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Cancer and embryo expression protein 65 promotes cancer cell growth and metastasis.

Jin G, Peng L, Zhang J, Qu L, Shou C - Oncol Lett (2015)

Bottom Line: By inoculating BICR-H1 cells on chick chorioallantoic membrane (CAM), it was found that CEP65 promotes cell growth on the CAM and increases cell metastasis to the lungs of the chicken.By utilizing a xenograft severe combined immunodeficiency mouse model, CEP65 was also found to accelerate BICR-H1 cell growth and metastasis to the lungs.Taken together, the results of the present study demonstrated the oncogenic function of CEP65 in promoting cancer cell growth and metastasis.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Biochemistry and Molecular Biology, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China.

ABSTRACT

Cancer and embryo expression protein 65 (CEP65) is a centrosomal protein that is expressed at relatively high levels in embryonic tissue and different cancerous tissues, but its role in tumorigenesis remains unknown. In the present study, CEP65 was stably expressed in AGS gastric cancer cells. CEP65 was found to promote cell growth in the MTT assay and to enhance cell migration and invasion in Transwell chamber assays. To validate results from the in vitro experiments, CEP65 was stably expressed in BICR-H1 breast cancer cells through adenovirus-mediated transduction. By inoculating BICR-H1 cells on chick chorioallantoic membrane (CAM), it was found that CEP65 promotes cell growth on the CAM and increases cell metastasis to the lungs of the chicken. By utilizing a xenograft severe combined immunodeficiency mouse model, CEP65 was also found to accelerate BICR-H1 cell growth and metastasis to the lungs. Furthermore, it was shown that CEP65 increases matrix metalloproteinase (MMP)2 activity in zymographic assays, however, microarray screening and reverse transcription polymerase chain reaction validation revealed that CEP65 had no effect on the expression levels of MMP2 or MMP9, but decreased the expression levels of metastasis-associated genes, TIMP2, RAP and VTN. Taken together, the results of the present study demonstrated the oncogenic function of CEP65 in promoting cancer cell growth and metastasis.

No MeSH data available.


Related in: MedlinePlus

CEP65 promotes BICR-H1 cells growth and metastasis in vivo. (A) Expression of CEP65 in adenovirus-infected BICR-H1 cells, as detected by western blot analysis. (B) CEP65 promotes tumor formation on chick chorioallantoic membranes: Left image, macroscopic image of tumors; right image, graph summarizing the weight of the tumors. (C) CEP65 promotes BICR-H1 metastasis to the lungs of the chicken: Left, metastatic foci in the chicken lungs were detected using fluorescence microscopy; right, graph summarizing the numbers of metastatic foci. (D) CEP65 promotes tumor formation in SCID mice: Left, macroscopic image of tumors; graph summarizing the weight of the tumors. (E) CEP65 promotes BICR-H1 metastasis to the lungs of SCID mice, as detected by hematoxylin and eosin staining. *P<0.05 vs. pAd. CEP65, cancer and embryo expression protein 65; SCID, severe combined immunodeficiency.
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f2-ol-09-04-1772: CEP65 promotes BICR-H1 cells growth and metastasis in vivo. (A) Expression of CEP65 in adenovirus-infected BICR-H1 cells, as detected by western blot analysis. (B) CEP65 promotes tumor formation on chick chorioallantoic membranes: Left image, macroscopic image of tumors; right image, graph summarizing the weight of the tumors. (C) CEP65 promotes BICR-H1 metastasis to the lungs of the chicken: Left, metastatic foci in the chicken lungs were detected using fluorescence microscopy; right, graph summarizing the numbers of metastatic foci. (D) CEP65 promotes tumor formation in SCID mice: Left, macroscopic image of tumors; graph summarizing the weight of the tumors. (E) CEP65 promotes BICR-H1 metastasis to the lungs of SCID mice, as detected by hematoxylin and eosin staining. *P<0.05 vs. pAd. CEP65, cancer and embryo expression protein 65; SCID, severe combined immunodeficiency.

Mentions: To validate the results from the in vitro assays, the effects of CEP65 on cancer cell growth and metastasis were further investigated in vivo. Due to the low capacity of AGS cells to form metastatic foci in animals, BICR-H1 cells were used in the present study; these are breast cancer cells with defined metastatic ability that have been used in a previous study (9). CEP65 was barely detectable in the BICR-H1 cells, using western blot analysis (Fig. 2A). Adenovirus-mediated transduction of antisense CEP65 [pAd-CEP65(−)] resulted in the complete elimination of CEP65 expression (Fig. 2A), while transduction of sense CEP65 [pAd-CEP65(+)] was found to strongly increase the CEP65 protein levels (Fig. 2A). Following inoculation of the plasmids on the chick CAMs for seven days, tumors were formed (Fig. 2B). The tumors were weighed, revealing that pAd-CEP65(+) increased tumor growth (P<0.05), whereas pAd-CEP65(−) decreased tumor growth (P<0.05) (Fig. 2B). Using fluorescence microscopy, the metastasis of the BICR-H1 cells to the chicken embryo lungs was increased by 5.3-fold in the cells infected with pAd-CEP65(+) as compared with the cells infected with pAd (P<0.05). Metastasis was found to be reduced by 76.2% in the cells infected with pAd-CEP65(−) as compared with the cells infected with pAd (P<0.05; Fig. 2C). Furthermore, infected BICR-H1 cells were transfected into the mammary fat pads of SCID mice and the CEP65 expression levels were found to be positively correlated with cell growth (Fig. 2D) and metastatic foci formation in the lungs (Fig. 2E). These results indicated that CEP65 promoted the growth and metastasis of the BICR-H1 cells in vivo.


