中华临床医师杂志(电子版)
中華臨床醫師雜誌(電子版)
중화림상의사잡지(전자판)
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2013年
22期
10107-10111
,共5页
白细胞介素6%滋养细胞%多耐药蛋白
白細胞介素6%滋養細胞%多耐藥蛋白
백세포개소6%자양세포%다내약단백
Interleukin-6%BeWo cells%Multidrug resistance protein 1
目的:探讨人滋养细胞多耐药蛋白(MDR1)mRNA 和蛋白表达与白细胞介素-6(IL-6)的相关性。方法体外培养人胎盘滋养细胞BeWo,分为IL-6干预组和空白对照组,每组根据干预时间不同又分为12 h、24 h、48 h和72 h组,每组重复三次。采用荧光定量PCR检测两组细胞MDR1 mRNA表达。蛋白免疫印迹法检测两组细胞MDR1蛋白表达。结果 RT-PCR检测干预12 h、24 h、48 h和72 h MDR1 mRNA表达量:IL-6干预组(1.04±0.11)×106 copies/μg RNA,(1.31±0.14)×106 copies/μg RNA,(1.68±0.13)×106 copies/μg RNA,(1.91±0.17)×106copies/μg RNA;空白对照组(1.00±0.09)×106 copies/μg RNA,(1.01±0.12)×106 copies/μg RNA,(1.00±0.11)×106 copies/μg RNA,(1.05±0.14)×106 copies/μg RNA。IL-6干预组MDR1 mRNA表达量显著高于相应时间点空白对照组的表达(F=3174.127,P=0.000)。IL-6处理组后一时间点的表达量显著高于前一时间点的表达量(F=574.180,P=0.000)。蛋白免疫印迹法检测干预12 h、24 h、48 h和72 h MDR1蛋白表达量:IL-6干预组0.08±0.01,0.11±0.01,0.13±0.02,0.15±0.01;空白对照组0.05±0.01,0.04±0.02,0.05±0.02,0.05±0.01。IL-6干预组MDR1蛋白表达量显著高于相应时间点空白对照组的表达(F=588.000,P=0.002)。IL-6处理组后一时间点的表达量显著高于前一时间点的表达量(F=26.000,P=0.001)。结论 IL-6能增强BeWo细胞MDR1 mRNA和蛋白的表达。
目的:探討人滋養細胞多耐藥蛋白(MDR1)mRNA 和蛋白錶達與白細胞介素-6(IL-6)的相關性。方法體外培養人胎盤滋養細胞BeWo,分為IL-6榦預組和空白對照組,每組根據榦預時間不同又分為12 h、24 h、48 h和72 h組,每組重複三次。採用熒光定量PCR檢測兩組細胞MDR1 mRNA錶達。蛋白免疫印跡法檢測兩組細胞MDR1蛋白錶達。結果 RT-PCR檢測榦預12 h、24 h、48 h和72 h MDR1 mRNA錶達量:IL-6榦預組(1.04±0.11)×106 copies/μg RNA,(1.31±0.14)×106 copies/μg RNA,(1.68±0.13)×106 copies/μg RNA,(1.91±0.17)×106copies/μg RNA;空白對照組(1.00±0.09)×106 copies/μg RNA,(1.01±0.12)×106 copies/μg RNA,(1.00±0.11)×106 copies/μg RNA,(1.05±0.14)×106 copies/μg RNA。IL-6榦預組MDR1 mRNA錶達量顯著高于相應時間點空白對照組的錶達(F=3174.127,P=0.000)。IL-6處理組後一時間點的錶達量顯著高于前一時間點的錶達量(F=574.180,P=0.000)。蛋白免疫印跡法檢測榦預12 h、24 h、48 h和72 h MDR1蛋白錶達量:IL-6榦預組0.08±0.01,0.11±0.01,0.13±0.02,0.15±0.01;空白對照組0.05±0.01,0.04±0.02,0.05±0.02,0.05±0.01。IL-6榦預組MDR1蛋白錶達量顯著高于相應時間點空白對照組的錶達(F=588.000,P=0.002)。IL-6處理組後一時間點的錶達量顯著高于前一時間點的錶達量(F=26.000,P=0.001)。結論 IL-6能增彊BeWo細胞MDR1 mRNA和蛋白的錶達。
