镉对肾小管上皮细胞中原癌基因c-myc表达的影响
|
摘要
目的探讨镉对肾小管上皮细胞(TEC)毒效应及原癌基因c-myc表达的影响。方法将处于对数生长期的TEC分别暴露于含终浓度为0(对照)、0.625、1.25、2.50、5.0、10.0μmol/L氯化镉的培养基染毒24、48、72 h后,采用MTT法测定TEC的存活情况。将处于对数生长期的TEC分别暴露于含终浓度为0(对照)、0.625、1.25、2.50μmol/L氯化镉的培养基染毒24、48、72 h后,采用RT-qPCR技术检测TEC c-myc mRNA的表达水平。结果与对照组比较,2.50、5.0μmol/L氯化镉染毒24、48、72 h的TEC存活率较高,除5.0μmol/L氯化镉染毒72 h外,差异均有统计学意义(P<0.05)。随着氯化镉染毒剂量的升高,不同染毒时间TEC的存活率均呈先上升后下降的趋势,且均以2.50μmol/L组最高;随着氯化镉染毒时间的延长,2.50、5.0、10.0μmol/L组TEC的存活率均呈下降趋势。与对照组比较,各剂量氯化镉染毒不同时间后TEC中c-myc mRNA的表达水平均较高,差异有统计学意义(P<0.05);且随着氯化镉染毒剂量的升高,不同染毒时间TEC中c-myc mRNA的表达水平均呈上升趋势。结论一定低剂量镉诱导TEC中c-myc持续激活和过表达与镉诱导TEC过度增殖密切相关,可能是镉致肾损伤的毒作用机制之一。
Objective To explore the toxic effects of cadmium on the renal tubular epithelial cell(TEC) and study the effects of cadmium on the expression of proto-oncogene c-myc in TEC. Methods The TEC in logarithmic growth phase were exposed to serum medium containing cadmium chloride at concentrations of 0,0.625,1.25,2.50,5.0,10.0 μmol/L,respectively. After exposure for 24,48,72 h,MTT method was used to detect the survival of TEC in each group. The TEC in logarithmic growth phase were exposed to serum medium containing cadmium chloride at concentrations of 0,0.625,1.25,2.50 μmol/L,respectively. After exposure for 24,48,72 h,RT-qPCR technique was used to observe mRNA relative expression level of c-myc gene. Results After exposure for 24,48,72 h,survival rates of TEC in 2.50 and 5.0 μmol/L exposure groups were high and showed a significant difference(P<0.05) compared with the control group, except 2.50 μmol/L exposure group for 72 h.The survival of TEC at different time increased first and then decreased with the increase of exposure dosage,meanwhile,the survival rate in 2.50 μmol/L exposure group were the highest among all groups. As the extension of exposure time,the survival rates of TEC in 2.50, 5.0 and 10.0 μmol/L exposure groups showed a downward trend. Compared with the control group, the expression level of c-myc gene in all exposure groups increased and the difference was significant(P<0.05). With the increase of exposure dosage,the expression level of c-myc gene at different exposure time showed an increasing trend. Conclusion The sustained activation and over-expression of c-myc in TEC induced by a certain dosage of cadmium is closely related with proliferation induced by cadmium,which maybe one of the mechanisms of renal damage induced by cadmium.
Objective To explore the toxic effects of cadmium on the renal tubular epithelial cell(TEC) and study the effects of cadmium on the expression of proto-oncogene c-myc in TEC. Methods The TEC in logarithmic growth phase were exposed to serum medium containing cadmium chloride at concentrations of 0,0.625,1.25,2.50,5.0,10.0 μmol/L,respectively. After exposure for 24,48,72 h,MTT method was used to detect the survival of TEC in each group. The TEC in logarithmic growth phase were exposed to serum medium containing cadmium chloride at concentrations of 0,0.625,1.25,2.50 μmol/L,respectively. After exposure for 24,48,72 h,RT-qPCR technique was used to observe mRNA relative expression level of c-myc gene. Results After exposure for 24,48,72 h,survival rates of TEC in 2.50 and 5.0 μmol/L exposure groups were high and showed a significant difference(P<0.05) compared with the control group, except 2.50 μmol/L exposure group for 72 h.The survival of TEC at different time increased first and then decreased with the increase of exposure dosage,meanwhile,the survival rate in 2.50 μmol/L exposure group were the highest among all groups. As the extension of exposure time,the survival rates of TEC in 2.50, 5.0 and 10.0 μmol/L exposure groups showed a downward trend. Compared with the control group, the expression level of c-myc gene in all exposure groups increased and the difference was significant(P<0.05). With the increase of exposure dosage,the expression level of c-myc gene at different exposure time showed an increasing trend. Conclusion The sustained activation and over-expression of c-myc in TEC induced by a certain dosage of cadmium is closely related with proliferation induced by cadmium,which maybe one of the mechanisms of renal damage induced by cadmium.
引文
[1]杜丽娜,余若祯,王海燕,等.重金属镉污染及其毒性研究进展[J].环境与健康杂志,2013,30(2):167-174.
