Toll样受体9和环氧合酶2在口腔鳞状细胞癌及癌旁组织中的表达意义

Expressions of TLR9 and COX-2 in oral squamous cell carcinoma and paracancerous tissues and their significance

目的探讨Toll样受体9(TLR9)和环氧合酶2(COX-2)在口腔鳞状细胞癌(OSCC)及其癌旁组织中的表达和意义。方法选择OSCC患者36例,切取OSCC组织和肿瘤边界外约5mm的癌旁组织,采用免疫组织化学法检测其中TLR9和COX-2蛋白的表达,与18例正常口腔黏膜组织对比分析。采用图像分析Image J软件对3种组织照片进行分析,计算平均吸光度(A)值;采用双评分半定量法分析3种组织切片染色强度。多组平均A值比较采用单因素方差分析,组内两两比较采用LSD-t检验;3种组织间免疫组化染色阳性率的比较采用卡方检验或Fisher精确概率法,TLR9与COX-2在OSCC组织及癌旁组织中表达的相关性分析采用Spearman法。结果OSCC组织的TLR9阳性细胞的平均A值为0.19±0.06,癌旁组织为0.18±0.05,正常组织为0.13±0.04,3种组织切片间差异具有统计学意义(F=8.022,P=0.001),OSCC组织与癌旁组织均高于正常组织(t_(OSCC)=3.99,P_(OSCC)<0.001;t_(癌旁组织)=2.88,P_(癌旁组织)=0.005)。OSCC组织COX-2阳性细胞的平均A值为0.20±0.06,癌旁组织为0.18±0.05,正常组织为0.14±0.04,3种组织切片间差异具有统计学意义(F=7.716,P=0.01),COX-2在OSCC组织中表达高于癌旁组织(t=2.00,P=0.048),在OSCC组织及癌旁组织中表达均高于正常组织,差异具有统计学意义(t_(OSCC)=3.90,P_(OSCC)<0.001;t_(癌旁组织)=2.25,P_(癌旁组织)=0.027)。TLR9与COX-2在3种组织中表达阳性率差异有统计学意义(χ^(2)_(TLR9)=45.280,P_(TLR9)<0.001;χ^(2)_(COX-2)=30.972,P_(COX-2)<0.001),癌组织和癌旁组织的TLR9阳性率均高于正常黏膜组织,差异具有统计学意义(χ^(2)_(OSCC)=30.375,P_(OSCC)<0.001;χ^(2)_(癌旁组织)=30.375,P_(癌旁组织)<0.001);癌组织和癌旁组织的COX-2阳性率高于正常黏膜组织,差异具有统计学意义(χ^(2)_(OSCC)=23.336,P_(OSCC)<0.001;χ^(2)_(癌旁组织)=17.763,P_(癌旁组织)<0.001)。TLR9和COX-2在OSCC组织及癌旁组织中的表达有相关性(r_(OSCC)=0.804,P_(OSCC)<0.001;r_(癌旁组织)=0.533,P_(癌旁组织)=0.001)。结论TLR9和COX-2在OSCC及癌旁组织中表达升高,TLR9可能通过调控其下游信号分子COX-2在OSCC的发生、发展中发挥重要作用。

Objective To investigate the expression and significance of Toll-like receptor 9(TLR9)and cyclooxygenase 2(COX-2)in oral squamous cell carcinoma(OSCC)and paracancerous tissues.Methods A total of thirty-six patients with OSCC were selected,in which OSCC tissues and paracancerous tissues about 5 mm beyond the tumor boundary were obtained.The expression of TLR9 and COX-2 protein were detected by immunohistochemistry,and compared with 18 normal oral mucosa tissues.Image analysis was done to analyze the photos of the three kinds of tissues to calculate the average absorbance(A)value.The double-scoring semi-quantitative method was used to analyze the staining intensity of the specimens.The absorbance values of multiple groups were compared by One-Way analysis of variance,and the pairwise comparisons within the groups were done by LSD-t test;the positive rate of immunohistochemical staining among the three kinds of tissues was compared by chi-square test or Fisher’s exact probability method.The Spearman method was used to analyze the correlation of TLR9 and COX-2 expression in OSCC tissues and paracancerous tissues.Results The average absorbance(A)values of TLR9 positive cells in OSCC,the paracancerous and normal tissues were 0.19±0.06,0.18±0.05 and0.13±0.04,respectively,which was significant different(F=8.022,P=0.001).Specifically,the average absorbance values of OSCC and paracancerous tissues were higher than that of normal oral mucosa tissues(t_(OSCC)=3.99,P_(OSCC)<0.001,t_(paracancerous tissues)=2.88,P_(paracancerous tissues)=0.005).The average A values of COX-2 positive cells in OSCC,the paracancerous and normal tissues were 0.20±0.06,0.18±0.05 and 0.14±0.04,respectively,which wassignificantly different(F=7.716,P=0.01).The expression of COX-2 in OSCC tissue was higher than that in the paracancerous tissues(t=2.00,P=0.048),and the expression of COX-2 in both OSC.C and paracancerous tissues was higher than that in the normal oral mucosa tissues,which was statistically different(t_(OSCC)=3.90,P_(OSCC)<0.001;t_(paracancerous tissue)=2.25,P_(paraancerous tissue)=0.027).The positive rates of TLR9 and COX-2 in the three kinds of tissues were significantly different(χ^(2)_(TLR9)=45.280,P_(TLR9)<0.001;χ^(2)_(COX-2)=30.972,P_(COX-2)<0.001).The positive rate of TLR9 in the cancerous and paracancerous tissues was higher than that of the normal oral mucosa tissues,and the difference is statistically significant(χ^(2)_(COCC)=30.375,P_(OSCC)<0.001;χ^(2)_(paracaeroue tissues)=30.375,P_(paracanceus tissues)<0.001).The positive rate of COX-2 in the cancerous and paracancerous tissues was higher than that in the normal oral mucosa tissues,which was statistically different(χ^(2)_(OSCC)=23.336,P_(OSCC)<0.001;χ^(2)_(paracanceous tissues)=17.763,P_(paracancerous tissres)<0.001).The expressions of TLR9 and COX-2 in OSCC and paracancerous tissues were correlated(r_(OSCC)=0.804,P_(OSCC)<0.001;r_(paracancerous tissues)=0.533,P_(paracancerous tissues)=0.001).Conclusions TLR9 and COX-2 were highly expressed in OSCC and paracancerous tissues.TLR9 may play an important role in the occurrence and development of OSCC by regulating its downstream signal molecule COX-2.