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Modulator of apoptosis 1 (MOAP-1) is a tumor suppressor protein linked to the RASSF1A protein.

Law J, Salla M, Zare A, Wong Y, Luong L, Volodko N, Svystun O, Flood K, Lim J, Sung M, Dyck JR, Tan CT, Su YC, Yu VC, Mackey J, Baksh S - J. Biol. Chem. (2015)

Bottom Line: It is an integral partner to the tumor suppressor protein, Ras association domain family 1A (RASSF1A), and functions to activate the Bcl-2 family pro-apoptotic protein Bax.Although RASSF1A is now considered a bona fide tumor suppressor protein, the role of MOAP-1 as a tumor suppressor protein has yet to be determined.Overexpression of MOAP-1 in several cancer cell lines resulted in reduced tumorigenesis and up-regulation of genes involved in cancer regulatory pathways that include apoptosis (p53, Fas, and MST1), DNA damage control (poly(ADP)-ribose polymerase and ataxia telangiectasia mutated), those within the cell metabolism (IR-α, IR-β, and AMP-activated protein kinase), and a stabilizing effect on microtubules.

View Article: PubMed Central - PubMed

Affiliation: From the Departments of Biochemistry and.

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MOAP-1 can inhibit tumor formation in vivo. Xenograft assays were carried out in DAOY medulloblastoma and SKVO3 ovarian cancer cells (A and B) and in H1299 lung cancer cells (C and D). Stable cells were utilized for H1299 and SKOV3 cells, although DAOY cells contained transiently transfected expression vectors. For SKOV3, the anti-GFP antibody recognized both CFP-MOAP-1 and GFP-1A. For all, cells were harvested 48 h post-transient transfection, resuspended in Matrigel, and subcutaneously injected into male athymic mice. Immunoblots (IB) beside the graphs represent stable or transient expression of the indicated proteins. Athymic nude mice (Taconic Laboratories number NCRNU-M, CrTac:NCr-FoxN1Nu) were utilized for this assay as described previously (11). A, for DAOY cells, the p value between vector and HA-RASSF1A is 0.0003; the p value between vector and Myc-MOAP-1 or vector and Myc-MOAP-1 plus HA-RASSF1A is 0.0023 and 0.0065 respectively; n = 6–10 for all. A, right side, expressions of the indicated constructs are shown. For SKOV3, the p value between vector and HA-RASSF1A and vector and Myc-MOAP-1 was 0.009 and 0.004, respectively; n = 6 for all. C, p value between tumors formed in vector versus MOAP-1 cells was 0.0075, vector and RASSF1A was 0.0214, and vector and RASSF1A/MOAP-1 was 0.0049 (n = 8–10 for all). p value between MOAP-1 wild type and δBH3 or BH3 mutant is <0.027 and <0.0327, respectively.
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Figure 7: MOAP-1 can inhibit tumor formation in vivo. Xenograft assays were carried out in DAOY medulloblastoma and SKVO3 ovarian cancer cells (A and B) and in H1299 lung cancer cells (C and D). Stable cells were utilized for H1299 and SKOV3 cells, although DAOY cells contained transiently transfected expression vectors. For SKOV3, the anti-GFP antibody recognized both CFP-MOAP-1 and GFP-1A. For all, cells were harvested 48 h post-transient transfection, resuspended in Matrigel, and subcutaneously injected into male athymic mice. Immunoblots (IB) beside the graphs represent stable or transient expression of the indicated proteins. Athymic nude mice (Taconic Laboratories number NCRNU-M, CrTac:NCr-FoxN1Nu) were utilized for this assay as described previously (11). A, for DAOY cells, the p value between vector and HA-RASSF1A is 0.0003; the p value between vector and Myc-MOAP-1 or vector and Myc-MOAP-1 plus HA-RASSF1A is 0.0023 and 0.0065 respectively; n = 6–10 for all. A, right side, expressions of the indicated constructs are shown. For SKOV3, the p value between vector and HA-RASSF1A and vector and Myc-MOAP-1 was 0.009 and 0.004, respectively; n = 6 for all. C, p value between tumors formed in vector versus MOAP-1 cells was 0.0075, vector and RASSF1A was 0.0214, and vector and RASSF1A/MOAP-1 was 0.0049 (n = 8–10 for all). p value between MOAP-1 wild type and δBH3 or BH3 mutant is <0.027 and <0.0327, respectively.

Mentions: MOAP-1 can inhibit cell proliferation in vitro.A and B, MOAP-1 suppresses H1299 cell growth in culture. H1299 cells stably expressing vector or Myc-MOAP-1 (A) or MOAP-1 shRNA (B) were seeded at 4000 cells per well in a 96-well plate, and an MTT assay was carried out as outlined under “Experimental Procedures.” Inset represents immunoblot to confirm the expression of Myc-MOAP-1 with ERK1/2 as a loading control. A, p value = 0.0006, n = 16; B, p value = 0.0022, n = 8. B, right side, immunoblot to confirm shRNA knockdown of endogenous MOAP-1. IB, immunoblot. C, colony formation was carried out in H1299 stable cells expressing the indicated constructs. p value = 0.0361 and n = 4 −5 for both. H1299 cells were seeded at 600 cells per plate and allowed to grow for 11 days prior to staining with crystal violet. Number of cell colonies were counted by visual examination. The expression of Myc-MOAP-1 can be detected in these cells for >30 days after stables are formed (see Fig. 7C for stable expression of MOAP-1 in H1299).


