Frequent overexpression of CADM1/IGSF4 in lung adenocarcinoma
Section snippets
Materials and methods
Cells and mAbs. HepG2 [10], Caco-2 [11] and SKOv3 [11] were obtained from Riken Bioresource Center, ATCC and Dr. H. Shiku in Mie University School of medicine, respectively. mAbs used in this study were isolated in the previous study [4].
Flow cytometry (FCM). scFv-CL molecules fused with a truncated cp3 (scFv-CL-cp3) form of Ab was used for FCM analysis without purification. 5 × 105 cells were suspended in 500 μl to 1 ml of 2.5% normal goat serum/2.5% BSA/0.05% NaN3/PBS and stood for 30 min on ice.
Isolation of human mAbs against CADM1/IGSF4
As reported in a previous paper, 23 distinct anti CADM1 human mAbs have been isolated [4]. Table 1 summarized the results of screening showing how they had been isolated. Since the same clones were isolated from different screenings, 34 mAbs consisting of 23 distinct clones were listed in Table 1. Of those 34 mAbs 24 were isolated from screenings against hepatocarcinomas, five against colonic cancer, four against ovarian carcinoma and one against pancreatic carcinoma. Furthermore, 19 distinct
Discussion
In the previous work we identified 21 distinct Ags that are highly expressed on various carcinomas. Among them CADM1 that had been originally identified to be a tumor suppressor of human non-small cell lung cancer was included [4]. Many reports showed hypermethylation of the promoter region of CADM1 gene in various cancers including lung cancer, pancreatic carcinoma and breast cancer [13], [14]. Tumor suppressor activity mediated by CADM1/TSLC1 was shown by functional complementation experiment
Acknowledgments
This study was supported in part by a grant-in-aid for the 21st Century Center of Excellence (COE) Program of Fujita Health University from the Ministry of Education, Culture, Sports, Science, and Technology and by a grant from the New Energy and Industrial Technology Development Organization (NEDO) to Y.K.
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