Cancer and embryo expression protein 65 promotes cancer cell growth and metastasis.

Jin G, Peng L, Zhang J, Qu L, Shou C - Oncol Lett (2015)

CEP65 promotes BICR-H1 cells growth and metastasis in vivo. (A) Expression of CEP65 in adenovirus-infected BICR-H1 cells, as detected by western blot analysis. (B) CEP65 promotes tumor formation on chick chorioallantoic membranes: Left image, macroscopic image of tumors; right image, graph summarizing the weight of the tumors. (C) CEP65 promotes BICR-H1 metastasis to the lungs of the chicken: Left, metastatic foci in the chicken lungs were detected using fluorescence microscopy; right, graph summarizing the numbers of metastatic foci. (D) CEP65 promotes tumor formation in SCID mice: Left, macroscopic image of tumors; graph summarizing the weight of the tumors. (E) CEP65 promotes BICR-H1 metastasis to the lungs of SCID mice, as detected by hematoxylin and eosin staining. *P<0.05 vs. pAd. CEP65, cancer and embryo expression protein 65; SCID, severe combined immunodeficiency.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4356324&req=5

f2-ol-09-04-1772: CEP65 promotes BICR-H1 cells growth and metastasis in vivo. (A) Expression of CEP65 in adenovirus-infected BICR-H1 cells, as detected by western blot analysis. (B) CEP65 promotes tumor formation on chick chorioallantoic membranes: Left image, macroscopic image of tumors; right image, graph summarizing the weight of the tumors. (C) CEP65 promotes BICR-H1 metastasis to the lungs of the chicken: Left, metastatic foci in the chicken lungs were detected using fluorescence microscopy; right, graph summarizing the numbers of metastatic foci. (D) CEP65 promotes tumor formation in SCID mice: Left, macroscopic image of tumors; graph summarizing the weight of the tumors. (E) CEP65 promotes BICR-H1 metastasis to the lungs of SCID mice, as detected by hematoxylin and eosin staining. *P<0.05 vs. pAd. CEP65, cancer and embryo expression protein 65; SCID, severe combined immunodeficiency.
Mentions: To validate the results from the in vitro assays, the effects of CEP65 on cancer cell growth and metastasis were further investigated in vivo. Due to the low capacity of AGS cells to form metastatic foci in animals, BICR-H1 cells were used in the present study; these are breast cancer cells with defined metastatic ability that have been used in a previous study (9). CEP65 was barely detectable in the BICR-H1 cells, using western blot analysis (Fig. 2A). Adenovirus-mediated transduction of antisense CEP65 [pAd-CEP65(−)] resulted in the complete elimination of CEP65 expression (Fig. 2A), while transduction of sense CEP65 [pAd-CEP65(+)] was found to strongly increase the CEP65 protein levels (Fig. 2A). Following inoculation of the plasmids on the chick CAMs for seven days, tumors were formed (Fig. 2B). The tumors were weighed, revealing that pAd-CEP65(+) increased tumor growth (P<0.05), whereas pAd-CEP65(−) decreased tumor growth (P<0.05) (Fig. 2B). Using fluorescence microscopy, the metastasis of the BICR-H1 cells to the chicken embryo lungs was increased by 5.3-fold in the cells infected with pAd-CEP65(+) as compared with the cells infected with pAd (P<0.05). Metastasis was found to be reduced by 76.2% in the cells infected with pAd-CEP65(−) as compared with the cells infected with pAd (P<0.05; Fig. 2C). Furthermore, infected BICR-H1 cells were transfected into the mammary fat pads of SCID mice and the CEP65 expression levels were found to be positively correlated with cell growth (Fig. 2D) and metastatic foci formation in the lungs (Fig. 2E). These results indicated that CEP65 promoted the growth and metastasis of the BICR-H1 cells in vivo.

Bottom Line: By inoculating BICR-H1 cells on chick chorioallantoic membrane (CAM), it was found that CEP65 promotes cell growth on the CAM and increases cell metastasis to the lungs of the chicken.By utilizing a xenograft severe combined immunodeficiency mouse model, CEP65 was also found to accelerate BICR-H1 cell growth and metastasis to the lungs.Taken together, the results of the present study demonstrated the oncogenic function of CEP65 in promoting cancer cell growth and metastasis.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Biochemistry and Molecular Biology, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China.

ABSTRACT

Cancer and embryo expression protein 65 (CEP65) is a centrosomal protein that is expressed at relatively high levels in embryonic tissue and different cancerous tissues, but its role in tumorigenesis remains unknown. In the present study, CEP65 was stably expressed in AGS gastric cancer cells. CEP65 was found to promote cell growth in the MTT assay and to enhance cell migration and invasion in Transwell chamber assays. To validate results from the in vitro experiments, CEP65 was stably expressed in BICR-H1 breast cancer cells through adenovirus-mediated transduction. By inoculating BICR-H1 cells on chick chorioallantoic membrane (CAM), it was found that CEP65 promotes cell growth on the CAM and increases cell metastasis to the lungs of the chicken. By utilizing a xenograft severe combined immunodeficiency mouse model, CEP65 was also found to accelerate BICR-H1 cell growth and metastasis to the lungs. Furthermore, it was shown that CEP65 increases matrix metalloproteinase (MMP)2 activity in zymographic assays, however, microarray screening and reverse transcription polymerase chain reaction validation revealed that CEP65 had no effect on the expression levels of MMP2 or MMP9, but decreased the expression levels of metastasis-associated genes, TIMP2, RAP and VTN. Taken together, the results of the present study demonstrated the oncogenic function of CEP65 in promoting cancer cell growth and metastasis.

No MeSH data available.


Related in: MedlinePlus