阻断趋化因子受体3途径对结肠癌患者术后局部复发和肝转移影响的研究

Effect of blocking the CXC chemokine receptor 3 pathway on postoperative colonic cancer local recurrence and hepatic metastasis in a nude mouse model

目的探讨趋化因子受体3（CXCR-3）对结肠癌术后局部复发和肝转移的影响。方法常规培养HT-29细胞，采用RT-PCR、Western-blot方法检测靶向CXCR-3的反义肽核酸（asPNA）对CXCR-3 mRNA和蛋白表达水平的变化。建立人结肠癌术后局部复发裸鼠动物模型、人结肠癌术后肝转移动物模型。分别经尾静脉注射asPNA、随机错配肽核酸和生理盐水，分析CXCR-3的asPNA对结肠癌术后局部复发和肝转移的抑制作用。结果asPNA处理结肠癌细胞后，CXCR-3 mRNA和蛋白表达受到明显抑制（P〈0．05）。asPNA组与随机错配组和对照组比较，裸鼠结肠癌术后复发率（20％比60％、60％，P〈0．05）、肝转移率（30％比100％、100％，P〈0．05）、平均肝转移瘤数目[（7．6±2．4）比（28．4±9．8）、（26．8±8．6），P〈0．05]均显著降低。结论CXCR-的asPNA能够抑制结肠癌术后局部复发和肝转移，其机制可能与其特异性抑制HT-29细胞CXCR-3 mRNA和蛋白表达有关。

Objective To identify the effect of blocking the CXC chemokine receptor 3 pathway on postoperative local recurrence and live metastasis of colon cancer in a nude mouse model. Methods Cultured HT-29 cells were devided into three groups： control group, antisense peptide nucleic acid group（asPNA） and mismatch PNA group. After 24 h of transfection, cultured cells were harvested to detect the changes of CXCR-3 mRNA and protein by RT-PCR and Western-blot respectively. Local recurrence model and liver metastasis model of human colon cancer in nude mouse were constructed Saline, asPNA and mismatch PNA was injected via tail vein respectively. The local recurrence and liver metastasis rate, the mean live metastasis number were analysed. Results Compared with other two groups asPNA down regulated the expression of CXCR-3 mRNA（0. 56±0. 10） vs. （ 1.38±0. 13） vs. （ 1.06±0. 12） ,P 〈 0. 05 and protein （ 2. 88 ± 1.60） vs. （6.48 ± 2. 56） vs. （ 5.86 ± 2. 30） , P 〈 0. 05. Tumor local recurrence rate （20% vs. 60% vs. 60%, P〈0.05）, liver metastasis rate（30% vs. 100% vs. 100%, P〈0.05） , and mean live metastasis number [（7.6±2.4） vs. （28.4±9.8） vs. （26.8±8.6）, P〈0.05] were significantly decreased by asPNA. Conclusion By decreasing expression of CXCR-3 mRNA and protein, asPNA targeting at CXCR-3 inhibits postoperative local recurrence and live metastasis of colon cancer.