DETECTION OF LAMININ RECEPTOR MESSENGER-RNA IN HUMAN CANCER CELL-LINES AND COLORECTAL TISSUES BY INSITU HYBRIDIZATION

被引:0
作者
CAMPO, E [1 ]
MONTEAGUDO, C [1 ]
CASTRONOVO, V [1 ]
CLAYSMITH, AP [1 ]
FERNANDEZ, PL [1 ]
SOBEL, ME [1 ]
机构
[1] NCI, PATHOL LAB, BLDG 10, RM 2A33, 9000 ROCKVILLE PIKE, BETHESDA, MD 20892 USA
关键词
D O I
暂无
中图分类号
R36 [病理学];
学科分类号
100104 ;
摘要
The 67-kd high-affinity laminin receptor (67 LR) is a gene product whose expression appears to be associated with the invasive and metastatic phenotype of a variety of human cancer cells. Northern blot hybridization bas been routinely used to quantify the level of 67 LR mRNA from total cellular RNA extracts homogenized tissue specimens or in vitro grown cell populations. This technique is useful to assess the average expression of the 67 LR mRNA of a particular sample but does not provide information about expression in specific cell types nor about heterogeneity of expression from cell to cell In this study, we analyzed the expression of 67 LR mRNA in four human cancer cell lines with varying degrees of expression 67 LR protein (renal cancer A-704, breast carcinoma MCF-7/4 and MCF-7/7, and pancreatic cancer Panc-1) using in situ hybridization performed with 67 LR riboprobes. Total cellular RNA was simultaneously extracted from the cell lines and hybridized on Northern blots with a 67 LR cDNA probe to assess the validity of the mRNA detection by in situ hybridization Sixty-seven LR mRNA expression was higher in Panc-1 and MCF-7/4 cells than in MCF-7/7 and renal carcinoma A-704. There was a direct correlation (R2 = 0.88) between the in situ hybridization analysis and the mRNA levels detected by Northern blot analysis. The in situ hybridization method showed a heterogeneous expression of the 67 LR mRNA in the four cell lines with different subpopulations of cells showing a range from negative to high levels of the message. Sixteen freshly frozen human colorectal tissues (seven adenocarcinomas, five matched normal mucosae, and four adenomas) were also analyzed by in situ hybridization The 67 LR mRNA was localized in normal and neoplastic epithelial cells. Adenocarcinoma cells showed a 1.6- to 5-fold higher expression (P < 0.02 according to the Wilcoxon-Mann-Whitney test) than did epithelial colonic cells from normal mucosae or adenomas. The signal tended to be stronger in poorly differentiated carcinomas and carcinomas with metastases than in moderately differentiated and nonmetastatic tumors. We conclude that the high expression of 67LR mRNA in colorectal tumors is due to an increased production by tumor cells. Furthermore, in situ hybridization is an effective method to detect the expression of LR mRNA in cultured cell lines as well as in frozen tissue sections.
引用
收藏
页码:1073 / 1083
页数:11
相关论文
共 46 条
[1]   INTEGRINS AND OTHER CELL-ADHESION MOLECULES [J].
ALBELDA, SM ;
BUCK, CA .
FASEB JOURNAL, 1990, 4 (11) :2868-2880
[2]  
ANGERER LM, 1987, IN SITU HYBRIDIZATIO, P42
[3]   LAMININ MOLECULAR DOMAINS WHICH ALTER METASTASIS IN A MURINE MODEL [J].
BARSKY, SH ;
RAO, CN ;
WILLIAMS, JE ;
LIOTTA, LA .
JOURNAL OF CLINICAL INVESTIGATION, 1984, 74 (03) :843-848
[4]   STRUCTURE AND FUNCTION OF LAMININ - ANATOMY OF A MULTIDOMAIN GLYCOPROTEIN [J].
BECK, K ;
HUNTER, I ;
ENGEL, J .
FASEB JOURNAL, 1990, 4 (02) :148-160
[5]  
BEVILACQUA G, 1989, CANCER RES, V49, P5185
[6]  
CASTRONOVO V, 1991, CANCER RES, V51, P5672
[7]  
CASTRONOVO V, 1991, J BIOL CHEM, V266, P20440
[8]   LAMININ AND FIBRONECTIN INCREASE THE STEADY-STATE LEVEL OF THE 67KD HIGH-AFFINITY METASTASIS-ASSOCIATED LAMININ RECEPTOR MESSENGER-RNA IN HUMAN CANCER-CELLS [J].
CASTRONOVO, V ;
SOBEL, ME .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1990, 168 (03) :1110-1117
[9]   BIOSYNTHESIS OF THE 67 KDA HIGH-AFFINITY LAMININ RECEPTOR [J].
CASTRONOVO, V ;
CLAYSMITH, AP ;
BARKER, KT ;
CIOCE, V ;
KRUTZSCH, HC ;
SOBEL, ME .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 177 (01) :177-183
[10]  
CASTRONOVO V, 1990, AM J PATHOL, V137, P1373