| 杨 潇,王艳清,程艳香.TLR4促进HIF-1α的高活性在宫颈癌中的作用机制[J].肿瘤学杂志,2016,22(12):981-987. |
| TLR4促进HIF-1α的高活性在宫颈癌中的作用机制 |
| Mechanism of TLR4 in Promoting High Activity of HIF-1α in Cervical Cancer |
| 投稿时间：2016-06-10 |
| DOI：10.11735/j.issn.1671-170X.2016.12.B001 |
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| 中文关键词: Toll 样受体 4 宫颈肿瘤 核因子?鄄κB 低氧诱导因子1α 活性氧 |
| 英文关键词:toll-like receptor 4 cervical neoplasms NF-κB hypoxia inducible factor-1α reactive oxygen species |
| 基金项目:国家自然科学基金（81302273）；湖北省卫生厅中医药科研项目（2012Z-Y02）；湖北省卫计委一般面上项目（WJ2015MB084）；湖北省科技厅科技项目支撑计划 （2015BCA313） |
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| 中文摘要: |
| 摘 要：［目的］ 研究Toll 样受体 4( toll-like receptor 4，TLR4)促进低氧诱导因子1α（hypoxia inducible factor-1α，HIF-1α）的高活性在宫颈癌中的作用及机制。［方法］ 体外培养Siha细胞，分为空白对照组（control组）、脂多糖组（LPS组）、NF-κB信号通路干预组［吡咯烷二硫氨基甲酸（PDTC）+LPS组］和TLR4信号干预组（siTLR4+LPS组）共4组，用 MTT法、软琼脂形成实验观察各组细胞的活性增殖、克隆情况；用免疫细胞化学染色法和Western blot法检测细胞内HIF-1α含量变化；DCFH-DA 法检测活性氧（ROS）含量变化；光泽精化学发光法检测NADPH氧化酶含量变化。 ［结果］ MTT和软琼脂形成实验结果显示：与control组相比，LPS组细胞活性和增殖能力增强，PDTC+LPS组无明显变化，siTLR4+LPS组降低（P<0.05）；与LPS组相比，PDTC+LPS组和siTLR4+LPS组细胞活性和增殖能力均降低（P<0.05）。与PDTC组+LPS组相比，siTLR4+LPS组细胞活性显著减弱（P<0.01）。免疫细胞化学染色和Western blot法结果显示：与control组相比，LPS组HIF-1α活性增强（P<0.05），siTLR4+LPS组活性降低（P<0.05）；与LPS组相比，PDTC+LPS组HIF-1α表达降低（P<0.05），siTLR4+LPS组HIF-1α表达明显降低（P<0.01）；与PDTC+LPS组相比，siTLR4+LPS组细胞HIF-1α活性减弱（P<0.05）。NADPH氧化酶和ROS活性检测结果显示：与control组相比，LPS组NADPH氧化酶和ROS活性增强（P<0.05），siTLR4+LPS组降低（P<0.05），LPS组与PDTC+LPS组无明显差异。与PDTC+LPS组相比，siTLR4+LPS组NADPH氧化酶和ROS活性均降低（P<0.05）。［结论］ TLR4可通过核因子-κB(NF-κB)信号途径促进宫颈癌的发生和发展，NADPH氧化酶参与TLR4维持细胞内HIF-1α高活性，而此过程与NF-κB信号无关，这为研究宫颈癌的发生发展机制提供新的思路。 |
| 英文摘要: |
| Abstract：［Objective］ To study the role and mechanism of toll-like receptor 4(TLR4) in promoting the activity of hypoxia inducible factor-1α(HIF-1α) in cervical cancer. ［Methods］ SiHa cells were cultured in vitro and divided into four groups：control group，lipopolysaccharide(LPS) group，pyrrolidine dithiocarbamate(PDTC)+LPS group and siTLR4+LPS group. Active proliferation and cloning of the cells in each group were observed by MTT method and soft agar forming test. The change of HIF-1α was detected with immunocytochemical staining and Western blot method. The change of reactive oxygen species(ROS) content and NADPH oxidase content were detected by DCFH-DA method and chemiluminescence method respectively. ［Results］ MTT and soft agar forming test showed that compared with control group，the cell activity and proliferation ability were enhanced in LPS group，PDTC+LPS group had no significant change，whereas decreased in siTLR4+LPS group(P<0.05)；compared with LPS group，the proliferation activity and cell activity in PDTC+LPS group and siTLR4+LPS group was decreased(P<0.05)；compared with the PDTC+LPS group，the cell activity of siTLR4+LPS group was significantly decreased(P<0.01).Immunocytochemical staining and Western blot showed that compared with the control group，HIF-1α activity in LPS group was enhanced(P<0.05)；HIF-1α activity in siTLR4+LPS group was decreased(P<0.05)；compared with the LPS group，HIF-1α activity in PDTC+LPS group and in siTLR4+LPS group was decreased(P<0.05 and P<0.01)；compared with the PDTC+LPS group，HIF-1α activity in siTLR4+LPS group was weakened(P<0.05). NADPH oxidase and ROS detection results showed that compared with the control group，the activity of NADPH oxidase and ROS enhanced in LPS group(P<0.05)，decreased in siTLR4+LPS group(P<0.05)；and there were no significant differences between LPS group and PDTC+LPS group；compared with the PDTC+LPS group，the NADPH oxidase and ROS activity were decreased in siTLR4+LPS group(P<0.05). ［Conclusion］ TLR4 may promote the occurrence and development of cervical cancer through NF-κB signaling pathway.NADPH oxidase may participate in the process of TLR4 to maintain the high intracelluar activity of HIF-1α and this process may be not related to NF-κB signal. |
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