论文部分内容阅读
目的探讨分支DNA(b-DNA)及半定量RT-PCR(SqRT-PCR)在结直肠癌术中腹腔冲洗液中游离癌细胞检测中的应用。方法分别采用基于b-DNA信号放大的基因表达定量检测技术及SqRT-PCR方法检测48例结直肠癌患者术中腹腔冲洗液中CEA mRNA的表达,同时行腹腔冲洗液细胞学检查(peritoneal lavage cytolo-gy,PLC),收集12例结直肠良性病变患者的腹腔冲洗液为阴性对照,GAPDH mRNA为内参对照。结果b-DNA技术和SqRT-PCR方法检测游离癌细胞的阳性率(43.8%,31.3%)较PLC的检出率(4.2%)高(P<0.01)。结直肠癌患者腹腔冲洗液中CEA mRNA的相对表达量均与肿瘤分化程度、浆膜侵犯程度和Dukes分期有关(P<0.05),而与肿瘤大小、患者性别、年龄无关(P>0.05)。结论b-DNA技术和SqRT-PCR方法检测游离癌细胞各有优缺点;腹腔内游离癌细胞的存在与结直肠癌临床病理因素有关。
Objective To investigate the application of branched-chain DNA (b-DNA) and semi-quantitative RT-PCR (SqRT-PCR) in the detection of free cancer cells in peritoneal washings during colorectal cancer surgery. Methods The expression of CEA mRNA in peritoneal wash fluid of 48 patients with colorectal cancer was detected by quantitative gene expression detection method and SqRT-PCR based on b-DNA signal amplification. Meanwhile, the peritoneal lavage cytolo -gy, PLC), 12 cases of colorectal benign lesions were collected in patients with peritoneal washings negative control, GAPDH mRNA as an internal control. Results The positive rate (43.8%, 31.3%) of free cancer cells detected by b-DNA and SqRT-PCR was higher than that of PLC (4.2%) (P <0.01). The relative expression of CEA mRNA in peritoneal washings of colorectal cancer patients was correlated with tumor differentiation degree, serosal invasion degree and Dukes stage (P <0.05), but not with tumor size, sex and age (P> 0.05). Conclusion Both b-DNA and SqRT-PCR methods can detect the advantages and disadvantages of free cancer cells. The existence of intraperitoneal free cancer cells is related to the clinicopathological factors of colorectal cancer.