【作者】 王贵英； 于跃明； 马志强； 何景利； 于滨； 张振亚； 安广权；

【Author】 Wang Gui-Ving Yu Yue-Ming Ma Zhi-Qiang et al. Department of Surgery, The Fourth Hospital of Hebei medical university & Hebei provincial tumor hospital, Shijiazhuang, 050011, China

【机构】 河北医科大学第四医院暨河北省肿瘤医院外科；

【摘要】 目的:端粒酶高表达在大肠癌的发生发展中发挥着重要作用,但其在大肠癌中的表达特性还存在很多争议。为了探讨端粒酶活性与大肠癌发生及各临床病理参数的关系, 本研究应用端粒重复序列扩增和微孔板反向杂交法对40例大肠癌组织的端粒酶活性进行了研究。方法:选取2004年11 月～2005年3月在河北医科大学第四医院外二科手术治疗的大肠癌患者40例,其中男性17例,女性23例,年龄29～76 岁,平均年龄54岁。结肠癌11例,直肠癌29例。细胞分化程度:高分化腺癌23例,中分化腺癌10例,低分化腺癌7例。 Dukes分期:A期9例,B期12例,C期12例,D期7例。无淋巴结转移23例,有淋巴结转移17例。另选10例大肠良性病变患者作为对照。提取端粒酶,进行端粒重复序列扩增,端粒酶活性检测试剂盒购自华美生物工程公司,以试剂盒提供的阳性对照物作为阳性对照,以裂解液作为阴性对照,以双蒸水作为空白对照。用微孔板反向杂交法进行产物杂交,用 U-2000型分光光度计测定杂交产物在波长450nm时的吸光度(OD值),以空白对照杂交产物校零。阴性对照杂交产物OD 值低于0．05时按0．05计算,高于0．05时按实际OD值计算。样本杂交产物OD值大于或等于阴性对照杂交产物OD值的 2．1倍时,判为端粒酶活性阳性。结果:40例大肠癌组织标本中,检出端粒酶活性34例,阳性率85．0%;10例正常大肠粘膜组织标本中,均未检出端粒酶活性。两组阳性率比较差异有显著性(P<0．05)。端粒酶活性与大肠癌生长部位、癌细胞分化程度、Dukes临床肿瘤分期以及有无淋巴结转移无关。结论:端粒酶活性的激活发生在大肠癌早期,端粒酶的活性检测对结直肠癌患者的预后判断作用甚微。更多还原

【Abstract】 Objective: High expression of telomerase plays important roles in the occurrence and development of colorectal carcinoma. However, the properties of telomerase expression in colorectal cancer still have many convercial opinions. In order to investigate the relationship between telomerase activity and the pathoclinical parameters, telomeric repeat amplification protocol assay (TRAP) was imployed to detect the telomerase activity in tissues of 40 cases of colorectal carcinoma. Methods: Fourty patients with colorectal cancer who accept operation in the second Department of Surgery, Fourth Hospital of HeBei Medical University were selected from November 2004 to March 2005, including male 17 cases, female 23 cases, with the age from 29 to 76 years (mean age 54 years). There were 11 patients with colon cancer and 29 with rectal cancer. The differentiations of the group were as follow: hige differentiated adenocarcinoma 23 cases, moderate differentiated adenocarcinoma 10 cases, low differentiated adenocarcinma 7 cases. Dukes stage: Dukes A9 cases, B12 cases, C12 cases, D7 cases. The status of lymph node metastasis: without lymph node metastasis 23 cases, with lymph node metastasis 17 cases. Another 10 patients with benign colon diseases were selected as control. Telomerase was extracted for TRAP. The kit for the detection of telomerase activity was bought from Huamei biological engineering company. The positive, negitive and blank control were taken from the kit, lysate and purified water, respectively. Products hybridization were conducted through Coated Mixcro-wells reverse hybridization method. hybridization products were tested by Model U2000 spectrophotometer to define its OD value at 450nm. The OD value of blank hybridization product is supposed to be zero. If the OD value of hybridization product in negative control is below 0.05, it is regarded as 0.05; if above 0.05, its actual value is counted. The OD value of the sample hybridization product is determined as telomerase activity positive if it is 2.1 times as or more than the negative control hybridization product. Results: In the 40 cases of colorectal carcinoma, 34 cases (85.0%) were found positive expression of telomerase activity. In the 10 cases of normal tissue, no any cases showed telomerase activity. There is significantly different between the two groups (P <0.05). No significant differences were found between the telomerase activity and the tumor location, differentiation, Dukes stage and lymph node metastasis. Conclusion: The activation of telomerase activity is common in colorectal cancer and may occur at early stage. The telomerase activity seems have very weak links with the prognosis in patients with colorectal cancer.更多还原