Cloning of Novel Tumor Metastasis-Related Genes from the Highly Metastatic Human Lung Adenocarcinoma Cell Line Anip973

Fangli Liu; Yu Li; Yang Yu; Songbin Fu; Pu Li

doi:10.1016/S1673-8527(07)60020-4

Volume 34 Issue 3

Mar. 2007

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Article Navigation > Journal of Genetics and Genomics > 2007 > 34(3): 189-195

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Cloning of Novel Tumor Metastasis-Related Genes from the Highly Metastatic Human Lung Adenocarcinoma Cell Line Anip973

doi: 10.1016/S1673-8527(07)60020-4

a
Laboratory of Medical Genetics, Harbin Medical University, Harbin 150086, China
b
Department of Life Science and Engineering, Harbin Institute of Technology, Harbin 150001, China

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Corresponding author: E-mail address: lipu@ems.hrbmu.edu.cn (Pu Li)
Received Date: 2006-03-16
Accepted Date: 2006-06-02

Available Online: 2007-05-09

Publish Date: 2007-03-20

Abstract

Abstract

A cDNA library was successfully constructed from Anip973, a human lung adenocarcinoma cell line with high metastatic potential. NIH3T3 cells were stably transfected using this cDNA library and screened for morphological changes in a soft agar assay. Genomic DNA was isolated from putative clones and the integrated sequence was retrieved by PCR and sequencing. Three known genes, ribosomal protein L23, hypothetical protein FLJ22104, and serine protease inhibitor, kazal type 6 and a number of 5′-terminally truncated sequences were identified. Furthermore, cells transfected with ribosomal protein L23 was highly invasive compared with the empty vector as control (P < 0.02). These results indicate that the expression cloning of cDNA libraries in NIH3T3 cells and subsequent screening for loss of contact inhibition in soft agar is a viable tool for identifying tumor-related genes and ribosomal protein L23 gene plays a role in cell movement and metastasis.
- cDNA expression library,
- soft agar,
- oncogene,
- metastasis

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References(16)

References

[1]	Liang, P, Pardee, et al. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction Science, 257 (1992),pp. 967-971
[2]	Mahon, GM, Whitehead, et al. Retrovirus cDNA expression library screening for oncogenes Meth Enzymol, 332 (2001),pp. 211-221
[3]	Whitehead, I, Kirk, et al. Expression cloning of oncogenes by retroviral transfer of cDNA libraries Mol Cell Biol, 15 (1995),pp. 704-710
[4]	Tordaro, GJ, Green, et al. An assay for cellular transformation by SV40 Virology, 23 (1964),pp. 117-119
[5]	Barr, SM, Johnson, et al. Ras-induced colony formation and anchorage-independent growth inhibited by elevated expression of Puralpha in NIH3T3 cells J Cell Biochem, 81 (2001),pp. 621-638
[6]	Xia, P, Gamble, et al. An oncogenic role of sphingosine kinase Curr Biol, 10 (2000),pp. 1527-1530
[7]	Hoff, AO, Cote, et al. Neoplasia, 1 (1999),pp. 485-491
[8]	Fenrick, R, Wang, et al. TEL, a putative tumor suppressor, modulates cell growth and cell morphology of ras-transformed cells while repressing the transcription of stromelysin-1 Mol Cell Biol, 20 (2000),pp. 5828-5839
[9]	Wang, WR, Pan, et al. Screen metastatic human lung adenocarcinoma cell line in nude mice abdominal cavity and study of its biological character Chinese Journal of Oncology, 9 (1987),pp. 412-415
[10]	Daniel, L Retroviral ransfer of cDNA libraries to NIH3T3 cells; cloning novel oncogenes from Leukemic cell line HL-60 Am J Undergra Res, 4 (2002)
[11]	Jin, AW, Itahana, et al. Inhibition of HDM2 and activation of p53 by ribosomal protein L23 Mol Cell Biol, 24 (2004),pp. 7669-7680
[12]	Pestov, DG, Strezoska, et al. Evidence of p53-dependent cross-talk between ribosome biogenesis and the cell cycle: effects of nucleolar protein Bop1 on G(1)/S transition Mol Cell Biol, 21 (2001),pp. 4246-4255
[13]	Dai, MS, Zeng, et al. Ribosomal protein L23 activates p53 by inhibiting MDM2 function in response to ribosomal perturbation but not to translation inhibition Mol Cell Biol, 24 (2004),pp. 7654-7668
[14]	Rubbi, CP, Milner, et al. Disruption of the nucleolus mediates stabilization of p53 in response to DNA damage and other stresses EMBO J, 22 (2003),pp. 6068-6077
[15]	Colombo, E, Marine, et al. Mucleophosmin regulates the stability and transcriptional activity of p53 Nat Cell Biol, 4 (2002),pp. 529-533
[16]	Iapalucci-Espinoza, S, Franze-Fernandez, et al. Effect of proetein synthesis inhibitors and lox concentrations of actinomycin D on ribosomal RNA synthesis FEBS Lett, 107 (1979),pp. 281-284