Modulator of apoptosis 1 (MOAP-1) is a tumor suppressor protein linked to the RASSF1A protein.

Law J, Salla M, Zare A, Wong Y, Luong L, Volodko N, Svystun O, Flood K, Lim J, Sung M, Dyck JR, Tan CT, Su YC, Yu VC, Mackey J, Baksh S - J. Biol. Chem. (2015)

MOAP-1 can inhibit tumor formation in vivo. Xenograft assays were carried out in DAOY medulloblastoma and SKVO3 ovarian cancer cells (A and B) and in H1299 lung cancer cells (C and D). Stable cells were utilized for H1299 and SKOV3 cells, although DAOY cells contained transiently transfected expression vectors. For SKOV3, the anti-GFP antibody recognized both CFP-MOAP-1 and GFP-1A. For all, cells were harvested 48 h post-transient transfection, resuspended in Matrigel, and subcutaneously injected into male athymic mice. Immunoblots (IB) beside the graphs represent stable or transient expression of the indicated proteins. Athymic nude mice (Taconic Laboratories number NCRNU-M, CrTac:NCr-FoxN1Nu) were utilized for this assay as described previously (11). A, for DAOY cells, the p value between vector and HA-RASSF1A is 0.0003; the p value between vector and Myc-MOAP-1 or vector and Myc-MOAP-1 plus HA-RASSF1A is 0.0023 and 0.0065 respectively; n = 6–10 for all. A, right side, expressions of the indicated constructs are shown. For SKOV3, the p value between vector and HA-RASSF1A and vector and Myc-MOAP-1 was 0.009 and 0.004, respectively; n = 6 for all. C, p value between tumors formed in vector versus MOAP-1 cells was 0.0075, vector and RASSF1A was 0.0214, and vector and RASSF1A/MOAP-1 was 0.0049 (n = 8–10 for all). p value between MOAP-1 wild type and δBH3 or BH3 mutant is <0.027 and <0.0327, respectively.
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Figure 7: MOAP-1 can inhibit tumor formation in vivo. Xenograft assays were carried out in DAOY medulloblastoma and SKVO3 ovarian cancer cells (A and B) and in H1299 lung cancer cells (C and D). Stable cells were utilized for H1299 and SKOV3 cells, although DAOY cells contained transiently transfected expression vectors. For SKOV3, the anti-GFP antibody recognized both CFP-MOAP-1 and GFP-1A. For all, cells were harvested 48 h post-transient transfection, resuspended in Matrigel, and subcutaneously injected into male athymic mice. Immunoblots (IB) beside the graphs represent stable or transient expression of the indicated proteins. Athymic nude mice (Taconic Laboratories number NCRNU-M, CrTac:NCr-FoxN1Nu) were utilized for this assay as described previously (11). A, for DAOY cells, the p value between vector and HA-RASSF1A is 0.0003; the p value between vector and Myc-MOAP-1 or vector and Myc-MOAP-1 plus HA-RASSF1A is 0.0023 and 0.0065 respectively; n = 6–10 for all. A, right side, expressions of the indicated constructs are shown. For SKOV3, the p value between vector and HA-RASSF1A and vector and Myc-MOAP-1 was 0.009 and 0.004, respectively; n = 6 for all. C, p value between tumors formed in vector versus MOAP-1 cells was 0.0075, vector and RASSF1A was 0.0214, and vector and RASSF1A/MOAP-1 was 0.0049 (n = 8–10 for all). p value between MOAP-1 wild type and δBH3 or BH3 mutant is <0.027 and <0.0327, respectively.
Mentions: MOAP-1 can inhibit cell proliferation in vitro.A and B, MOAP-1 suppresses H1299 cell growth in culture. H1299 cells stably expressing vector or Myc-MOAP-1 (A) or MOAP-1 shRNA (B) were seeded at 4000 cells per well in a 96-well plate, and an MTT assay was carried out as outlined under “Experimental Procedures.” Inset represents immunoblot to confirm the expression of Myc-MOAP-1 with ERK1/2 as a loading control. A, p value = 0.0006, n = 16; B, p value = 0.0022, n = 8. B, right side, immunoblot to confirm shRNA knockdown of endogenous MOAP-1. IB, immunoblot. C, colony formation was carried out in H1299 stable cells expressing the indicated constructs. p value = 0.0361 and n = 4 −5 for both. H1299 cells were seeded at 600 cells per plate and allowed to grow for 11 days prior to staining with crystal violet. Number of cell colonies were counted by visual examination. The expression of Myc-MOAP-1 can be detected in these cells for >30 days after stables are formed (see Fig. 7C for stable expression of MOAP-1 in H1299).

Bottom Line: It is an integral partner to the tumor suppressor protein, Ras association domain family 1A (RASSF1A), and functions to activate the Bcl-2 family pro-apoptotic protein Bax.Although RASSF1A is now considered a bona fide tumor suppressor protein, the role of MOAP-1 as a tumor suppressor protein has yet to be determined.Overexpression of MOAP-1 in several cancer cell lines resulted in reduced tumorigenesis and up-regulation of genes involved in cancer regulatory pathways that include apoptosis (p53, Fas, and MST1), DNA damage control (poly(ADP)-ribose polymerase and ataxia telangiectasia mutated), those within the cell metabolism (IR-α, IR-β, and AMP-activated protein kinase), and a stabilizing effect on microtubules.

View Article: PubMed Central - PubMed

Affiliation: From the Departments of Biochemistry and.

Show MeSH
Related in: MedlinePlus