趋化因子受体6／趋化因子20生物轴在肝癌组织中的表达及临床意义

Expression and its clinical significance of chemokine receptor 6/chemokine ligand 20 axis in hepatoeeUular carcinoma

目的探讨趋化因子受体6／趋化因子20生物轴在肝癌组织中的表达情况和临床意义。方法选择2003年3月至2005年12月肝癌患者48例，取其肝癌组织、癌旁组织配对标本各1份。另选取8例肝良性病变患者，取其手术切除的正常肝组织各1份。用实时荧光定量PCR法检测趋化因子受体6和趋化因子20mRNA相对表达量。用免疫组织化学染色法检测趋化因子受体6和趋化因子20蛋白的表达情况。计量资料组间差异行单因素方差分析，率的比较行卡方检验，采用Spearman法计算相关系数，采用Kaplan-Meier法绘制生存曲线，生存率的比较行Log-rank检验。结果肝癌组织的趋化因子受体6和趋化因子20的mRNA相对表达量（0．99±0．21和0．46±0．11）均高于癌旁组织（0．33±0．09和0．31±0．07）和正常肝组织（0．22±0．06和0．28±0．05），差异均有统计学意义（F=127．43和21．10，P均〈0．05）。肝癌组织趋化因子受体6蛋白阳性表达率[54．17％（26／48）]高于癌旁组织[16．67％（8／48）]和正常肝组织（0／8），差异有统计学意义（X^2=19．55，P％0．05）。肝癌组织趋化因子20蛋白阳性表达率[50．00％（24／48）]与癌旁组织[33．33％（16／48）]和正常肝组织（2／8）比较差异均无统计学意义（P均〉O．05）。肝癌组织中趋化因子受体6蛋白阳性表达率与趋化因子20蛋白阳性表达率之间存在相关性（r=0．42，P％0．05）。趋化因子受体6蛋白阳性表达率与肝癌pTNM分期、有否脉管癌栓、有否肝内转移、有否肺转移相关（X^2=5．48、4．02、5．07、5．19，P均〈0．05）。趋化因子20蛋白阳性表达率与肝癌肿瘤最大径、pTNM分期相关（X^2=4．09、4．00，P均〈0．05）。趋化因子受体6蛋白阳性组的无瘤生存率和总体生存率均低于阴性组（X^2=4．57和6．57，P均〈0．05）。趋化因子20蛋白阳性组的无瘤生存率和总体生存率与阴性组比较差异均无统计学意义（P均〉0．05）。结论趋化因子受体6／趋化因子20生物轴与肝癌的恶性生物学行为和预后相关。

Objective To explore the expression and clinical significance of chemokine receptor 6 （CCR6）/chemokine ligand 20（CCL20） axis in hepatocellular carcinoma （HCC）. Methods From March 2003 to December 2005, 48 patients with HCC were selected, and one specimen of HCC tissue and one of corresponding adjacent tissue were taken from every patient. Another eight patients with benign liver lesions were selected, and one specimen of surgical sectioned normal liver tissue of each was taken. The relative expression quantity of CCR6 and CCL20 at mRNA level was detected by real-time fluorescence quantitative polymerase chain reaction （qRT-PCR）. And the expression of CCR6 and CCL20 at protein level was determined by immunohistochemisty methods. One-way analysis of variance （ANOVA） was performed for comparison among groups of measurement data. Chi-square test was used for rate comparison. The correlation coefficient was calculated by Spearman＇ s method. Survival curves wereplotted by Kaplan-Meier method and the survival rate was compared by Log-rank test. Results The relative expression quantity of CCR6/CCL20 at mRNA level in HCC tissues （0. 99 ± 0. 21 and 0.46 ± 0.11） were significantly higher than those of para-carcinoma tissues （0.33 ±0.09 and 0.31 ± 0.07） and normal liver tissues （0.22±0.06 and 0. 28±0. 05） , and the differences were statistically significant （F= 127.43 and 21.10, both P〈0.05）. The positive percentage of CCR6 protein expression in HCC tissues （54.17%, 26/48） was significantly higher than that in para-carcinoma tissues （16. 67%, 8/48） and normal liver tissues （0/8）, and the difference was statistically significant （X^2 =19.55, P〈0.05）. There was no statistically significant difference in the positive percentage of CCL20 protein expression among HCC tissues （50.00~/0 , 24/48）, para-carcinoma tissues （33.33~, 16/48）, normal liver tissues （2/8） （all P〈0.05）. There was correlation between the positive percentage of CCR6 protein expression and that of CCL20 protein expression in HCC tissues （r= 0.42, P〈0.05）. The positive percentage of CCR6 protein expression was correlated with the pTNM stage of HCC, vascular tumor thrombosis, intrahepatic metastasis and lung metastasis （X^2 =5.48, 4.02, 5.07 and 5.19,all P〈0.05）. The positive percentage of CCL20 expression was significantly correlated to tumor maximum diameter and pTNM stage （X^2 =4.09 and 4.00,both P〈0.05）. Both the disease-free survival （DFS） rate and overall survival （OS） rate of CCR6-positive group were significantly lower than those of negative group （X^2 =4.57 and 6.57, both P〈 0.05）. There were no significant differences in DFS rate and OS rate between CCL20-positive group and negative group （both P〉0.05）. Conclusion CCR6/CCL20 axis may be related with the malignant behavior and the prognosis of HCC.