CD133、CXCR4共表达与胆囊癌转移特性的相关性研究

Correlation between co-expression of CD133,CXCR4 and metastasis ability in gallbladder cancer

目的探讨CD133、CXCR4共表达与胆囊癌转移特性之间的关系。方法培养胆囊癌GBC^-SD细胞,并经免疫磁珠分选法分选得到CXCR4^+和CXCR4^-亚群胆囊癌GBC^-SD细胞,分为CD133^-CXCR4^-、CD133^-CXCR4^+、CD133^+CXCR4^-及CD133^+CXCR4^+四组,以Transwell法检测四组胆囊癌GBC^-SD细胞侵袭能力。Western^-blot法检测CD133^+CXCR4^+组与CD133^+CXCR4^-组细胞上皮间质转化相关因子表达。Western blot法检测21例胆囊癌原发灶组织标本中CD133与CXCR4蛋白表达与淋巴结转移、肝脏侵犯、胆管侵犯及TNM分期之间的关系。结果 CD133^+CXCR4^+组平均穿膜细胞数为(32.49±9.45),显著高于CD133^+CXCR4^-组的(16.17±8.55),差异具有统计学意义(P=0.02)。CD133^+CXCR4^+组N^-cadherin蛋白灰度值为(0.8321±0.1259),显著高于CD133^+CXCR4^-组的(0.4572±0.0775),差异有统计学意义(P=0.004)。CD133^+CXCR4^+组E^-cadherin蛋白灰度值为(0.4712±0.0736),显著低于CD133^+CXCR4^-组的(0.8439±0.1339),差异有统计学意义(P=0.006)。CD133^+CXCR4^+组Snail蛋白相对灰度值为(0.9455±0.1228),显著高于CD133^+CXCR4^-组的(0.3219±0.0934),差异有统计学意义(P=0.003)。胆囊癌标本淋巴结转移组、肝脏侵犯组、胆管侵犯组CXCR4蛋白表达值显著高于对应阴性组(P=0.013,P=0.024,P=0.037),Ⅰ/Ⅱ分期组显著低于Ⅲ/Ⅳ组,差异有统计学意义(P=0.016)。淋巴结转移组CD133蛋白表达显著高于对应阴性组,差异有统计学意义(P=0.009),Ⅰ/Ⅱ分期组显著低于Ⅲ/Ⅳ组,差异有统计学意义(P=0.018)。肝脏侵犯组、胆管侵犯组与相应对照组CD133蛋白相对表达差异无统计学意义(P=0.443,P=0.371)。CD133蛋白与CXCR4蛋白表达存在明显正相关(r=0.693,P<0.01)。结论 CD133、CXCR4共表达预示胆囊癌具有较强的转移特性。

Objective To explore the correlation between co^- expression of CD133,CXCR4 and the metastasis ability in gallbladder cancer. Methods Theinvasive ability in four groups of CD133^- CXCR4^-,CD133^- CXCR4~＋ ,CD133~＋ CXCR4^-,CD133~＋ CXCR4~＋ wasmesured using Transwell. EMT related factor proteins of CD133~＋ CXCR4^- group and CD133~＋ CXCR4~＋ group were detected using Western blot. The relationship between the expression of CD133 and CXCR4 proteinsin 21 cases of primary gallbladder cancer tissue sampleswith lymph node metastasis,liver metastasis or bile duct metastasiswas analyzed by Western blot. Results The number of migrating cells in CD133~＋ CXCR4~＋ group（ 32. 49 ± 9. 45） wassignificantly greater than in CD133~＋ CXCR4^- group（ 16. 17 ± 8. 55,P = 0. 02）. The expression level of N^- cadherin protein in CD133~＋ CXCR4~＋ group（ 0. 8321 ± 0. 1259） was significantly higher than that in CD133~＋ CXCR4^- group（ 0. 4572 ± 0. 0775,P = 0. 004）. The expression level of E^- cadherin protein in CD133~＋ CXCR4~＋ group（ 0. 4712 ± 0. 0736） was significantly lower than that in CD133~＋ CXCR4^- group（ 0. 8439 ± 0. 1339,P= 0. 006）. The expression level of snail protein in CD133~＋ CXCR4~＋ group（ 0. 9455 0. 1228） was significantly higher than in CD133~＋ CXCR4^- group（ 0. 3219 ± 0. 0934,P = 0. 003）. The expression level of CXCR4 protein in the groups of lymph node metastasis,liver metastasis and bile duct metastasis was significantly higher than in control group（ P = 0. 013,P = 0. 024,P = 0. 037,respectively）. CXCR4 in stage Ⅰ/Ⅱ（ TNM） group was lower than in Ⅲ/Ⅳ group（ P = 0. 016）. The expression level of CD133 protein in the group of lymph node metastasis was significantly higher than in control group（ P = 0. 009）. The expression of CD133 in stage Ⅰ/Ⅱ（ TNM） group was lower than in Ⅲ/Ⅳ group（ P = 0. 018）. There was no significant difference between groups of liver metastasis and bile duct metastasis with control group（ P = 0. 443,P = 0.371）. The expression level of CD133 protein was positively correlated with CXCR4 protein（ r = 0. 693） withthe difference being statistically significant（ P〈0. 01）. Conclusions Co^- expression of CD133 and CXCR4 may indicate stronger capability for metastasis in gallbladder cancer.