及Ki-67（0.142±0.018）、Notch1（0.162±0.021）、Hes1（0.111±0.015）蛋白表达均显著低于Control组（0.411±0.026、0.258±0.026、0.462±0.032、0.241±0.023；P增殖＝0.001，PKi-67＝0.001，PNotch1＝0.000，PHes1＝0.001），细胞凋亡率[（14.63±0.88）%]及bax蛋白表达（0.324±0.034）均显著高于Control组[（1.32±0.15）%，0.106±0.012；P凋亡＝0.000，Pbax＝0.000]。 结论肝癌中BCRP的表达与病理分级、临床分期及肿瘤是否发生转移相关，抑制其表达可降低肝癌细胞的增殖及诱导细胞的凋亡，其机制与调节Ki-67、bax表达及Notch1信号通路有关。

ObjectiveTo investigate the relationship between the expression of breast cancer resistance protein （BCRP） and the clinicopathological features in hepatocellular carcinoma （HCC） and the influence of the BCRP expression on the proliferation and apoptosis of HCC cells. MethodsTo detect the expression of BCRP in hepatocellular carcinoma tissues by reverse transcriptase-polymerase chain reaction （RT-PCR）, and analyze the relationship with clinicopathological parameters; BCRP-small interfering RNA （siRNA） was transfected into human hepatoma SMMC-7721 cells by Lipofectamine？ 2000 liposome mediated method; cell proliferation, apoptosis and BCRP, proliferation cell nuclear antigen （Ki-67）, B cell lymphoma/leukemia-2 associated X protein （bax）, Notch1 and Hes1 protein expression were detected by methyl thiazol tetrazolium （MTT） method, flow cytometry and Western blotting, respectively. ResultsThe expression of BCRP in HCC was not related to the sex, age and tumor size of HCC patients, and it was related to pathological grade, clinical stage and metastasis of tumor; expression of BCRP in SMMC-7721 cells after BCRP-siRNA was transfected cells was inhibited in the level of transcription and translation; cell proliferation （0.287±0.022） and expression of Ki-67 （0.142±0.018）, Notch1 （0.162±0.021） and Hes1 （0.111±0.015） protein in BCRP-siRNA group were significantly lower than Control group （0.411±0.026, 0.258±0.026, 0.462±0.032, 0.241±0.023; Pproliferation=0.001, PKi-67=0.001, PNotch1=0.000, PHes1=0.001）, the apoptosis rate （14.63±0.88）% and the expression of bax protein （0.324±0.034） was significantly higher than Control group （[1.32±0.15）%, 0.106±0.012; Papoptosis=0.000, Pbax=0.000]. ConclusionThe expression of BCRP is related to pathological grade, clinical stage and metastasis of HCC, inhibition of its expression can reduce the proliferation of hepatocellular carcinoma cells and induce cell apoptosis, the mechanism is related to the regulation of Ki-67, bax expression and Notch1 signaling pathway.