Hedgehog信号通路转录因子Gli1蛋白在结直肠癌组织中的表达

Expression of Hedgehog Signal Path Transcription Factor Gli1 in the Human Colorectal Carcinoma

目的探讨Hedgehog信号通路转录因子脑胶质瘤相关癌基因1(Gli1)蛋白在结直肠癌组织中的表达及作用。方法免疫组化法检测Gli1蛋白在147例手术切除结直肠癌组织中的表达;Western blotting法检测Gli1蛋白在19例结直肠癌新鲜组织和2个结肠癌细胞系中的表达;同时用共聚焦激光显微镜观察Gli1转录水平的抑制剂GANT61对8例直肠癌组织中直接分离的肿瘤细胞的影响。结果 147例结直肠癌根治切除标本中,Gli1蛋白在78.2%(115/147)的结直肠癌组织中表达升高,阳性信号表达在肿瘤细胞的细胞核和细胞浆。Gli1蛋白在结直肠癌组织中的表达与结直肠癌浸润前缘肿瘤细胞团数目、浸润深度、淋巴结转移、TNM分期有关(P<0.05)。在5例Gli1蛋白阳性的结直肠癌组织中,间质细胞,包括肿瘤相关纤维母细胞和内皮细胞内高表达Gli1蛋白。Gli1蛋白在结直肠癌组织和2个结肠癌细胞系中的表达也升高。对结直肠癌组织中直接分离出的肿瘤细胞,培养以后加入GANT61,可以明显抑制肿瘤细胞内Gli1蛋白的表达,而对肿瘤相关纤维母细胞没有显著影响。结论 Gli1蛋白的活化可能增强肿瘤细胞的侵袭和转移能力,从而促进结直肠癌的进展;GANT61可能为进展期结直肠癌的治疗提供新的契机。

Objective To explore the expression and significance of Hedgehog signal path transcription factor glioma associated oncogene 1（ Gli1） in the human colorectal carcinoma. Methods Expressions of Gli1 protein was detected and evaluated in the 147 cases of human colorectal carcinoma by immunohistochemistry. Western blotting was used to analyze Gli1 protein in the fresh colorectal carcinoma tissues and the two cell lines of colon carcinoma. Additionally,the carcinoma cells directly isolated from 8 cases of colorectal carcinoma tissues were added with GANT61,and were observed by confocal laser microscopy. Results Gli1 protein was detected in the nucleus and cytoplasm of carcinoma cells in the 78. 2%（ 115 /147） of the 147 cases of human colorectal carcinoma. Statistically,the expression of Gli1 protein in the human colorectal carcinoma was significantly correlated with the tumor cell clusters of invasion edge,deepness of tumor invasion,lymph node metastasis,and TNM staging,respectively（ P 〈0. 05）. In 5 cases of human colorectal carcinoma with positive expression of Gli1 protein,Gli1 protein was also detected in the tumor stroma cells,including cancer associated fibroblasts and endothelial cells. Gli1 protein high expression was confirmed by Western blotting in the carcinoma tissues and the two cell lines. In addition,GANT61 inhibited the expression of Gli1 protein in the carcinoma cells isolated from the CRC tissues significantly,but not for the carcinomaassociated fibroblasts. Conclusion Overexpression of Gli1 protein may involve in the progression of human colorectal carcinoma through increasing tumor invasion and metastasis; furthermore,GANT61 may be a new drug for the therapy of advanced CRC.