国际生物医学工程杂志
國際生物醫學工程雜誌
국제생물의학공정잡지
INTERNATIONAL JOURNAL OF BIOMEDICAL ENGINEERING
2012年
2期
74-78
,共5页
徐淑霞%陈丹丹%马凤霞%及月茹%戎丽娟%陈芳%池颖%杨少光%卢士红%韩忠朝
徐淑霞%陳丹丹%馬鳳霞%及月茹%戎麗娟%陳芳%池穎%楊少光%盧士紅%韓忠朝
서숙하%진단단%마봉하%급월여%융려연%진방%지영%양소광%로사홍%한충조
MDA-MB-231%TLR3%Poly(I:C)%增殖%凋亡
MDA-MB-231%TLR3%Poly(I:C)%增殖%凋亡
MDA-MB-231%TLR3%Poly(I:C)%증식%조망
MDA-MB-231%TLR3%Poly(I:C)%Proliferation%Apoptosis
目的 Toll样受体(TLRs)在肿瘤的发生发展和肿瘤免疫中起到重要的作用,有报道称TLR3的激活能够促进某些肿瘤细胞的凋亡.本研究旨在探讨TLR3的特异性激动剂Poly(I:C)对恶性程度较强的乳腺癌细胞MDA-MB-231增殖与凋亡的影响,并初步探讨其发挥作用的机制.方法 反转录PCR检测Toll样受体各个亚型在mRNA水平上的表达；CCK-8细胞活力试剂盒检测不同浓度Poly(I:C)对MDA-MB-231细胞增殖的影响；流式细胞术用Annexin V-FITC/PI染色检测Poly(I:C)对MDA-MB-231细胞凋亡作用的影响；实时定量反转录PCR检测相关细胞因子的表达变化.结果 在mRNA水平上,MDA-MB-231细胞TLRs各亚型中TLR3表达最高；CCK-8检测结果显示Poly(I:C)刺激以后,MDA-MB-231细胞的增殖受到抑制,凋亡试剂盒检测显示Poly(I:C)刺激以后对细胞凋亡没有影响；实时定量PCR显示TNF-α、IFN-β和IFN-γ在Poly(I:C)刺激6h后大约有20倍的升高.结论 Poly(I:C)刺激MDA-MB-231细胞后,细胞增殖受到抑制,但其凋亡并不受影响；TNF-α、IFN-β和IFN-γ可能在抑制增殖的过程中发挥一定的作用.
目的 Toll樣受體(TLRs)在腫瘤的髮生髮展和腫瘤免疫中起到重要的作用,有報道稱TLR3的激活能夠促進某些腫瘤細胞的凋亡.本研究旨在探討TLR3的特異性激動劑Poly(I:C)對噁性程度較彊的乳腺癌細胞MDA-MB-231增殖與凋亡的影響,併初步探討其髮揮作用的機製.方法 反轉錄PCR檢測Toll樣受體各箇亞型在mRNA水平上的錶達；CCK-8細胞活力試劑盒檢測不同濃度Poly(I:C)對MDA-MB-231細胞增殖的影響；流式細胞術用Annexin V-FITC/PI染色檢測Poly(I:C)對MDA-MB-231細胞凋亡作用的影響；實時定量反轉錄PCR檢測相關細胞因子的錶達變化.結果 在mRNA水平上,MDA-MB-231細胞TLRs各亞型中TLR3錶達最高；CCK-8檢測結果顯示Poly(I:C)刺激以後,MDA-MB-231細胞的增殖受到抑製,凋亡試劑盒檢測顯示Poly(I:C)刺激以後對細胞凋亡沒有影響；實時定量PCR顯示TNF-α、IFN-β和IFN-γ在Poly(I:C)刺激6h後大約有20倍的升高.結論 Poly(I:C)刺激MDA-MB-231細胞後,細胞增殖受到抑製,但其凋亡併不受影響；TNF-α、IFN-β和IFN-γ可能在抑製增殖的過程中髮揮一定的作用.
목적 Toll양수체(TLRs)재종류적발생발전화종류면역중기도중요적작용,유보도칭TLR3적격활능구촉진모사종류세포적조망.본연구지재탐토TLR3적특이성격동제Poly(I:C)대악성정도교강적유선암세포MDA-MB-231증식여조망적영향,병초보탐토기발휘작용적궤제.방법 반전록PCR검측Toll양수체각개아형재mRNA수평상적표체；CCK-8세포활력시제합검측불동농도Poly(I:C)대MDA-MB-231세포증식적영향；류식세포술용Annexin V-FITC/PI염색검측Poly(I:C)대MDA-MB-231세포조망작용적영향；실시정량반전록PCR검측상관세포인자적표체변화.결과 재mRNA수평상,MDA-MB-231세포TLRs각아형중TLR3표체최고；CCK-8검측결과현시Poly(I:C)자격이후,MDA-MB-231세포적증식수도억제,조망시제합검측현시Poly(I:C)자격이후대세포조망몰유영향；실시정량PCR현시TNF-α、IFN-β화IFN-γ재Poly(I:C)자격6h후대약유20배적승고.결론 Poly(I:C)자격MDA-MB-231세포후,세포증식수도억제,단기조망병불수영향；TNF-α、IFN-β화IFN-γ가능재억제증식적과정중발휘일정적작용.
Objective Toll-like receptors (TLRs) play important role in the progression and tumor immunity of some types of cancer,some research have demonstrated that agonist of TLR3 can trigger apoptosis of cancers.This study was proposed to investigate if Poly(I:C),the specific agonist of TLR3,could impact proliferation or apoptosis of progressive breast cancer cells MDA-MB-231,and to investigate the primary mechanism of the function.Methods Expression of TLR1-10 mRNA was detected by quantitative real-time reverse transcription-polymerase chain reaction.Cell Counting Kit-8 was used to determine the inhibitory effect of Poly(I:C) on proliferation of MDA-MB-231 cells.Cell apoptosis was assayed by flow cytometry with V-FITC/PI staining.Results First,the toll-like receptors 1-10 were all expressed on MDA-MB-231 cells,while the expression level of TLR8 was lower than that of others.Second,according to the CCK-8,the proliferation of MDA-MB-231 cells was inhibited,but the apoptosis was not affected on the basis of Apoptosis Kit.At last,the mRNA expression of TNF-α、IFN-β and IFN-γ were elevated approximately 20 times after Poly(I:C) stimulation for 6 hours.Conclusion MDA-MB-231 cells express all toll-like receptors on mRNA level,and TLR8 was expressed lower than others.The stimulation of TLR3 with Poly(I:C) can inhibit the proliferation of MDA-MB-231,but had no effect on apoptosis.TNF-α、IFN-β and IFN-γ maybe participate in this process.
