Functional expression of recombinant canstatin in stably transformed Drosophila melanogaster S2 cells

Lee, Jong Min; Jeon, Hoon; Sohn, Bong Hee; Chung, In Sik

doi:10.1016/j.pep.2006.11.016

Cited by 13 publications

(15 citation statements)

References 15 publications

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“…17 Recombinant canstatin from stably transformed Drosophila S2 cells was purified to homogeneity using a simple one-step Ni-NTA affinity fractionation, as described previously. 17 …”

Section: Preparation Of Purified Recombinant Canstatinmentioning

confidence: 99%

Recombinant canstatin inhibits angiopoietin‐1‐induced angiogenesis and lymphangiogenesis

Jeon

Yoo

Park

et al. 2011

Intl Journal of Cancer

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We describe the effect of recombinant canstatin, the NC1 domain of the a2 chain of Type IV collagen, on suppression of angiogenesis and lymphangiogenesis both in vitro and in vivo. Recombinant canstatin produced from stably transformed Drosophila S2 cells reduced the expression of angiopoietin-1 in hypoxia mimetic agent, CoCl 2 -treated CT-26 cells. Recombinant canstatin inhibited proliferation, tube formation and migration of human angiopoietin-1 (rhAngpt-1)-treated human umbilical vein endothelial cells (HUVEC) and lymphatic endothelial cells (LEC). Recombinant canstatin suppressed the expression of Tie-2 and vascular endothelial growth factor-3 (VEGFR-3) transcripts in rhAngpt-1-treated HUVEC and LEC, respectively. The inhibitory effect of recombinant canstatin on tumor growth was also investigated using a heterotopic CT-26 colon carcinoma animal (BALB/c mice) model. Recombinant canstatin reduced the final volume and weight of tumors, and blood and lymphatic vessel densities of tumors, which were evaluated by CD-31 and LYVE-1 immunostaining. Immunohistochemical analysis showed that recombinant canstatin dramatically reduced the expression of angiopoietin-1 in CT-26 colon carcinoma-induced tumor, but not the expression of VEGF-C. Tie-2 and VEGFR-3 expressions were also reduced in recombinant canstatin-treated tumors. These results indicate that recombinant canstatin has anti-tumoral activities against CT-26 colon carcinoma cells. Recombinant canstatin reduces the expression of angiopoietin-1 in hypoxia-induced CT-26 cells and inhibits the angiogenic and lymphangiogenic signaling induced by angiopoietin-1. Recombinant canstatin probably inhibits angiogenesis and lymphangiogenesis via suppression of the integrin-dependent FAK signaling induced by angiopoietin-1/Tie-2 and/or VEGFR-3.

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“…17 Recombinant canstatin from stably transformed Drosophila S2 cells was purified to homogeneity using a simple one-step Ni-NTA affinity fractionation, as described previously. 17 …”

Section: Preparation Of Purified Recombinant Canstatinmentioning

confidence: 99%

Recombinant canstatin inhibits angiopoietin‐1‐induced angiogenesis and lymphangiogenesis

Jeon

Yoo

Park

et al. 2011

Intl Journal of Cancer

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“…Recombinant human canstatin proteins were obtained by transformation of Drosophila melanogaster S2 cells, transfected with a plasmid containing canstatin cDNA using the lipofectamine method (Lee et al 2007). Recombinant canstatin from stably transfected Drosophila S2 cells was purified to homogeneity using a simple one-step Ni NTA affinity fractionation (Lee et al 2007).…”

Section: Preparation Of Purified Canstatinmentioning

confidence: 99%

“…We have described earlier the stable expression of the cDNA for human canstatin in stably transfected Drosophila S2 cells and purification of recombinant canstaitn using metal chelate affinity fractionation (Lee et al 2007). Recently, we established an orthotopic AT-84 oral squmamous cell carcinoma animal (C3H/He) model and examined in vivo inhibitory effects of tumstatin on tumor growth in this model (Chung et al 2008).…”

Section: Introductionmentioning

confidence: 99%

Recombinant canstatin inhibits tumor growth in an orthotopic AT-84 oral squamous cell carcinoma model

et al. 2009

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The inhibitory effect of recombinant canstatin on tumor growth was investigated using an orthotopic oral squamous cell carcinoma (AT-84 cells) animal (C3H/HeN) model. Recombinant canstatin from stably transfected Drosophila S2 cells was purified to homogeneity using a simple one-step Ni NTA affinity fractionation. In our oral cancer model, the final volume and weight of tumors in groups treated with purified canstatin were both reduced to 44% of values for a control group treated with PBS. Blood or lymphatic vessel densities of tumors in the canstatin-treated group were reduced to 72% and 44% of control group values, respectively. Recombinant canstatin at 20 microg/ml effectively inhibited tube formation in HUVEC and lymphatic endothelial cells. Our results show that recombinant canstatin has anti-tumoral effects against primary oral squamous cell carcinoma.

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“…The two most utilized promoters are the constitutive actin promoter and the inducible metallothionein promoter which is activated by the addition of heavy metal in the culture medium (Chung and Keller 1990a, b). Several complex glycoproteins were already expressed in the S2 cell system, using these promoters (Mallender et al 2001;Zhang et al 2007;Scotter et al 2006;Brillet et al 2006;Kim et al 2005;Johansson et al 2007;Jennings et al 2006;Li et al 2005;Lim et al 2004;Lee et al 2007). Aiming the expression of high levels of RVGP under the control of these promoters in the S2 cell system, for vaccination as well as structure/function evaluation, many studies were already carried out on cell growth and heterologous recombinant protein expression kinetics (Yokomizo et al 2007;Galesi et al 2008;Swiech et al 2008a;Batista et al 2009;Ventini et al 2010;Lemos et al 2009), as well as on metabolism and synthesis of secondary products (Swiech et al 2008b, c) and culture medium formulation and supplementation (Galesi et al 2007;Batista et al 2008Batista et al , 2011Mendonça et al 2008Mendonça et al , 2009.…”

Section: Introductionmentioning

confidence: 99%

Kinetic studies of recombinant rabies virus glycoprotein (RVGP) cDNA transcription and mRNA translation in Drosophila melanogaster S2 cell populations

et al. 2013

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Recombinant rabies virus glycoprotein (RVGP) was expressed in cell membranes of stably transfected Drosophila S2 cells using constitutive and inducible promoters. Although with quantitative differences of RVGP expression in both systems, the cDNA transcription, as evaluated by relative RVGP mRNA levels measured by qRT-PCR, sustained the amount of RVGP producing cells and the RVGP volumetric (P RVGP ) productivity. At the transition to the stationary cell growth phase, once the cell culture slowed down its rate of multiplication, an accumulation of RVGP mRNA and RVGP was clearly observed in both cell populations. Nevertheless, cell cultures performed under sub-optimal temperatures indicated that an envisaged increase in the RVGP production is not only dependent on cell growth rate, but essentially on optimal cell metabolic state.

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Functional expression of recombinant canstatin in stably transformed Drosophila melanogaster S2 cells

Cited by 13 publications

References 15 publications

Recombinant canstatin inhibits angiopoietin‐1‐induced angiogenesis and lymphangiogenesis

Recombinant canstatin inhibits angiopoietin‐1‐induced angiogenesis and lymphangiogenesis

Recombinant canstatin inhibits tumor growth in an orthotopic AT-84 oral squamous cell carcinoma model

Kinetic studies of recombinant rabies virus glycoprotein (RVGP) cDNA transcription and mRNA translation in Drosophila melanogaster S2 cell populations

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