苯代谢物氢醌对K562细胞WRN基因转录及表达的影响
发布时间：2018-10-31 19:58
【摘要】：目的探讨苯代谢物氢醌(HQ)对人白血病细胞K562中解旋酶WRN基因mRNA转录及蛋白表达的影响。方法常规培养白血病细胞K562至生长对数期,低剂量HQ组、中剂量HQ组、高剂量HQ组分别以15、30、60μmol/L浓度HQ重复间隔染毒48及72 h,以等体积的PBS培养的细胞组为空白对照组。采用单细胞凝胶电泳法(SCGE)检测细胞DNA损伤;采用Taqman探针实时荧光定量PCR法检测WRN基因mRNA水平,采用免疫印迹分析WRN基因蛋白表达水平,计算mRNA及蛋白的相对表达量。结果 (1)HQ可导致细胞DNA损伤,且损伤效应随染毒浓度增加而增大,72 h比48 h的DNA损伤程度增加,呈时间-剂量依赖性;(2)HQ染毒48 h,WRN基因mRNA在各组差异均无统计学意义(P0.05);HQ染毒72 h,各剂量组与空白对照组比较差异有统计学意义(P0.05),低、中剂量组与高剂量组比较,差异有统计学意义(P0.05),低剂量组与中剂量组组间差异无统计学意义(P0.05);(3)HQ染毒48 h,WRN蛋白相对表达量在各组差异无统计学意义(P0.05);HQ染毒72 h,各剂量组与空白对照组比较,蛋白表达随着染毒剂量增大而降低,差异有统计学意义(P0.05),HQ各剂量组之间两两比较差异无统计学意义(P0.05)。结论 HQ可导致K562细胞DNA损伤,且呈时间-剂量依赖性,其机制可能与HQ下调WRN基因mRNA转录及蛋白的表达有关。
[Abstract]:Objective to investigate the effect of benzene metabolite hydroquinone (HQ) on mRNA transcription and protein expression of helicase WRN gene in human leukemia cell line K562. Methods K562 leukemic cells were cultured from normal culture to logarithmic phase, low dose HQ group, middle dose HQ group and high dose HQ group were exposed to HQ at 1530 ~ 60 渭 mol/L concentration for 48 h and 72 h, respectively. The cells cultured with the same volume of PBS were used as blank control group. Single cell gel electrophoresis (SCGE) was used to detect DNA damage, Taqman probe real-time fluorescence quantitative PCR method was used to detect the mRNA level of WRN gene, and Western blotting was used to analyze the expression level of WRN gene protein, and to calculate the relative expression of mRNA and protein. Results (1) HQ induced DNA damage in cells, and the damage effect increased with the increase of the concentration of DNA. The degree of DNA damage was increased at 72 h than 48 h in a time-dose dependent manner. (2) there was no significant difference in mRNA of WRN gene in 48 h after HQ exposure (P0.05); 72 hours after exposure to HQ, the difference between each dose group and the blank control group was statistically significant (P0.05), and the difference between the middle dose group and the high dose group was statistically significant (P0.05), the difference between the middle dose group and the high dose group was significant (P0.05). There was no significant difference between the low dose group and the middle dose group (P0.05). (3) there was no significant difference in the relative expression of WRN protein in each group at 48 h after exposure to HQ (P0.05); After 72 hours of HQ exposure, the protein expression decreased with the increase of the dose of HQ compared with the blank control group (P0.05). There was no significant difference between the two groups (P0.05). Conclusion HQ can induce DNA damage in K562 cells in a time-dose dependent manner, and its mechanism may be related to the down-regulation of WRN gene mRNA transcription and protein expression by HQ.
【作者单位】： 贵州医科大学医学检验学院;贵阳中医学院第二附属医院检验科;
【基金】：国家自然科学基金资助项目(编号:81360437)
【分类号】：R114
[Abstract]:Objective to investigate the effect of benzene metabolite hydroquinone (HQ) on mRNA transcription and protein expression of helicase WRN gene in human leukemia cell line K562. Methods K562 leukemic cells were cultured from normal culture to logarithmic phase, low dose HQ group, middle dose HQ group and high dose HQ group were exposed to HQ at 1530 ~ 60 渭 mol/L concentration for 48 h and 72 h, respectively. The cells cultured with the same volume of PBS were used as blank control group. Single cell gel electrophoresis (SCGE) was used to detect DNA damage, Taqman probe real-time fluorescence quantitative PCR method was used to detect the mRNA level of WRN gene, and Western blotting was used to analyze the expression level of WRN gene protein, and to calculate the relative expression of mRNA and protein. Results (1) HQ induced DNA damage in cells, and the damage effect increased with the increase of the concentration of DNA. The degree of DNA damage was increased at 72 h than 48 h in a time-dose dependent manner. (2) there was no significant difference in mRNA of WRN gene in 48 h after HQ exposure (P0.05); 72 hours after exposure to HQ, the difference between each dose group and the blank control group was statistically significant (P0.05), and the difference between the middle dose group and the high dose group was statistically significant (P0.05), the difference between the middle dose group and the high dose group was significant (P0.05). There was no significant difference between the low dose group and the middle dose group (P0.05). (3) there was no significant difference in the relative expression of WRN protein in each group at 48 h after exposure to HQ (P0.05); After 72 hours of HQ exposure, the protein expression decreased with the increase of the dose of HQ compared with the blank control group (P0.05). There was no significant difference between the two groups (P0.05). Conclusion HQ can induce DNA damage in K562 cells in a time-dose dependent manner, and its mechanism may be related to the down-regulation of WRN gene mRNA transcription and protein expression by HQ.
【作者单位】： 贵州医科大学医学检验学院;贵阳中医学院第二附属医院检验科;
【基金】：国家自然科学基金资助项目(编号:81360437)
【分类号】：R114
【相似文献】
相关期刊论文 前10条
1 郝坡;刘北忠;孟凡萍;欧阳峰;王东生;刘畅;钟梁;金丹婷;王，
本文编号：2303376
本文链接：https://www.wllwen.com/yixuelunwen/yufangyixuelunwen/2303376.html
