Shao Li-na, Huang Qiong, Luo Min-jie, Lai Mao-de
Institute of Pathology and Forensic Medicine, Zhejiang University, Hangzhou 310031, China.
Zhonghua Bing Li Xue Za Zhi. 2003 Feb;32(1):14-9.
To get a complete cDNA sequence of B2 and evaluate the correlation on structure and expression between B2 and insulin like growth factor binding protein related protein 1 (IGFBP-rP1) in colorectal carcinomas, paired normal tissues, adenomas, tissues adjacent to the tumor, and colorectal carcinoma cell lines.
5'RACE (rapid amplification of cDNA end) was applied to get the sequence of the 5' end of B2. Semi-quantitative RT-PCR and immunohistochemistry were used to detect the expression of B2 in colorectal cancer tissues and cell lines (SW480, SW1116, SW620, HCT8, CoLo205 and LoVo).
A sequence of 1,125 bp was obtained by combining the sequence from 5'RACE product and the known sequence of B2. It shared 1,122/1,125 identities with IGFBP-rP1. At the level of mRNA, the expression of B2/IGFBP-rP1 was high in colorectal carcinomas, moderate in adenomas and tissues adjacent to tumor, low in normal tissues (P < 0.05). Five cell lines except SW480 showed no expression of B2/IGFBP-rP1. A significant difference was obtained in the immunoreactivity of B2/IGFBP-rP1 between normal tissue and cancer (P < 0.05). In 28.9% (22/76) samples, cancer cells locating at the invasive front of cancer nest had a stronger staining of B2/IGFBP-rP1 than those surrounding the lumen. These samples had also an increased frequency of lymph node metastases, increased depth of invasion and a stronger staining of B2/IGFBP-rP1 than in other samples (P < 0.05).
B2 is the same gene as IGFBP-rP1. Overexpression of B2/IGFBP-rP1 may play an important role in the initiation and promotion of colorectal cancer. Its overexpression in invading tumor cells may be linking with an increased potential of invasion.
获取B2的完整cDNA序列,并评估B2与胰岛素样生长因子结合蛋白相关蛋白1(IGFBP-rP1)在结直肠癌、配对的正常组织、腺瘤、肿瘤旁组织及结直肠癌细胞系中的结构和表达相关性。
应用5'RACE(cDNA末端快速扩增)获取B2的5'端序列。采用半定量RT-PCR和免疫组织化学检测B2在结直肠癌组织及细胞系(SW480、SW1116、SW620、HCT8、CoLo205和LoVo)中的表达。
将5'RACE产物序列与已知的B2序列相结合,获得了一个1125bp的序列。它与IGFBP-rP1有1122/1125的同源性。在mRNA水平,B2/IGFBP-rP1在结直肠癌中表达高,在腺瘤和肿瘤旁组织中表达中等,在正常组织中表达低(P<0.05)。除SW480外的5种细胞系均未检测到B2/IGFBP-rP1的表达。正常组织与癌组织中B2/IGFBP-rP1的免疫反应性有显著差异(P<0.05)。在28.9%(22/76)的样本中,位于癌巢浸润前沿的癌细胞B2/IGFBP-rP1染色强于围绕管腔的癌细胞。这些样本的淋巴结转移频率增加、浸润深度增加,且B2/IGFBP-rP1染色强于其他样本(P<0.05)。
B2与IGFBP-rP1为同一基因。B2/IGFBP-rP1的过表达可能在结直肠癌的发生和发展中起重要作用。其在侵袭性肿瘤细胞中的过表达可能与侵袭潜能增加有关。