목적:탐토인자양세포다내약단백(MDR1)mRNA 화단백표체여백세포개소-6(IL-6)적상관성。방법체외배양인태반자양세포BeWo,분위IL-6간예조화공백대조조,매조근거간예시간불동우분위12 h、24 h、48 h화72 h조,매조중복삼차。채용형광정량PCR검측량조세포MDR1 mRNA표체。단백면역인적법검측량조세포MDR1단백표체。결과 RT-PCR검측간예12 h、24 h、48 h화72 h MDR1 mRNA표체량:IL-6간예조(1.04±0.11)×106 copies/μg RNA,(1.31±0.14)×106 copies/μg RNA,(1.68±0.13)×106 copies/μg RNA,(1.91±0.17)×106copies/μg RNA;공백대조조(1.00±0.09)×106 copies/μg RNA,(1.01±0.12)×106 copies/μg RNA,(1.00±0.11)×106 copies/μg RNA,(1.05±0.14)×106 copies/μg RNA。IL-6간예조MDR1 mRNA표체량현저고우상응시간점공백대조조적표체(F=3174.127,P=0.000)。IL-6처리조후일시간점적표체량현저고우전일시간점적표체량(F=574.180,P=0.000)。단백면역인적법검측간예12 h、24 h、48 h화72 h MDR1단백표체량:IL-6간예조0.08±0.01,0.11±0.01,0.13±0.02,0.15±0.01;공백대조조0.05±0.01,0.04±0.02,0.05±0.02,0.05±0.01。IL-6간예조MDR1단백표체량현저고우상응시간점공백대조조적표체(F=588.000,P=0.002)。IL-6처리조후일시간점적표체량현저고우전일시간점적표체량(F=26.000,P=0.001)。결론 IL-6능증강BeWo세포MDR1 mRNA화단백적표체。
Objective To determine the relation between the level of IL-6 and the expression of MDR1 in BeWo cells. Methods In vitro cultured BeWo cells were treated with or without IL-6 for different time periods(12 h, 24 h, 48 h, 72 h), then the MDR1 mRNA transcripts were quantified by real-time PCR using relative determinations. MDR1 protein level were detected used western-blot. Results The expression of MDR1 mRNA of IL-6 groups were (1.04±0.11), (1.31±0.14),(1.68±0.13), (1.91±0.17)×106 copies/μg RNA, and the control groups were (1.00±0.09), (1.01±0.12), (1.00±0.11), (1.05±0.14)×106 copies/μg RNA, after BeWo cells were treated with or without IL-6 for 12 h, 24 h, 48 h, 72 h. Expression of MDR1 mRNA in each group of IL-6 groups was higher than that of the control groups(F=3174.127, P=0.000). And the expression of MDR1 mRNA increase remarkably with correspondence to time(F=574.180, P=0.000).The expression of MDR1 protein of IL-6 groups were 0.08±0.01, 0.11±0.01, 0.13±0.02, 0.15±0.01, and the control groups were 0.05±0.01, 0.04±0.02, 0.05 ±0.02, 0.05±0.01, after BeWo cells were treated with or without IL-6 for 12 h, 24 h, 48 h, 72 h. Western blot showed significant(F=588.000, P=0.000) increase in expression of MDR1 protein level in BeWo cells treated with IL-6 at 12 h, 24 h, 48 h and 72 h later. And the MDR1 protein level increase remarkably with correspondence to time(F=26.000, P=0.000). Conclusion MDR1 mRNA transcription and MDR1 protein level are significantly increased in BeWo cells after treated with IL-6.