【摘要】 目的 探究镧离子(La³⁺)与酸根离子(SO₄^2-,Cl^-和NO₃^-)在镧盐诱导Hep G2细胞特定基因表达改变中的作用。方法 分别将含有La₂（SO₄）₃0.71 mmol·L^-1、La Cl₃1.41 mmol·L^-1、La（NO₃）₃1.41 mmol·L^-1、H₂SO₄2.12 mmol·L^-1、HCl 4.23 mmol·L^-1和HNO₃4.23 mmol·L^-1的培养液在CO₂培养箱中进行酸碱平衡1 h,p H值可恢复到7.25，随后分别处理对数生长期Hep G2细胞48 h。采用CCK-8试剂盒检测受试物对细胞存活率的影响。采用实时荧光定量PCR（RT-q PCR）检测丙氨酰氨基肽酶（ANPEP）、细胞色素P450家族1A1（CYP1A1）、氧化固醇结合蛋白样7（OSBPL7）、CYP3A5、钙电压门控通道辅助亚单位γ4（CACNG4）、双特异性磷酸酶4（DUSP4）、Ras蛋白特异性鸟嘌呤核苷酸释放因子1（RASGRF1）和CYP17A1 m RNA表达水平。结果 CCK-8法检测结果显示，与细胞对照组比较，3种镧盐和HCl处理组细胞存活率均降低（P<0.01），而H₂SO₄和HNO₃处理组细胞存活率均未见明显改变。RT-q PCR结果显示，与细胞对照组比较，La₂（SO₄）₃,La Cl₃和La（NO₃）₃均可诱导Hep G2细胞ANPEP,CYP1A1,OSBPL7,CYP3A5,CACNG4,DUSP4,RASGRF1和CYP17A1等8个基因m RNA表达上调（P<0.05）;SO₄^2-诱导ANPEP和CYP1A1表达（P<0.05）;Cl^-诱导CYP1A1表达（P<0.05）;NO₃^-诱导ANPEP和CYP3A5 m RNA表达，而抑制CYP1A1和CACNG4 m RNA表达（P<0.05）。归因分析发现，La₂（SO₄）₃和La Cl₃的La³⁺均能诱导Hep G2细胞上述8个基因m RNA表达上调（P<0.05），但对CYP1A1,CYP3A5,DUSP4和CYP17A1m RNA表达的诱导作用低于La₂（SO₄）₃（P<0.05），对CYP3A5和RASGRF1 m RNA表达的诱导作用低于La Cl₃（P<0.05）;La（NO₃）₃的La³⁺虽能诱导上述8个基因m RNA表达上调，但ANPEP m RNA的表达上调无统计学意义，且对CYP3A5和CYP17A1 m RNA表达的诱导作用低于La（NO₃）₃（P<0.05）。此外，3种镧盐的La³⁺对ANPEP,OSBPL7,CYP3A5,CACNG4,DUSP4,RASGRF1和CYP17A1等7个基因m RNA表达的诱导作用彼此间亦存在差异（P<0.05）。结论 La³⁺与酸根离子SO₄^2-,Cl^-和NO₃^-均可诱导Hep G2细胞特定基因m RNA表达的改变，La₂（SO₄）₃,La Cl₃和La（NO₃）₃对Hep G2细胞基因m RNA表达的影响是La³⁺与酸根离子SO₄^2-,Cl^-和NO₃^-共同作用的结果，不应仅归因于La³⁺。更多还原

【Abstract】 OBJECTIVE To explore the roles of lanthanum ions (La³⁺) and acid radical ions (SO₄^2-,Cl^-and NO₃^-) in the expression changes of specific genes in Hep G2 cells induced by lanthanum salts,respectively.METHODS A culture medium containing 0.71 mmol·L^-1lanthanum sulfate﹝La₂（SO₄）₃﹞,1.41 mmol·L^-1lanthanum chloride （La Cl₃）,1.41 mmol·L^-1lanthanum nitrate﹝La（NO₃）₃﹞,2.12 mmol·L^-1H₂SO₄,4.23 mmol·L^-1HCl or 4.23 mmol·L^-1HNO₃was placed in a CO₂incubator for acid-base balance for 1 h,and the p H value was restored to 7.25.Hep G2 cells in the logarithmic growth phase were treated with a acid-base-balanced culture medium containing the test chemical for 48 h.The effect of test chemicals on cell proliferation was detected by Cell Counting Kit-8 （CCK-8） assay.The m RNA expression levels of the target genes[alanyl aminopeptidase （ANPEP）,cytochrome P450 family 1 subfamily A member 1 （CYP1A1）,oxysterol binding protein like 7 （OSBPL7）,cytochrome P450 family 3 subfamily A member 5 （CYP3A5）,calcium voltage-gated channel auxiliary subunit gamma 4 （CACNG4）,dual specificity phosphatase 4 （DUSP4）,Ras protein specific guanine nucleotide releasing factor 1 （RASGRF1）and cytochrome P450 family 17 subfamily A member 1 （CYP17A1）]were determined by real-time quantitative PCR （RT-q PCR）.RESULTS CCK-8 assay results showed that compared with the cell control group,the relative proliferation rate （RPR） of cells decreased in lanthanum salt-treated and HCl-treated groups （P<0.01）,but remained unchanged in the H₂SO₄-treated or HNO₃-treated groups.RT-q PCR results showed that compared with the cell control group,La₂（SO₄）₃,La Cl₃and La（NO₃）₃all induced up-regulation of 8 genes detected in Hep G2 cells （P<0.05）,which were ANPEP,CYP1A1,OSBPL7,CYP3A5,CACNG4,DUSP4,RASGRF1 and CYP17A1,respectively.SO₄^2-induced up-regulation of ANPEP and CYP1A1 （P<0.05） while Cl^-induced up-regulation of CYP1A1 （P<0.05）.NO₃^-induced the expressions of ANPEP and CYP3A5,but repressed the expressions of CYP1A1 and CACNG4 （P<0.05）.Attribution analyses confirmed that both La₂（SO₄）₃-derived La³⁺and La Cl₃-derived La³⁺could induce the up-regulation of the above 8 genes in Hep G2 cells （P<0.05）,but the induction effect on CYP1A1,CYP3A5,DUSP4 and CYP17A1 was lower than that of La₂（SO₄）₃（P<0.05）,and the induction effect on CYP3A5 and RASGRF1 was lower than that of La Cl₃（P<0.05）.Although La（NO₃）₃-derived La³⁺could also induce the up-regulation of the above 8 genes,the up-regulation of ANPEP expression was not statistically significant,and the induction effect on CYP3A5 and CYP17A1 was lower than that of La（NO₃）₃（P<0.05）.In addition,there were also differences in the induction effects of La³⁺derived from the three lanthanum salts on the expressions of 7 genes including ANPEP,OSBPL7,CYP3A5,CACNG4,DUSP4,RASGRF1 and CYP17A1 （P<0.05）.CONCLUSION Both La³⁺and the acid radical ions SO₄^2-,Cl^-and NO₃^-can induce changes in the expressions of specific genes in Hep G2 cells.The effects of La₂（SO₄）₃,La Cl₃or La（NO₃）₃on gene expressions in Hep G2 cells result from the combined action of La³⁺and the acid radical ions SO₄^2-,Cl^-and NO₃^-,and should not be attributed to La³⁺alone.更多还原