Abstract
Tubulin polymerization promoting protein 3 (TPPP3), a member of the TPPP family, can induce tubulin polymerization and microtubule bundling. Previously, it has been shown that TPPP3 plays an important role in cell proliferation. Depletion of TPPP3 by microRNA-based RNA interference (RNAi) inhibits cell growth, arrests cell cycles, and causes mitotic abnormalities in HeLa cells. In the present study, we knocked down TPPP3 in Lewis lung carcinoma (LLC) cells with the same RNAi construct, and observed a retarded tumor cell growth in vitro. Furthermore, C57BL/6 mice that received subcutaneously injected LLC cells in which TPPP3 was knocked down showed a pronounced reduction in tumor progression. The migration/invasion activity of TPPP3-knockdown LLC cells was significantly suppressed in a transwell chamber migration assay. When these cells were injected into the tail veins of C57BL/6 mice, they exhibited milder lung metastasis compared with control tumor cells. Taken together, these findings suggested that the TPPP3 gene played an important role in tumorigenesis and metastasis, and it could potentially become a novel target for cancer therapy.
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References
Desai A, Mitchison TJ (1997) Microtubule polymerization dynamics. Annu Rev Cell Dev Biol 13:83–117
MacRae TH (1992) Towards an understanding of microtubule function and cell organization: an overview. Biochem Cell Biol 70:835–841
Sakai H (1994) Microtubules in mitosis. Cell Struct Funct 19:57–62
Gadde S, Heald R (2004) Mechanisms and molecules of the mitotic spindle. Curr Biol 14:R797–R805
Zhu H, Fang K, Fang G (2009) Mechanism, function and regulation of microtubule-dependent microtubule amplification in mitosis. Mol Cells 27:1–3
Mandelkow E, Mandelkow EM (1995) Microtubules and microtubule-associated proteins. Curr Opin Cell Biol 7:72–81
Seki N, Hattori A, Sugano S, Suzuki Y, Nakagawara A, Muramatsu M, Hori T, Saito T (1999) A novel human gene whose product shares significant homology with the bovine brain-specific protein p25 on chromosome 5p15.3. J Hum Genet 44:121–122
Zhang Z, Wu C, Huang W, Wang S, Zhao E, Huang Q, Xie Y, Mao Y (2002) A novel human gene whose product shares homology with bovine brain-specific protein p25 is expressed in fetal brain but not in adult brain. J Hum Genet 47:266–268
Hlavanda E, Kovács J, Oláh J, Orosz F, Medzihradszky KF, Ovádi J (2002) Brain-specific p25 protein binds to tubulin and microtubules and induces aberrant microtubule assemblies at substoichiometric concentrations. Biochemistry 41:8657–8664
Vincze O, Tökési N, Oláh J, Hlavanda E, Zotter A, Horváth I, Lehotzky A, Tirián L, Medzihradszky KF, Kovács J, Orosz F, Ovádi J (2006) Tubulin polymerization promoting proteins (TPPPs): members of a new family with distinct structures and functions. Biochemistry 45:13818–13826
Zhou W, Wang X, Li L, Feng X, Yang Z, Zhang W, Hu R (2010) Depletion of tubulin polymerization promoting protein family member 3 suppresses HeLa cell proliferation. Mol Cell Biochem 333:91–98
Wehrle-Haller B, Imhof BA (2003) Actin, microtubules and focal adhesion dynamics during cell migration. Int J Biochem Cell Biol 35:39–50
Etienne-Manneville S (2004) Actin and microtubules in cell motility: which one is in control? Traffic 5:470–477
Waterman-Storer CM, Salmon E (1999) Positive feedback interactions between microtubule and actin dynamics during cell motility. Curr Opin Cell Biol 11:61–67
Acevedo K, Li R, Soo P, Suryadinata R, Sarcevic B, Valova VA, Graham ME, Robinson PJ, Bernard O (2007) The phosphorylation of p25/TPPP by LIM kinase 1 inhibits its ability to assemble microtubules. Exp Cell Res 313:4091–4106
Vasiliev JM (2004) Cytoskeletal mechanisms responsible for invasive migration of neoplastic cells. Int J Dev Biol 48:425–439
Suresh S (2007) Biomechanics and biophysics of cancer cells. Acta Biomater 3:413–438
Lehotzky A, Tirián L, Tökési N, Lénárt P, Szabó B, Kovács J, Ovádi J (2004) Dynamic targeting of microtubules by TPPP/p25 affects cell survival. J Cell Sci 117:6249–6259
Hlavanda E, Klement E, Kókai E, Kovács J, Vincze O, Tökési N, Orosz F, Medzihradszky KF, Dombrádi V, Ovádi J (2007) Phosphorylation blocks the activity of tubulin polymerization-promoting protein (TPPP): identification of sites targeted by different kinases. J Biol Chem 282:29531–29539
Orosz F (2009) Apicortin, a unique protein, with a putative cytoskeletal role, shared only by apicomplexan parasites and the placozoan Trichoplax adhaerens. Infect Genet Evol 9:1275–1286
Watanabe T, Noritake J, Kaibuchi K (2005) Regulation of microtubules in cell migration. Trends Cell Biol 15:76–83
Yamaguchi H, Wyckoff J, Condeelis J (2005) Cell migration in tumors. Curr Opin Cell Biol 17:559–564
Acknowledgments
This work was supported by grants to Prof. RM Hu from National Natural Science Foundation of China (Grant number: 30670999 and 30770854), Shanghai Science and Technology Commission (08dj1400605, 08JC1403200), and National Key Basic Research and Development Program (Grant number: 2002CB713703).
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Zhou, W., Li, J., Wang, X. et al. Stable knockdown of TPPP3 by RNA interference in Lewis lung carcinoma cell inhibits tumor growth and metastasis. Mol Cell Biochem 343, 231–238 (2010). https://doi.org/10.1007/s11010-010-0518-2
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DOI: https://doi.org/10.1007/s11010-010-0518-2