Fas在6种泌尿生殖系统肿瘤细胞系中的表达

Expression of Fas in Six Human Urogen ital Malignant Cell Lines

背景与目的:在肿瘤细胞中Fas蛋白的表达是研究治疗肿瘤及判断愈后的重要基础,本研究检测Fas在泌尿生殖系统恶性肿瘤细胞系中的表达情况。方法:采用流式细胞直接免疫荧光法、Westernblot和Northern杂交对6种泌尿生殖恶性肿瘤细胞系犤膀胱癌(T24,EJ,BIU-87)、肾癌(GRC-1,RCC-949)、前列腺癌(PC-3M)犦和一种原代培养正常肾间质成纤维细胞Fas分子的表达进行了检测。结果:直接免疫荧光流式细胞术检测细胞表面Fas在6种泌尿生殖恶性肿瘤细胞系均有表达,阳性率波动于16.51%-49.13%,但与原代培养的正常肾间质成纤维细胞(阳性率77.98%)相比,其阳性细胞百分率较低,细胞表面特异性免疫荧光强度较弱;6种泌尿生殖肿瘤细胞系Fas蛋白的表达均可用Westernblot法检测到一条分子量约48kDa的清晰条带,但表达强度不同,以BIU-87细胞最弱,而T24细胞最明显;只有在BIU-87细胞系中未能用Northern杂交方法检测到FasmRNA的表达,其它细胞系均有不同程度的表达,以PC-3M、T24细胞系较强,GRC-1、RCC-949细胞系次之,EJ细胞仅有微弱表达,正常肾间质成纤维细胞RFb表达较强的FasmRNA。结论:Fas广泛表达于泌尿生殖系恶性肿瘤培养细胞和正常细胞中,但在肿瘤中的表达情况明显低于正常细胞。

Background &Objective:Expression of Fas in tumors is an important basis for the treatment of the atients with tumor and determinatio n of the prognosis.This study was designed to investigate the expression of Fas in urogenital tumor cell lines.Methods:Using direct immunofluorescence fl ow cytometry,Western blot,we detec ted and Northern blot,the expression of Fas in six urinary malignant cell lines bladder carcinoma (T24,EJ,BIU-87),renal carcinoma (GRC-1,RCC-949),prostatic carcinoma (PC-3M) and one primary in vitro cultured ren al fibroblast.Results:Expression of Fas in 6urinary tumor c ell lines was commonly presented but with limited positive cell percenta ge(range16.51%-49.13%)and relatively lower fluorescence i ntensity,compared with expression of Fas in normal control of primary in vitro cultured renal fibr oblast(the positive rate of Fas reached 77.98%)measured by flow cytometry with dire ct immunofluorescence;In detection of expression of Fas in 6urogenital cell lines,a 48kDa clear band was found with the strongest intensity in T24cell line and the weakest in BIU-87.Fas mRNA from all tumor cell lines but BIU-87was detected.Conclusions:Fas was extensively expressed in uro genital malignant cells and normal cell.The lower expression of Fas in urinary malignant cell lines than th at in normal cells might be the reason for occurrence and progression of urinary malignant tumors.