低氧促进口腔鳞癌细胞TLR3及TLR4表达的相关研究  

Study on the expression of TLR3 and TLR4 with the stimulation of hypoxia in OSCC cell lines

作　　者：江琳琳[1] 韩伟[2] 王志勇[2] 张全安[1] 郑勤[1] 

机构地区：[1]东南大学附属第二医院肿瘤科,江苏南京210003 [2]南京市口腔医院口腔颌面外科,江苏南京210008

出　　处：《口腔医学》2015年第10期806-809,共4页Stomatology

基　　金：国家自然基金(81302351);江苏省自然基金(BK2012075;BK20131080);南京市杰出青年基金项目(JQX14010)

摘　　要：目的检测常氧培养条件下口腔鳞癌细胞HSC3中Toll样受体3、4(Toll like receptor 3,4,TLR3、TLR4)的表达水平,并进一步探讨低氧微环境对其表达可能的影响。方法常氧条件下,将口腔鳞癌细胞HSC3培养至指数增长期,收集细胞,采用Real-time PCR方法在mRNA水平检测TLR3、4的表达,继之,采用Western blotting在蛋白水平检测其表达。模拟体内低氧微环境,在体外采用1%O2浓度分别刺激细胞0、3、6、12及24 h,采用Real-time PCR方法在mRNA水平检测TLR3、4的表达变化,进一步通过Western blotting在蛋白水平检测其表达改变。结果在常氧培养条件下,HSC3细胞中可检测到TLR3和TLR4的表达。采用低氧刺激可以显著促进TLR3、TLR4的表达水平,其中低氧刺激24 h后,与对照组相比,实验组TLR3的蛋白表达水平增高至(6.2±0.1)倍,而TLR4在低氧刺激6 h后蛋白表达水平可增高至(5.6±0.1)倍,其结果在统计学上具有显著性差异(P<0.05)。结论口腔鳞癌细胞HSC3在常氧培养下可检测到TLR3、TLR4的表达,肿瘤低氧微环境可进一步促进其表达水平。Objective To investigate the expression of Toll like receptor 3,4（ TLR3,4） in oral squamous cell carcinoma（ OSCC）cells lines cultured in normoxia,and to further investigate the effect of tumor hypoxia microenvironment on its expression. Methods OSCC cell line HSC3 was cultured under normoxia conditions. Afterreachingthe exponential growth phase,the cells were collected and Real-time PCR and Western blotting were applied to detect the expression of TLRs at mRNA and protein levels respectively. Tumor hypoxia microenvironment was mimiced by exposing the cells to hypoxia（ 1% O2） for 0,3,6,12 and 24 h. Both Real-time PCR and Western blotting were applied to detect the expression change of TLR3 and TLR4. Results The expression of TLR3 and TLR4 could be detected in normoxia state. After the stimulation of hypoxia,the expression level improved significantly. Among these,the protein expression of TLR3 could increase to 6. 2 ± 0. 1 times after the stimulation of hypoxia for 24 hours,while the expression of TLR4 could reach5. 6 ± 0. 1 times,which was statistically significant（ P〈0. 05）. Conclusions The expression of TLR3 and TLR4 in HSC3 could be detected in normoxia state. Hypoxia could further promote its expression.

关 键 词：口腔鳞状细胞癌 Toll样受体3、4 低氧 

分 类 号：R739.85[医药卫生—肿瘤]