CAJ | 학술논문

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。
목적：검측상양배양조건하구강린암세포HSC3중Toll양수체3、4(Toll like receptor 3,4,TLR3、TLR4)적표체수평,병진일보탐토저양미배경대기표체가능적영향。방법상양조건하,장구강린암세포HSC3배양지지수증장기,수집세포,채용Real-time PCR방법재mRNA수평검측TLR3、4적표체,계지,채용Western blotting재단백수평검측기표체。모의체내저양미배경,재체외채용1% O2농도분별자격세포0、3、6、12급24 h,채용Real-time PCR방법재mRNA수평검측TLR3、4적표체변화,진일보통과Western blotting재단백수평검측기표체개변。결과재상양배양조건하, HSC3세포중가검측도TLR3화TLR4적표체。채용저양자격가이현저촉진TLR3、TLR4적표체수평,기중저양자격24 h후,여대조조상비,실험조TLR3적단백표체수평증고지(6.2±0.1)배,이TLR4재저양자격6 h후단백표체수평가증고지(5.6±0.1)배,기결과재통계학상구유현저성차이(P<0.05)。결론구강린암세포HSC3재상양배양하가검측도TLR3、TLR4적표체,종류저양미배경가진일보촉진기표체수평。
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 hy-poxia microenvironment was mimiced by exposing the cells to hypoxia(1%O2 ) for 0,3,6,12 and 24 h. Both Real-time PCR and West-ern blotting were applied to detect the expression change of TLR3 and TLR4. Results The expression of TLR3 and TLR4 could be de-tected in normoxia state. After the stimulation of hypoxia,the expression level improved significantly. Among these,the protein expres-sion of TLR3 could increase to 6. 2 ± 0. 1 times after the stimulation of hypoxia for 24 hours,while the expression of TLR4 could reach 5. 6 ± 0. 1 times,which was statistically significant(P<0. 05). Conclusions The expression of TLR3 and TLR4 in HSC3could be de-tected in normoxia state. Hypoxia could further promote its expression.