Active c-erbB-2 induces short-term growth of FDC-P2 cells after IL-3 depletion

Wongsasant, Budsaba; Matsuda, Satoru; Yamamoto, Tadashi

doi:10.1016/0006-291x(91)92033-g

Cited by 2 publications

(1 citation statement)

References 35 publications

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“…The ErbB2(VE) gene is a mutant human ErbB2 (Genebank Accession # X 03363) with a substitution of valine 659 by glutamic acid within the transmembrane domain (Wongsasant et al, 1991). The ErbB2-B3CT gene contains the sequence coding for the carboxyl-terminal region of human ErbB3 (Genebank Accession # M 29366), amino acid 979-1342, combined with the sequence coding for the amino-terminal region of ErbB2(VE), amino acid 1-989, through a HindIII linker, which results in insertion of two amino acids, KL, between the two sequences.…”

Section: Plasmids Used In This Studymentioning

confidence: 99%

Activation of ErbB3–PI3-kinase pathway is correlated with malignant phenotypes of adenocarcinomas

et al. 2003

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Signet-ring cell carcinomas are malignant dedifferentiated carcinomas, which are frequently found in the stomach. We previously demonstrated that a 200 kDa protein is often constitutively phosphorylated on tyrosine and bound to phosphatidylinositol 3-kinase (PI3-kinase) in signetring cell carcinoma cells. In this study, we purified the 200 kDa protein from an extract of NUGC-4 cells, a cell line of signet-ring cell carcinoma, and identified it as ErbB3. ErbB3 was found to be phosphorylated selectively in dedifferentiated adenocarcinoma cell lines among various gastric cancer cell lines. Expression of a constitutively active chimeric receptor consisting of ErbB2 and ErbB3 in HCC2998 cells, a highly differentiated adenocarcinoma cell line, revealed that the signaling triggered by phosphorylation of ErbB3 was important for dedifferentiated phenotypes such as loss of cell-cell interaction and high expression of MUC1/DF3 antigen, a marker of the malignant tumors. Taken together, activation of ErbB3 pathway may contribute to the development of dedifferentiated carcinomas.

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Section: Plasmids Used In This Studymentioning

confidence: 99%

Activation of ErbB3–PI3-kinase pathway is correlated with malignant phenotypes of adenocarcinomas

et al. 2003

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High-efficiency identification of genes by functional analysis from a retroviral cDNA expression library

Wong

Chen

Chung

et al. 1994

J Virol

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Retroviral gene transfer efficiently delivers genes of interest stably into target cells, and expression cDNA cloning has been shown to be highly successful. Considering these two advantages, we now report a method by which one can identify genes stimulating cell growth through functional analysis. The first step requires the construction of a retroviral cDNA expression library and the optimization of transfection of vector DNA into virus packaging cells. The second step involves the cocultivation of target cells with libraries of retrovirusproducing cells, resulting in the amplification of target cells transduced with a gene(s) stimulating cell growth. Under standardized conditions of transfection, we detected an average of 4,000 independent clones per dish, among which expression of a retroviral 13-galactosidase gene at an abundance of 0.2% could be detected. Next, we demonstrated the augmentation of the sensitivity of the assay by retroviral infection and functional analysis. We did this by cocultivating factor-dependent (FD) cells with dishes of GP/E cells transfected with plasmids containing various molar ratios of pN2-IL3 DNA and retroviral library cDNA and by determining the highest dilution of pN2-1L3 which still resulted in the conversion of FD cells to factor independence. The retroviral interleukin-3 gene at an abundance as low as 0.001% could be detected. Indeed, we were able to detect from FD cells the development of factor-independent colonies with different phenotypes after retroviral transfer of cDNAs from an immortalized hemopoietic stem cell line. Thus, the combination of a standardized highefficiency DNA transfection and retrovirus-mediated gene transfer should facilitate the identification of genes capable of conferring to target FD cells a detectable new function or phenotype. By scaling up the size of the experiment realistically during screening, the assay can detect cDNA at an abundance of lower than 0.0001%.

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Active c-erbB-2 induces short-term growth of FDC-P2 cells after IL-3 depletion

Cited by 2 publications

References 35 publications

Activation of ErbB3–PI3-kinase pathway is correlated with malignant phenotypes of adenocarcinomas

Activation of ErbB3–PI3-kinase pathway is correlated with malignant phenotypes of adenocarcinomas

High-efficiency identification of genes by functional analysis from a retroviral cDNA expression library

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