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. 2003 Sep;9(9):2083-7.
doi: 10.3748/wjg.v9.i9.2083.

Gene transfer and expression of enhanced green fluorescent protein in variant HT-29c cells

Min Wang  1 , Lars Boenicke, Bradley D Howard, Ilka Vogel, Holger Kalthoff
Affiliations

Gene transfer and expression of enhanced green fluorescent protein in variant HT-29c cells

Min Wang et al. World J Gastroenterol. 2003 Sep.

Abstract

Aim: To study the expression of enhanced green fluorescent protein (EGFP) gene in retrovirally transduced variant HT-29 cells.

Methods: The retroviral vector prkat EGFP/neo was constructed and transfected into the 293T cell using a standard calcium phosphate precipitation method. HT-29c cells (selected from HT-29 cells) were transduced by a retroviral vector encoding the GEFP gene. The fluorescence intensity of colorectal carcinoma HT-29c cells after transduced with the EGFP bearing retrovirus was visualized using fluorescence microscope and fluorescence activated cell sorter (FACS) analysis. Multiple biological behaviors of transduced cells such as the proliferating potential and the expression of various antigens were comparatively analyzed between untransduced and transduced cells in vitro. EGFP expression of the fresh tumor tissue was assessed in vivo.

Results: After transduced, HT-29c cells displayed a stable and long-term EGFP expression under the nonselective conditions in vitro. After cells were successively cultured to passage 50 in vitro, EGFP expression was still at a high level. Their biological behaviors, such as expression of tumor antigens, proliferation rate and aggregation capability were not different compared to untransduced parental cells in vitro. In subcutaneous tumors, EGFP was stable and highly expressed.

Conclusion: An EGFP expressing retroviral vector was used to transduce HT-29c cells. The transduced cells show a stable and long-term EGFP expression in vitro and in vivo. These cells with EGFP are a valuable tool for in vivo research of tumor metastatic spread.

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Figures

Figure 1
Figure 1
Stable high level of EGFP expression of transduced HT-29c EGFP clone cells in vitro. × 100.
Figure 2
Figure 2
Stable high level of EGFP expressing s.c. tumor in nude rats formed from HT-29c EGFP clone#7 cells under fluo-rescence microscope (A: the tumor tissue was sliced at 0.8 mm, B: 60 μm cryosection). × 100.
Figure 3
Figure 3
Immunostaining of the s.c. tumor with mAb KL-1 (c) × 100.
See this image and copyright information in PMC

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References

    1. Prasher DC, Eckenrode VK, Ward WW, Prendergast FG, Cormier MJ. Primary structure of the Aequorea victoria green-fluorescent protein. Gene. 1992;111:229–233. - PubMed
    1. Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC. Green fluorescent protein as a marker for gene expression. Science. 1994;263:802–805. - PubMed
    1. Cubitt AB, Heim R, Adams SR, Boyd AE, Gross LA, Tsien RY. Understanding, improving and using green fluorescent proteins. Trends Biochem Sci. 1995;20:448–455. - PubMed
    1. Stearns T. Green fluorescent protein. The green revolution. Curr Biol. 1995;5:262–264. - PubMed
    1. Heim R, Cubitt AB, Tsien RY. Improved green fluorescence. Nature. 1995;373:663–664. - PubMed

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