TLR4和NF-κB p65表达与结肠癌的关系

The expression and correlation of TLR4 and NF-κB p65 in colorectal carcinoma

目的 评价结肠癌和癌旁正常组织中TLR4 mRNA和NF-κB p65蛋白表达水平与结肠癌发生发展的相关性.方法 取手术切除的新鲜结肠癌组织和距癌组织2 cm以上的癌旁组织各63份,其中：结肠癌高分化型组23份,中分化型组17份,低分化型组20份,其他分化类型3份;伴淋巴结转移组27份,不伴淋巴结转移组36份;Dukes A期组18份,Dukes B期组14份,DukesC期组22份,Dukes D期组9份.采用实时荧光定量RT-PCR检测结肠癌组织和癌旁组织中TLR4 mRNA的表达水平;用WB法检测NF-κB p65蛋白的相对表达水平.结果 结肠癌组织中TLR4 mRNA表达量为86.42±15.16,明显高于癌旁组织的32.74±9.44,差异有统计学意义（t=22.354,P＜0.01）.高、中、低分化型结肠癌TLR4 mRNA表达量分别为69.58±11.27、64.57±13.91、97.12±15.44,低分化型结肠癌表达量高于高、中分化型结肠癌（t值分别为11.304、12.223,P均＜0.01）.伴淋巴结转移组TLR4 mRNA表达量（89.91±13.33）与不伴淋巴结转移组（81.16±13.59）比较,差异无统计学意义（t=0.959,P＞0.05）.Dukes A期组TLR4 mRNA表达量（59.05±11.66）低于Dukes B～D期组的表达量（分别为92.32±17.51、91.41±15.21、101.46±17.43）,差异有统计学意义（t值分别为8.708、9.664、9.525,P均＜0.05）;结肠癌组织和癌旁组织NF-κB p65蛋白表达量分别为0.63±0.11、0.34±0.08,结肠癌组织NF-κB p65表达量高于癌旁组织（t=18.266,P＜0.01）,高、中、低分化型结肠癌组织中NF-κB p65蛋白表达量分别为0.46±0.09、0.72±0.11、0.77±0.14,中低分化型组表达高于高分化型（t值分别为11.223、10.875,P均＜0.01）,伴淋巴结转移组与不伴淋巴结转移组NF-κB p65表达量分别为0.82±0.17、0.57±0.12,且差异有统计学意义（t=18.269,P＜0.05）.Dukes A期的NF-κB p65表达量（0.39±0.06）低于Dukes B～D期（分别为0.72±0.12、0.69±0.14、0.76±0.13）,且差异有统计学意义（t值分别为10.442、9.889、9.721,P均＜0.01）.结论 TLR4/NF-κB p65信号通路的表达水平升高与结肠癌的临床进展和病理分级密切相关;NF-κB p65可能是结肠癌转移的分子指标之一,TLR4/NF-κB p65升高可促进结肠癌的发生发展.

Objective To investigate the expression of TLR4 mRNA and NF-κB p65 in colorectal carcinomas and adjacent normal colon tissue, and evaluate their roles in the pathogenesis and development in colorectal carcinoma. Methods Sixty-three colorectal carcinoma samples and respective adjacent normal colon tissue samples （ well differentiated ： 23 cases; moderately differentiated： 17 cases; poorly differentiated：20 cases; other differentiated type： 3 cases; lymph node metastasis： 27 cases; no lymph node metastasis：36 cases; Dukes A： 18cases;Dukes B： 14 cases Dukes C： 22 cases; Dukes D： 9 cases） were collected. The expression of TLR4 mRNA in colorectal carcinomas and adjacent tissue were detected by RT-PCR. The expression of NF-κB p65 was detected by WB. Results The expression of TLR4 mRNA in colorectal carcinomas and adjacent tissue were 86.42 ± 15.16 and 32.74 ± 9.44. It was significantly higher in carcinoma tissue than that in adjacent tissue （ t = 22.354, P 〈 0.01 ）. The expression of TLR4 mRNA in well, moderately and poorly differentiated coiorectal carcinomas were 69.58 ± 11.27, 64.57 ± 13.91 and 97.12 ± 15.44 respectively. TLR4 mRNA in poorly differentiated colorectal carcinomas was significantly higher than that in well, moderately differentiated ones （ t = 11.304 and 12.223, P 〈 0.01 ）. There was no difference between lymph node metastatic carcinomas （ 89.91 ± 13.33 ） and carcinomas without metastasis （81.16±13.59,t =0.959,P〉0.05）. The expression of TLR4 mRNA in the Dukes A stage tumors （59.05±11.66） was lower than that in Dukes B（90.34 ±0.08）,C（91.41 ± 15.21）, D（101.46 ±17.43）, respectively （ t = 8.708,9.664,9.525, P 〈 0.05 ）. The expression of NF-κB p65 in colorectal carcinoma（0.63 ±0.11） was significant higher than that in adjacent tissue（0.34 ±0.08,t = 18.266,P 〈0.01 ）. The expression of NF-κB p65 in well, moderately and poorly differentiated colorectal carcinomas were 0.46 ± 0.09, 0.72 ± 0.11 and 0.77 ± 0.14, respectively. The experssion of NF-κB p65 in well differentiated colorectal carcinomas was obviously lower than the woderately and poorly differentiated carcinomas （t = 11.223 and 10.875, P 〈0.01 ）. There was significant difference between the expression of p65 in lymph node metastatic carcinomas（0.82 ± 0.17） and non-metastatic carcinomas（0.57 ± 0.12, t =18.269,P〈0.05）. The expression of NF-κB p65 in Dukes A colorectal carcinomas （0.39 ± 0.06） was lower compared with the Dukes B（0.72 ±0.12）, C（0.69 ±0.14） and D carcinomas（0.76 ±0.13,t =10.442, 9.889 and 9.721, P 〈 0.01 ）. Conclusions The enhanced expression of TLR4/NF-κB p65 are closely associate with clinical stage and pathologic grade. NF-κB p65 may be an molecular marker of lymph node metastatic. The increased expression of TLR4/NF-κB p65 promote the pathogenesis and development of colorectal carcinoma.