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Overexpression, purification, molecular characterization and the effect on tumor growth of ribosomal protein L22 from the Giant Panda (Ailuropoda melanoleuca)

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Abstract

The ribosomal protein L22 (RPL22) protein belongs to the L22E family of ribosomal proteins. It is located in the cytoplasm. The purpose of this paper was to explore the structure and anti-cancer function of RPL22 of the Giant Panda (Ailuropoda melanoleuca). The cDNA of RPL22 was cloned successfully from the Giant Panda using RT-PCR technology. We constructed a recombinant expression vector containing RPL22 cDNA and over-expressed it in Escherichia coli using pET28a plasmids. The expression product obtained was purified by using Ni chelating affinity chromatography. The result indicated that the length of the fragment cloned is 414 bp, and it contains an open-reading frame of 387 bp encoding 128 amino acids. Primary structure analysis revealed that the molecular weight of the putative RPL22 protein is 14.74 kDa with a theoretical pI 9.21. The RPL22 gene can be really expressed in E. coli and the RPL22 protein, fusioned with the N-terminally His-tagged protein, gave rise to the accumulation of an expected 20.1 kDa polypeptide. The data showed that the recombinant protein RPL22 had a time- and dose-dependency on the cell growth inhibition rate. The human laryngeal carcinoma Hep-2 cells treated with 0.05–6 μg/ml of RPL22 for 24 h displayed significant cell growth inhibition (p < 0.05, n = 8) in assayed using MTT compared to the control (untreated) cells. The data indicate that the effect at low concentrations is better than high concentrations, and the concentration of 1.5 μg/ml has the best rate of growth inhibition of 47.70 %. The inhibitory rate in mice treated with 1.5 μg/ml RPL22 protein can reach 43.75 %. Histology of tumor organs shows that the tissues arranged looser in RPL22 group than those in control group. Meanwhile, there is no obvious damage to other organs, such as heart, lung and kidney. Further research is on going to determine the bioactive principle(s) of recombinant protein RPL22 responsible for its anticancer activity.

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References

  1. Wool IG, Chan YL, Glück A (1996) Structure and evolution of mammalian ribosomal proteins. Biochem Cell Biol 73:933–947

    Article  Google Scholar 

  2. Wool IG (1996) Extraribosomal functions of ribosomal proteins. Trend Biochem Sci 21:164–165

    Article  CAS  PubMed  Google Scholar 

  3. Fabijanski S, Pellegrini MM (1981) Identification of proteins at the peptidyl-tRNA binding site of rat liver ribosomes. Mole Gen Genet MGG 184:551–556

    Article  CAS  Google Scholar 

  4. Nissen P, Hansen J, Ban N, Moore PB, Steitz TA (2000) The structural basis of ribosome activity in peptide bond synthesis. Science 289:920–930

    Article  CAS  PubMed  Google Scholar 

  5. Wang Q, Yang C, Zhou J, Wang X, Wu M, Liu Z (2001) Cloning and characterization of full-length human ribosomal protein L15 cDNA which was overexpressed in esophageal cancer. Gene 263:205–209

    Article  CAS  PubMed  Google Scholar 

  6. Kim JH, You KR, Kim IH, Cho BH, Kim CY, Kim DG (2004) Over-expression of the ribosomal protein L36a gene is associated with cellular proliferation in hepatocellular carcinoma. Hepatology 39:129–138

    Article  CAS  PubMed  Google Scholar 

  7. Lopez CD, Martinovsky G, Naumovski L (2002) Inhibition of cell death by ribosomal protein L35a. Cancer Lett 180:195–202

    Article  CAS  PubMed  Google Scholar 

  8. Yang F, Liu WP (2005) Study on the relation of ribosomal protein gene and human diseases. Chin J Clin Exp Pathol 3:354–356

    Google Scholar 

  9. Klein DJ, Moore PB, Steitz TA (2004) The roles of ribosomal proteins in the structure assembly, and evolution of the large ribosomal subunit. J Mol Biol 340:141–177

    Article  CAS  PubMed  Google Scholar 

  10. Liao MJ, Zhu MY, Zhang ZH, Zhang AJ (2003) cDNA cloning of growth hormone from giant panda (Ailuropoda melanoleuca) and its expression in Escherichia coli. Comp Biochem Phys B 135:109–116

    Google Scholar 

  11. Du YJ, Luo XY, Hao YZ, Zhang T, Hou WR (2007) Cloning and overexpression of acidic ribosomal phosphoprotein P1 gene (RPLP1) from the Giant Panda. Int J Biol Sci 3:428–433

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Hou WR, Du YJ, Chen Y, Wu X, Peng ZS, Yang J, Zhou CQ (2007) Nucleotide sequence of cDNA encoding the mitochondrial precursor protein of the ATPase inhibitor from the Giant Panda (Ailuropoda melanoleuca). DNA Cell Biol 26:799–802

    Article  CAS  PubMed  Google Scholar 

  13. Du YJ, Hou WR, Peng ZS, Zhou CQ (2008) cDNA cloning and sequences analysis of acidic ribosomal phosphoprotein P1 (RPLP1) from Giant Panda. Acta Theriologica Sinica 28:75–80

    Google Scholar 

  14. Wu GF, Hou YL, Hou WR, Song Y, Zhang T (2010) Giant panda ribosomal protein S14: cDNA, genomic sequence cloning, sequence analysis, and overexpression. Genet Mol Res 9(4):2004–2015

