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HIV-1 Nef and KSHV oncogene K1 synergistically promote angiogenesis by inducing cellular miR-718 to regulate the PTEN/AKT/mTOR signaling pathway.

Xue M, Yao S, Hu M, Li W, Hao T, Zhou F, Zhu X, Lu H, Qin D, Yan Q, Zhu J, Gao SJ, Lu C - Nucleic Acids Res. (2014)

Bottom Line: Kaposi's sarcoma (KS) is an AIDS-defining cancer with aberrant neovascularization caused by KS-associated herpesvirus (KSHV).Furthermore, Nef and K1 induced cellular miR-718, which inhibited PTEN expression by directly targeting a seed sequence in the 3' UTR of its mRNA.Our results demonstrate an essential role of miR-718/AKT/mTOR axis in AIDS-KS and thus may represent an attractive therapeutic target.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, P.R. China Key Laboratory of Pathogen Biology of Jiangsu Province, Nanjing Medical University, Nanjing, P.R. China Department of Microbiology, Nanjing Medical University, Nanjing 210029, P.R. China Department of Physiology, Xuzhou Medical College, Xuzhou, Jiangsu 221004, P.R. China.

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Nef promotes K1-induced angiogenesis and tumorigenesis in the CAM and nude mice models. (A) Nef promoted K1-induced angiogenesis in HUVECs. HUVECs transduced with K1, incubated with soluble Nef protein for 72 h or both were mixed with Matrigel and subsequently implanted onto the CAM. Representative photographs of angiogenesis on the CAM are shown. (B) Quantification of results in (A). The number of blood vessels was normalized to that of Matrigel alone. Data represent mean ± SD from three independent experiments (n = 3), each experiment containing 10 technical replicates. (C) Nef enhanced K1-induced angiogenesis in EA.hy926 cells. EA.hy926 cells transduced with K1, Nef or both were mixed with Matrigel and subsequently implanted onto the CAM. The number of blood vessels was normalized to that of Matrigel alone. Data represent mean ± SD from three independent experiments (n = 3), each experiment containing 10 technical replicates. (D) Hematoxylin and eosin staining analysis of histologic features (top; original magnification, ×100) and immunohistochemical staining analysis of the expression of SMA and VEGF (middle and bottom; original magnification, ×200) in tumor tissues from the CAMs induced by EA.hy926 cells transduced with K1 and Nef. Black arrows point to neovascularization and hemorrhagic foci. (E) Quantification of results in (D). (F) Western blot analysis of total PTEN and phosphorylation levels of PI3K, AKT and mTOR in tumor tissues from the CAMs treated as in (D). (G) Nef enhanced K1-induced angiogenesis in nude mice. EA.hy926 cells transduced by K1, Nef or both were examined for their proangiogenic effects in Matrigel plug assay in nude mice as described in the ‘Materials and Methods’ section. Representative photographs of angiogenesis in the nude mice are shown. (H) The hemoglobin level of the Matrigel plugs treated as in (G) was determined with O.D. value at 540 nm. Data represent mean ± SD, each group with five tumors (n = 5). Three independent experiments were performed and similar results were obtained.
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Figure 2: Nef promotes K1-induced angiogenesis and tumorigenesis in the CAM and nude mice models. (A) Nef promoted K1-induced angiogenesis in HUVECs. HUVECs transduced with K1, incubated with soluble Nef protein for 72 h or both were mixed with Matrigel and subsequently implanted onto the CAM. Representative photographs of angiogenesis on the CAM are shown. (B) Quantification of results in (A). The number of blood vessels was normalized to that of Matrigel alone. Data represent mean ± SD from three independent experiments (n = 3), each experiment containing 10 technical replicates. (C) Nef enhanced K1-induced angiogenesis in EA.hy926 cells. EA.hy926 cells transduced with K1, Nef or both were mixed with Matrigel and subsequently implanted onto the CAM. The number of blood vessels was normalized to that of Matrigel alone. Data represent mean ± SD from three independent experiments (n = 3), each experiment containing 10 technical replicates. (D) Hematoxylin and eosin staining analysis of histologic features (top; original magnification, ×100) and immunohistochemical staining analysis of the expression of SMA and VEGF (middle and bottom; original magnification, ×200) in tumor tissues from the CAMs induced by EA.hy926 cells transduced with K1 and Nef. Black arrows point to neovascularization and hemorrhagic foci. (E) Quantification of results in (D). (F) Western blot analysis of total PTEN and phosphorylation levels of PI3K, AKT and mTOR in tumor tissues from the CAMs treated as in (D). (G) Nef enhanced K1-induced angiogenesis in nude mice. EA.hy926 cells transduced by K1, Nef or both were examined for their proangiogenic effects in Matrigel plug assay in nude mice as described in the ‘Materials and Methods’ section. Representative photographs of angiogenesis in the nude mice are shown. (H) The hemoglobin level of the Matrigel plugs treated as in (G) was determined with O.D. value at 540 nm. Data represent mean ± SD, each group with five tumors (n = 5). Three independent experiments were performed and similar results were obtained.

Mentions: Based on the above findings in vitro, we examined the synergistic effect of Nef and K1 on angiogenesis in the CAM model. Compared to control cells (Mock + PBS), lentivirus-mediated K1 expression and addition of soluble Nef protein alone in HUVECs promoted angiogenesis by 1.61- and 1.44-fold in the CAM model (Figure 2A and B). K1 and Nef protein synergized with each other and increased the angiogenic index by 4.25-fold. Similar results were also observed with ectopically Nef-expressing EA.hy926 cells (Figure 2C).