[2]吴小胜,魏帅,魏益民,等.镉肾脏毒性生物标志物的研究进展[J].环境与健康杂志,2011,28(8):739-742.
[3]杨光,王少鹏,李振雪.镉所致肾毒性及氧化性损害机制的研究[J].毒理学杂志,2007,21(5):398-401.
[4]王玥.长期低剂量镉致正常肝脏细胞异常增殖及其与caspase-8甲基化的关系[D].吉林:吉林大学,2015.
[5]楼哲丰,曹琼洁,冯钰淇,等.镉对果蝇肠道上皮细胞损伤和调控中肠干细胞增殖、分化机制的研究[J].中国细胞生物学学报,2013,35(5):602-608.
[6]詹启敏,陈杰.细胞周期与肿瘤转化医学[J].中国肿瘤临床,2014,41(1):1-7.
[7]程倩,袁富文,童坦君.从分子水平看c-Myc在细胞衰老中的作用[J].生物化学与生物物理进展,2014,41(3):266-272.
[8]何锦园,曾国华,吴文起,等.SD大鼠肾小管上皮细胞的原代培养和鉴定[J].东南大学学报:医学版,2010,29(4):395-399.
[9]彭单单,程力平.肾小管上皮细胞损伤的研究进展[J].临床儿科杂志,2013,31(6):584-587.
[10]程叙扬,李晓玫,王海燕.肾小管上皮细胞损伤在慢性肾脏疾病进展中的作用[J].国外医学泌尿系统分册,2002,22(3):152-156.
[11]王心如.毒理学基础[M].6版.北京:人民卫生出版社,2012:438-442.
[12]戈果,周静,谢骐骥,等.氯化镉对大鼠原代星形胶质细胞凋亡的影响[J].环境与健康杂志,2015,32(2):102-104.
[13]吴晓丽,任香梅.镉诱导肾细胞凋亡及其对PI3K/Akt和ERK1/2信号通路的影响[J].热带医学杂志,2013,13(12):1452-1455.
[14]王海江,任来成.细胞、细胞因子与肾间质纤维化关系[J].山西医科大学学报,2007,38(3):266-269.
[15]宋新文,邓存良.细胞因子与肾小管间质纤维化[J].国外医学:泌尿系统分册,2005,25(3):374-377.
[16]邓景岳,毕明刚.影响原癌基因c-myc作用因素的研究进展[J].肿瘤,2009,29(12):1176-1179.
[17]白阳,叶健,王敬泽.c-myc功能及其下游靶点[J].细胞生物学杂志,2007,29(2):191-196.
[2]吴小胜,魏帅,魏益民,等.镉肾脏毒性生物标志物的研究进展[J].环境与健康杂志,2011,28(8):739-742.
[3]杨光,王少鹏,李振雪.镉所致肾毒性及氧化性损害机制的研究[J].毒理学杂志,2007,21(5):398-401.
[4]王玥.长期低剂量镉致正常肝脏细胞异常增殖及其与caspase-8甲基化的关系[D].吉林:吉林大学,2015.
[5]楼哲丰,曹琼洁,冯钰淇,等.镉对果蝇肠道上皮细胞损伤和调控中肠干细胞增殖、分化机制的研究[J].中国细胞生物学学报,2013,35(5):602-608.
[6]詹启敏,陈杰.细胞周期与肿瘤转化医学[J].中国肿瘤临床,2014,41(1):1-7.
[7]程倩,袁富文,童坦君.从分子水平看c-Myc在细胞衰老中的作用[J].生物化学与生物物理进展,2014,41(3):266-272.
[8]何锦园,曾国华,吴文起,等.SD大鼠肾小管上皮细胞的原代培养和鉴定[J].东南大学学报:医学版,2010,29(4):395-399.
[9]彭单单,程力平.肾小管上皮细胞损伤的研究进展[J].临床儿科杂志,2013,31(6):584-587.
[10]程叙扬,李晓玫,王海燕.肾小管上皮细胞损伤在慢性肾脏疾病进展中的作用[J].国外医学泌尿系统分册,2002,22(3):152-156.
[11]王心如.毒理学基础[M].6版.北京:人民卫生出版社,2012:438-442.
[12]戈果,周静,谢骐骥,等.氯化镉对大鼠原代星形胶质细胞凋亡的影响[J].环境与健康杂志,2015,32(2):102-104.
[13]吴晓丽,任香梅.镉诱导肾细胞凋亡及其对PI3K/Akt和ERK1/2信号通路的影响[J].热带医学杂志,2013,13(12):1452-1455.
[14]王海江,任来成.细胞、细胞因子与肾间质纤维化关系[J].山西医科大学学报,2007,38(3):266-269.
[15]宋新文,邓存良.细胞因子与肾小管间质纤维化[J].国外医学:泌尿系统分册,2005,25(3):374-377.
[16]邓景岳,毕明刚.影响原癌基因c-myc作用因素的研究进展[J].肿瘤,2009,29(12):1176-1179.
[17]白阳,叶健,王敬泽.c-myc功能及其下游靶点[J].细胞生物学杂志,2007,29(2):191-196.