蛋白酶体β亚基4型对人肝癌SMMC7721细胞增殖存活的影响  

Effect of proteasome subunit beta type 4 on the proliferation and viability of human liver cancer SMMC7721 cells

作　　者：陆谦[1,2] 程欣 夏振国 陈钟 LU Qian;CHENG Xin;XIA Zhenguo(Department of Hepatobiliary Surgery,Affiliated Hospital of Nantong University,Nantong,Jiangsu 226001,China)

机构地区：[1]南通大学附属医院肝胆外科,江苏南通226001 [2]南通市通州区人民医院普外科,江苏南通226300

出　　处：《临床肝胆病杂志》2020年第3期592-595,共4页Journal of Clinical Hepatology

基　　金：国家自然科学基金(81070360)。

摘　　要：目的旨在探讨蛋白酶体β亚基4型(PSMB4)对人肝癌SMMC7721细胞增殖存活的影响及其可能的机制。方法利用特异性短发夹RNA(shRNA)技术构建干扰PSMB4表达的SMMC7721实验组细胞;运用MTT、克隆形成实验检测细胞增殖的影响,流式细胞术检测细胞凋亡率的变化,Western Blot检测相关蛋白的表达变化。计量资料两组间比较采用独立样本t检验。结果成功构建了干扰PSMB4表达的SMMC7721实验组细胞(shRNA1:t=22.67,P<0.0001;shRNA2;t=30.88,P<0.0001;shRNA3:t=67.82,P<0.0001)。MTT实验显示第4天(0.4770±0.0135 vs 0.3237±0.0127,t=8.286,P=0.0012)、第5天(0.5893±0.0088 vs 0.3847±0.0090,t=16.220,P<0.0001)实验组细胞OD490值显著低于对照组。克隆形成实验显示细胞集落数显著减少。流式细胞术显示实验组细胞的早期凋亡率[(5.5570±0.2589)%vs(3.8870±0.3324)%,t=3.964,P=0.0166]、晚期凋亡率[(12.6300±0.4198)%vs(5.3100±0.3062)%,t=14.080,P=0.0001]及总凋亡率[(18.1800±0.6785)%vs(9.1970±0.6313)%,t=9.967,P=0.0006]均明显高于对照组,且干扰PSMB4表达的实验组细胞中核因子-κB亚基p65蛋白表达降低(0.8015±0.0120 vs 0.2841±0.0110,t=31.830,P<0.0001),核因子-κB抑制蛋白α表达增加(0.4816±0.0112 vs 0.6583±0.0142,t=9.774,P=0.0006)。结论干扰PSMB4表达可能通过抑制NF-κB信号通路,从而导致肝癌SMMC7721细胞的增殖存活能力下降。Objective To investigate the effect of proteasome subunit beta type 4(PSMB4)on the proliferation and viability of human liver cancer SMMC7721 cells and its possible mechanisms.Methods The specific short-hairpin RNA(shRNA)technique was used to construct SMMC7721 cells with knockdown expression of PSMB4,and these cells were selected as experimental group.MTT assay and colony-forming assay were used to observe the change in cell proliferation,flow cytometry was used to measure the change in cell apoptosis rate,and Western blot was used to measure the change in the expression of related proteins.The independent samples t-test was used for comparison of continuous data between two groups.Results SMMC7721 cells with knockdown expression of PSMB4 were successfully constructed(shRNA1:t=22.67,P<0.0001;shRNA2:t=30.88,P<0.0001;shRNA3:t=67.82,P<0.0001).The MTT assay showed that the experimental group had a significantly lower OD490 value than the control group on day 4(0.4770±0.0135 vs 0.3237±0.0127,t=8.286,P=0.0012)and day 5(0.5893±0.0088 vs 0.3847±0.0090,t=16.220,P<0.0001).The colony-forming assay showed a significant reduction in the number of cell colonies in the experimental group.Flow cytometry showed that compared with the control group,the experimental group had significantly higher early apoptosis rate(5.5570%±0.2589%vs 3.8870%±0.3324%,t=3.964,P=0.0166),late apoptosis rate(12.6300%±0.4198%vs 5.3100%±0.3062%,t=14.080,P=0.0001),and total apoptosis rate(18.1800%±0.6785%vs 9.1970%±0.6313%,t=9.967,P=0.0006),as well as a significant reduction in the protein expression of nuclear factor-kappa B p65(0.8015±0.0120 vs 0.2841±0.0110,t=31.830,P<0.0001)and a significant increase in the protein expression of nuclear factor-kappa B inhibitory proteinα(0.4816±0.0112 vs 0.6583±0.0142,t=9.774,P=0.0006).Conclusion Knockdown of PSMB4 expression may reduce the proliferation and viability of liver cancer SMMC7721 cells by inhibiting the NF-κB signaling pathway.

关 键 词：癌 肝细胞 蛋白酶体β亚基4型 细胞增殖 细胞存活 

分 类 号：R735.7[医药卫生—肿瘤]