Toll样受体4活化增强乳腺癌细胞株MCF-7侵袭力的研究
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江苏省检验医学重点实验室开放课题资助(JSKLM-2014-004)


Stimulation of toll-like receptor 4 promotes human breast cancer cell MCF-7 invasiveness
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    摘要:

    目的:研究Toll样受体4(TLR4)在人类乳腺癌MCF-7细胞株中的表达及在肿瘤增殖和转移中的作用-方法:脂多糖(LPS) 刺激MCF-7细胞株,RT-PCR-Real-time PCR-FCS 和 Western blotting 检测TLR4在mRNA和蛋白水平表达变化;MTT检测细胞增殖;RT-PCR 和 real-time PCR检测基质金属蛋白酶(MMP-2)-MMP-9 和血管内皮生长因子(VEGF)的mRNA表达并用ELISA检测培养上清中其蛋白表达水平;Western blot检测TLR4信号传导通路下游蛋白MyD88的表达;CBA测定细胞培养上清的炎性细胞因子;划痕实验检测癌细胞的生物学侵袭力-结果:实验表明,LPS刺激MCF-7细胞株能明显上调TLR4 mRNA和蛋白表达(P < 0.05)-MTT结果显示,LPS对癌细胞增殖没有影响-LPS激活TLR4后,MMP-2-MMP-9和VEGF在mRNA和蛋白水平的表达均明显上调 (P < 0.05)-划痕实验表明,LPS能显著增强MCF-7细胞株的划痕愈合能力-此外,LPS能诱导TLR4信号通路下游MyD88蛋白表达的显著上调(P < 0.05),IL-6的分泌增多(P < 0.05)-结论:LPS能明显增强MCF-7细胞株TLR4的表达,增强癌细胞的侵袭力-

    Abstract:

    Objective:This study investigated the expression and biological role of toll-like repeptor-4 (TLR4) in human breast cancer proliferation and metastasis. Methods:Lipopolysaccharide (LPS) was used to stimulate MCF-7,expression of TLR4 mRNA and protein was analyzed by RT-PCR,Real-time PCR,FCS and Western blotting;MMP-2,MMP-9 and VEGF were measured by RT-PCR and Real-time PCR on the mRNA levels and ELISA on the protein levels;MyD88 was analyzed by Western blotting;The cytokines in the supernatants were measured by CBA;The cell proliferation was analyzed by MTT;The invasion was analyzed by wound healing assay. Results:Using LPS to stimulate MCF-7,TLR4 mRNA and protein increased obviously. However,the proliferation had no significant changes. In addition,the activation of TLR4 promoted the expression of MMP-2,MMP-9 and VEGF notablely. LPS promoted the invasion of MCF-7. Moreover,the stimulation of LPS significantly increased the expression of MyD88,and caused the secrection of IL-6. Conclusion:This study indicated that TLR4 may participate in the progression and metastasis of human breast cancer.

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王 涛,王 波,温会燕,颜 洁,何春燕,朱雪明,杨 欢. Toll样受体4活化增强乳腺癌细胞株MCF-7侵袭力的研究[J].南京医科大学学报(自然科学版),2016,(5):533-538

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  • 收稿日期:2016-01-13
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  • 在线发布日期: 2016-05-23
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