PCB153对人肾上皮细胞中LINE-1表达及转座的影响
Effects of PCB153 on LINE-1 expression and translocation in human renal tubular epithelial cells
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摘要:
目的 探究2，2'，4，4'，5，5'-六氯联苯（PCB153）对人肾上皮细胞系（293T）细胞中长散在重复序列-1（LINE-1）基因表达和转座的影响。
方法 293T细胞每2 d传代一次，传代后立即使用5 μmol/L PCB153进行染毒。染毒后的细胞置于37℃、体积分数5% CO2培养箱中继续培养，连续染毒10 d。同时设置二甲基亚砜（DMSO）对照组。采用实时荧光定量PCR技术检测LINE-1开放阅读框1（ORF-1）和开放阅读框2（ORF-2）基因的mRNA表达水平及拷贝数，LINE-1拷贝数增加反映其发生转座。Western blot检测LINE-1 ORF-1和ORF-2蛋白表达水平。
结果 与对照组比较，从第4天开始PCB153染毒组细胞中LINE-1 ORF-1 mRNA表达水平增加（P < 0.05），从第6天开始PCB153染毒组细胞中LINE-1 ORF-2 mRNA表达水平增加（P < 0.05）。染毒第10天时，与对照组（LINE-1 ORF-1：2.74±0.51；LINE-1 ORF-2：0.57±0.10）相比，PCB153染毒组的LINE-1 ORF-1蛋白表达水平（4.39±0.85）和LINE-1 ORF-2蛋白表达水平（0.78±0.10）均明显增加（P < 0.05）。染毒第10天时，PCB153染毒组与对照组的LINE-1 ORF-1拷贝数分别为1.43±0.64和0.83±0.37，LINE-1 ORF-2拷贝数分别为1.11±0.52和0.81±0.12，差异均具有统计学意义（P < 0.05）。
结论 PCB153暴露可增加LINE-1的表达和拷贝数，诱导其转座。
Abstract:Objective To assess the effects of 2, 2', 4, 4', 5, 5'-hexachlorobiphenyl (PCB153) on long interspersed nuclear element-1 (LINE-1) gene expression and transposition in human renal tubular epithelial 293T cells.
Methods The 293T cells were transmitted every two days and exposed to PCB153 (5 μmol/L) in vitro after transmitting. The exposed cells were cultured in a 5% CO2 incubator at 37℃ and the exposure lasted for 10 days, with the 293T cells treated with dimethyl sulfoxide (DMSO) as control group. LINE-1 open read framework 1 (ORF-1) and open read framework 2 (ORF-2) gene mRNA expression levels and copy numbers (increasing copy number indicates transposition) were detected by real-time fluorescence quantitative PCR. LINE-1 ORF-1 and ORF-2 protein expression levels were measured by Western blot.
Results Compared with the control group, the mRNA expression level of LINE-1 ORF-1 in the PCB153 exposed group was increased from day 4 (P < 0.05), and the mRNA expression level of LINE-1 ORF-2 in the PCB153 exposed group was increased from day 6 (P < 0.05). On day 10, compared with the control group (LINE-1 ORF-1:2.74±0.51; LINE-1 ORF-2:0.57±0.10), the protein expression levels of LINE-1 ORF-1 (4.39±0.85) and LINE-1 ORF-2 (0.78±0.10) in the PCB153 exposed group were both increased (P < 0.05). In addition, on day 10, the copy numbers of LINE-1 ORF-1 were 1.43±0.64 and 0.83±0.37, and the copy numbers of LINE-1 ORF-2 were 1.11±0.52 and 0.81±0.12 in the PCB153 exposed group and the control group, respectively, with significant differences (P < 0.05).
Conclusion Exposure to PCB153 can increase the expression and copy number of LINE-1 and induce its transposition in 293T cells.
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