    Article  CAS  PubMed  Google Scholar 

  15. Hou WR, Luo XY, Du YJ, Chen Y, Wu X, Peng ZS, Yang J, Zhou CQ (2008) cDNA cloning and sequences analysis of RPS15 from the Giant Panda. Recent Pat DNA Seq 2:16–19

    Article  CAS  Google Scholar 

  16. Hou YL, Hou WR, Ren ZL, Hao YZ, Zhang T (2009) cDNA cloning and overexpression of ribosomal protein S19 gene (RPS19) from the Giant Panda. Cell Biol 1:1–47

    Google Scholar 

  17. Sun B, Hou YL, Hou WR, Zhang SN, Ding X, Su XL (2012) cDNA cloning, overexpression, purification and pharmacologic evaluation for anticancer activity of ribosomal protein L23A Gene (RPL23A) from the Giant Panda. Int J Mol Sci 13:2133–2147

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Nutting CM, Robinson M, Birchall M (2008) Survival from laryngeal cancer in England and Wales up to 2001. Br J Cancer 99:38–39

    Article  Google Scholar 

  19. Zhou J, Callapina M, Goodall GJ, Brune B (2004) Functional integrity of nuclear factor kappaB, phosphatidylinositol 30-kinase, and mitogen-activated protein kinase signaling allows tumor necrosis factor alpha-evoked Bcl-2 expression to provoke internal ribosome entry site-dependent translation of hypoxiainducible factor 1 alpha. Cancer Res 64:9041–9048

    Article  CAS  PubMed  Google Scholar 

  20. Lai MD, Xu J (2007) Ribosomal proteins and colorectal cancer. Curr Genomics 8:43–49

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  21. Cheng Q, Lau WM, Chew SH, Ho TH, Tay SK, Hui KM (2002) Identification of molecular markers for the early detection of human squamous cell carcinoma of the uterine cervix. Br J Cancer 86:274–281

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Henry JL, Coggin DL, King CR (1993) High-level expression of the ribosomal protein L19 in human breast tumors that overexpress erbB-2. Cancer Res 15:1403–1408

    Google Scholar 

  23. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D (2011) Global cancer statistics. CA Cancer J Clin 61(2):69–90

    Article  PubMed  Google Scholar 

  24. Tolar J, Neglia JP (2003) Transplacental and other routes of cancer transmission between individuals. J Pediatr Hematol Oncol 25(6):430–434

    Article  PubMed  Google Scholar 

  25. Yu L, Li HZ, Lu SM, Liu WW, Li JF, Wang HB, Xu W (2009) Alteration in TWIST expression: possible role in paclitaxel-induced apoptosis in human laryngeal carcinoma Hep-2 cell line. Croat Med J 50:536–542

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  26. Chiao PJ, Shin DM, Sacks PG, Hong WK, Tainsky MA (1992) Elevated expression of the ribosomal protein S2 gene in human tumors. Mol Carcinog 5:219–231

    Article  CAS  PubMed  Google Scholar 

  27. Barnard GF, Staniunas RJ, Bao S et al (1992) Increased expression of human ribosomal phosphoprotein P0 messenger RNA in hepatocellular carcinoma and colon carcinoma. Cancer Res 52:3067–3072

    CAS  PubMed  Google Scholar 

  28. Deni MG, Chadeneau C, Lecabellec MT et al (1993) Overexpression of the S13 ribosomal protein in actively growing cells. Int J Cancer 55:275–280

    Article  Google Scholar 

  29. Ropolo M, Geroldi A, Rossi O, Degan P, Zupo S, Poggi A, Frosina G (2004) Expression of the Drosophila melanogaster S3 ribosomal/repair protein in T24 human bladder cells. Anticancer Res 24:3811–3818

    CAS  PubMed  Google Scholar 

  30. Clark DE, Errington TM, Smith JA, Frierson HF, Weber MJ, Lannigan DA (2005) The serine/threonine protein kinase, p90 ribosomal S6 kinase, is an important regulator of prostate cancer cell proliferation. Cancer Res 65:3108–3116

    CAS  PubMed  Google Scholar 

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Acknowledgments

This project was supported by Application Foundation Project of Sichuan Province (2013JY0094), National Natural Science Foundation of China (31200012) and Doctor Startup Foundation Project of China West Normal University (11B019) and (11B020).

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Authors and Affiliations

  1. Key Laboratory of Southwest China Wildlife Resources Conservation (Ministry of Education), College of Life Sciences, China West Normal University, 1# Shida Road, Nanchong, 637009, Sichuan, China

    Jian Li, Yiling Hou, Xiang Ding, Wanru Hou, Bo Song & Yichun Zeng

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  1. Jian Li
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  2. Yiling Hou
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  3. Xiang Ding
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  4. Wanru Hou
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  5. Bo Song
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Correspondence to Xiang Ding.

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Jian Li and Yiling Hou are co-first authors.

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Li, J., Hou, Y., Ding, X. et al. Overexpression, purification, molecular characterization and the effect on tumor growth of ribosomal protein L22 from the Giant Panda (Ailuropoda melanoleuca). Mol Biol Rep 41, 3529–3539 (2014). https://doi.org/10.1007/s11033-014-3217-3

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  • DOI: https://doi.org/10.1007/s11033-014-3217-3

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