HIV-1 Nef and KSHV oncogene K1 synergistically promote angiogenesis by inducing cellular miR-718 to regulate the PTEN/AKT/mTOR signaling pathway.

Xue M, Yao S, Hu M, Li W, Hao T, Zhou F, Zhu X, Lu H, Qin D, Yan Q, Zhu J, Gao SJ, Lu C - Nucleic Acids Res. (2014)

Nef promotes K1-induced angiogenesis and tumorigenesis in the CAM and nude mice models. (A) Nef promoted K1-induced angiogenesis in HUVECs. HUVECs transduced with K1, incubated with soluble Nef protein for 72 h or both were mixed with Matrigel and subsequently implanted onto the CAM. Representative photographs of angiogenesis on the CAM are shown. (B) Quantification of results in (A). The number of blood vessels was normalized to that of Matrigel alone. Data represent mean ± SD from three independent experiments (n = 3), each experiment containing 10 technical replicates. (C) Nef enhanced K1-induced angiogenesis in EA.hy926 cells. EA.hy926 cells transduced with K1, Nef or both were mixed with Matrigel and subsequently implanted onto the CAM. The number of blood vessels was normalized to that of Matrigel alone. Data represent mean ± SD from three independent experiments (n = 3), each experiment containing 10 technical replicates. (D) Hematoxylin and eosin staining analysis of histologic features (top; original magnification, ×100) and immunohistochemical staining analysis of the expression of SMA and VEGF (middle and bottom; original magnification, ×200) in tumor tissues from the CAMs induced by EA.hy926 cells transduced with K1 and Nef. Black arrows point to neovascularization and hemorrhagic foci. (E) Quantification of results in (D). (F) Western blot analysis of total PTEN and phosphorylation levels of PI3K, AKT and mTOR in tumor tissues from the CAMs treated as in (D). (G) Nef enhanced K1-induced angiogenesis in nude mice. EA.hy926 cells transduced by K1, Nef or both were examined for their proangiogenic effects in Matrigel plug assay in nude mice as described in the ‘Materials and Methods’ section. Representative photographs of angiogenesis in the nude mice are shown. (H) The hemoglobin level of the Matrigel plugs treated as in (G) was determined with O.D. value at 540 nm. Data represent mean ± SD, each group with five tumors (n = 5). Three independent experiments were performed and similar results were obtained.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 2: Nef promotes K1-induced angiogenesis and tumorigenesis in the CAM and nude mice models. (A) Nef promoted K1-induced angiogenesis in HUVECs. HUVECs transduced with K1, incubated with soluble Nef protein for 72 h or both were mixed with Matrigel and subsequently implanted onto the CAM. Representative photographs of angiogenesis on the CAM are shown. (B) Quantification of results in (A). The number of blood vessels was normalized to that of Matrigel alone. Data represent mean ± SD from three independent experiments (n = 3), each experiment containing 10 technical replicates. (C) Nef enhanced K1-induced angiogenesis in EA.hy926 cells. EA.hy926 cells transduced with K1, Nef or both were mixed with Matrigel and subsequently implanted onto the CAM. The number of blood vessels was normalized to that of Matrigel alone. Data represent mean ± SD from three independent experiments (n = 3), each experiment containing 10 technical replicates. (D) Hematoxylin and eosin staining analysis of histologic features (top; original magnification, ×100) and immunohistochemical staining analysis of the expression of SMA and VEGF (middle and bottom; original magnification, ×200) in tumor tissues from the CAMs induced by EA.hy926 cells transduced with K1 and Nef. Black arrows point to neovascularization and hemorrhagic foci. (E) Quantification of results in (D). (F) Western blot analysis of total PTEN and phosphorylation levels of PI3K, AKT and mTOR in tumor tissues from the CAMs treated as in (D). (G) Nef enhanced K1-induced angiogenesis in nude mice. EA.hy926 cells transduced by K1, Nef or both were examined for their proangiogenic effects in Matrigel plug assay in nude mice as described in the ‘Materials and Methods’ section. Representative photographs of angiogenesis in the nude mice are shown. (H) The hemoglobin level of the Matrigel plugs treated as in (G) was determined with O.D. value at 540 nm. Data represent mean ± SD, each group with five tumors (n = 5). Three independent experiments were performed and similar results were obtained.
Mentions: Based on the above findings in vitro, we examined the synergistic effect of Nef and K1 on angiogenesis in the CAM model. Compared to control cells (Mock + PBS), lentivirus-mediated K1 expression and addition of soluble Nef protein alone in HUVECs promoted angiogenesis by 1.61- and 1.44-fold in the CAM model (Figure 2A and B). K1 and Nef protein synergized with each other and increased the angiogenic index by 4.25-fold. Similar results were also observed with ectopically Nef-expressing EA.hy926 cells (Figure 2C).

Bottom Line: Kaposi's sarcoma (KS) is an AIDS-defining cancer with aberrant neovascularization caused by KS-associated herpesvirus (KSHV).Furthermore, Nef and K1 induced cellular miR-718, which inhibited PTEN expression by directly targeting a seed sequence in the 3' UTR of its mRNA.Our results demonstrate an essential role of miR-718/AKT/mTOR axis in AIDS-KS and thus may represent an attractive therapeutic target.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, P.R. China Key Laboratory of Pathogen Biology of Jiangsu Province, Nanjing Medical University, Nanjing, P.R. China Department of Microbiology, Nanjing Medical University, Nanjing 210029, P.R. China Department of Physiology, Xuzhou Medical College, Xuzhou, Jiangsu 221004, P.R. China.

Show MeSH
Related in: MedlinePlus