SYBR GreenⅠ实时荧光定量PCR检测结直肠癌中Tiam-1和HPA-1基因的表达

Expression of Tiam-1 mRNA and HPA-1 mRNA in the Colorectal Cancer by SYBR Green Ⅰ Real-Time Quantitative PCR

目的:检测Tiam-1基因和HPA-1基因在结直肠癌组织中的表达水平,探讨两者与结直肠癌侵袭转移及预后的关系。方法:选取2009年8月-2010年1月我院行手术治疗的结直肠癌患者53例。采用SYBR GreenⅠ实时荧光定量逆转录聚合酶链反应(RT-PCR)方法检测Tiam-1 mRNA及HPA-1 mRNA在结直肠癌组织、癌旁组织(距癌2cm)及远离结直肠癌的手术切缘正常组织中(距癌5cm以上)的表达水平,并分析两者与结直肠癌临床病理参数的关系。结果:HPA-1 mRNA和Tiam-1 mRNA在结直肠癌中的表达水平(35.87±12.30,40.79±9.32)显著高于癌旁组织(13.13±6.71,12.39±5.38)和正常组织(4.06±3.55,6.09±4.86,P均<0.05),癌旁组织中二者的表达水平高于正常组织,差异有统计学意义(P<0.05)。Tiam-1和HPA-1的基因表达水平在肿瘤的高/中分化和低分化之间,T1～T2和T3～T4之间,有淋巴结转移和无淋巴结转移之间、有血行转移和无血行转移之间及Ⅰ～Ⅱ和Ⅲ～Ⅳ期之间的差异均有统计学意义(P均<0.05)。Spearman等级相关分析证明了Tiam-1与HPA-1的表达呈正相关趋势(OR=0.46,P=0.048)。结论:Tiam-1 mRNA和HPA-1 mRNA在结肠癌组织中呈高表达,且二者的表达增强可促进结肠癌的侵袭转移,联合检测Syndecan-1及HPA-1对于指导结肠癌临床治疗及判断预后有重要价值。

Objective： To investigate the expression of Tiam-1 mRNA and HPA-1 mRNA in colorectal neoplasm,and to investigate their relativity with invasion and metastasis of colorectal neoplasm. Methods： From Aug. 2009 to Jan. 2010,53 consecutive patients with colorectal neoplasm admitted to our hospital for surgical treatment were included into this study. SYBR GreenⅠ Real-time fluorescent quantitative reverse transcriptase polymerase chain reaction（RT-PCR） was used to investigate the expression of Tiam-1 mRNA and HPA-1 mRNA in colorectal neoplasm,paired adjacent colorectal neoplasm（2 cm from carcinoma） ,surgical marginal normal colorectal mucosa tissues（5 cm from carcinoma） ,and to analyze their correlation with clinic pathological characteristics of colorectal neoplasm. Results： The expression of HPA-1 mRNA and Tiam-1 mRNA were significantly higher in colorectal neoplasm（35.87 ±12.30,40.79 ±9.32） than that in paired adjacent colorectal neoplasm（13.13±6.71,12.39±5.38） and normal colorectal mucosa（4.06±3.55,6.09±4.86,P0.05） . And the expression of the two were higher in paired adjacent colorectal neoplasm than that in normal colorectal mucosa tissues（P0.05） . Accompanied with differentiated degree of histological type descending,depth of invasion increasing,lymph node metastasis,vessel invasion and stage of TNM upgrading happening,the expression of Tiam-1 and HPA-1 significantly different. The Spearman rank correlation analysis showed a trend toward correlation between Tiam-1 and HPA-1（OR=0.46,P=0.048） . Conclusion： The expression of HPA-1 mRNA and Tiam-1 were high in colorectal neoplasm,and the expression of the two can predict the invasion and metastasis of colorectal neoplasm. Determination of Tiam-1 and HPA-1may be of value in the treatment as well as in the prognostication of colorectal neoplasm.