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目的从基因水平评估“不接触分离技术”是否对结直肠癌术中癌细胞门静脉播散起抑制作用。方法应用突变等位基因特异性扩增法(MASA),检测结直肠癌术中处理肿瘤前、中、后期的门静脉血中癌细胞播散情况。46例结直肠癌患者中,22例施行传统手术,24例应用“不接触分离技术”。在打开腹腔处理肿瘤前,经胃网膜右静脉在门静脉中置入一静脉导管,然后分别于处理肿瘤前、中、后期经导管各采血10ml。用MASA法检测肿瘤特异性标记物———K-ras的12、13、61密码子和P53的5、6、7、8外显子,并用3’末端20bp的寡核苷酸作为PCR的引物。结果22例接受传统手术的患者中,15例体细胞有突变,其中12例(80.0%)术中在门静脉血中用MASA法检测到原发肿瘤细胞特异性标记物;而在24例“不接触分离技术”中,有17例发现有K-ras和P53基因的突变,其中仅2例(11.8%)在术中于门静脉血中检测到原发肿瘤细胞特异性标记物;显示传统手术操作与癌细胞播散呈正相关(P<0.05)。经随访,传统手术组术后2年内,有5例出现肝转移,并有4例死亡;而“不接触分离技术”组,术后2年内仅1例出现肝转移并死亡。结论与传统手术相比,“不接触分离技术”能抑制结直肠癌手术操作过程中癌细胞的门静脉播散,有效地降低结直肠癌的肝转移率,提高患者的2年生存率。
Objective To evaluate whether “non-contact separation technique” plays an inhibitory role in the dissemination of portal vein of cancer cells during colorectal cancer surgery at the gene level. Methods The mutation allele-specific amplification (MASA) method was used to detect the dissemination of cancer cells in the portal vein before, during and after the operation of colorectal cancer. Of the 46 patients with colorectal cancer, 22 underwent conventional surgery and 24 underwent “non-contact separation.” Before opening the abdominal cavity to treat the tumor, a venous catheter was placed in the portal vein through the right gastric vein and then 10 ml of each catheter was collected before, during and after the treatment of the tumor. The tumor-specific marker, the 12,13,61 codon of K-ras and the 5,6,7,8 exons of P53 were detected by the MASA method and the 3’-end 20 bp oligonucleotide was used as a PCR Primer. Results Of the 22 patients who underwent conventional surgery, 15 had somatic cell mutations. Twelve of them (80.0%) detected primary tumor cell-specific markers in the portal vein using intraoperative MASA. In 24 patients, Contact Separation Technology “, 17 cases were found to have K-ras and P53 gene mutations, of which only 2 (11.8%) intraoperative detection of primary tumor cell-specific markers in the portal vein; showed that the traditional surgical procedures And cancer cell dissemination was positively correlated (P <0.05). After follow-up, 5 patients had liver metastases and 4 patients died within 2 years after operation in the traditional operation group. Only 1 patient died of liver metastasis within 2 years after operation. Conclusion Compared with traditional surgery, ”non-contact separation technique" can inhibit the dissemination of portal vein of cancer cells during colorectal cancer operation and effectively reduce the hepatic metastasis rate of colorectal cancer and improve the 2-year survival rate of patients.