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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 MMP2 activity and decreases expression levels of metastasis-related genes. (A) CEP65 promotes MMP2 activity. Serum-free conditioned media from indicated cells were subjected to zymographic assay. Following removal of the SDS, the gelatin-containing gel was incubated in developing buffer and then visualized by Coomassie blue staining (upper panel). Relative activities of MMP2 and MMP9 were calculated (lower panel). (B) Effect of CEP65 on gene expression, as detected by reverse transcription polymerase chain reaction. **P<0.01 vs. AGS; ##P<0.01 vs. AGS-M. CEP65, cancer and embryo expression protein 65; MMP, matrix metalloproteinase; GAPDH, glyceraldehyde 3-phosphate; AGS-M, AGS mock-transfected cells.
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f3-ol-09-04-1772: CEP65 promotes MMP2 activity and decreases expression levels of metastasis-related genes. (A) CEP65 promotes MMP2 activity. Serum-free conditioned media from indicated cells were subjected to zymographic assay. Following removal of the SDS, the gelatin-containing gel was incubated in developing buffer and then visualized by Coomassie blue staining (upper panel). Relative activities of MMP2 and MMP9 were calculated (lower panel). (B) Effect of CEP65 on gene expression, as detected by reverse transcription polymerase chain reaction. **P<0.01 vs. AGS; ##P<0.01 vs. AGS-M. CEP65, cancer and embryo expression protein 65; MMP, matrix metalloproteinase; GAPDH, glyceraldehyde 3-phosphate; AGS-M, AGS mock-transfected cells.

Mentions: The results of the present study suggested that CEP65 has a proinvasive function in cancer cells, while the mechanism underlying this role remains unclear. Since the degradation of the ECM by MMPs is essential in metastasis (11), the activities of MMP2 and MMP9 in the culture supernatant of AGS cells were investigated by zymographic assay. As shown in Fig. 3A, the collagen-degrading activity of MMP2 was higher in the AGS-CEP65 cell supernatant compared with the activity in the AGS and AGS-M cells (P<0.01), however, CEP65 did not exhibit a stimulatory effect on the activity of MMP9. The mRNA expression levels of MMP2 and MMP9 were not affected by CEP65 (Fig. 3B). Therefore, the increased MMP2 activity in the conditioned medium of AGS-CEP65 cells did not result from the upregulation of MMP2 expression.


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 MMP2 activity and decreases expression levels of metastasis-related genes. (A) CEP65 promotes MMP2 activity. Serum-free conditioned media from indicated cells were subjected to zymographic assay. Following removal of the SDS, the gelatin-containing gel was incubated in developing buffer and then visualized by Coomassie blue staining (upper panel). Relative activities of MMP2 and MMP9 were calculated (lower panel). (B) Effect of CEP65 on gene expression, as detected by reverse transcription polymerase chain reaction. **P<0.01 vs. AGS; ##P<0.01 vs. AGS-M. CEP65, cancer and embryo expression protein 65; MMP, matrix metalloproteinase; GAPDH, glyceraldehyde 3-phosphate; AGS-M, AGS mock-transfected cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356324&req=5

f3-ol-09-04-1772: CEP65 promotes MMP2 activity and decreases expression levels of metastasis-related genes. (A) CEP65 promotes MMP2 activity. Serum-free conditioned media from indicated cells were subjected to zymographic assay. Following removal of the SDS, the gelatin-containing gel was incubated in developing buffer and then visualized by Coomassie blue staining (upper panel). Relative activities of MMP2 and MMP9 were calculated (lower panel). (B) Effect of CEP65 on gene expression, as detected by reverse transcription polymerase chain reaction. **P<0.01 vs. AGS; ##P<0.01 vs. AGS-M. CEP65, cancer and embryo expression protein 65; MMP, matrix metalloproteinase; GAPDH, glyceraldehyde 3-phosphate; AGS-M, AGS mock-transfected cells.
Mentions: The results of the present study suggested that CEP65 has a proinvasive function in cancer cells, while the mechanism underlying this role remains unclear. Since the degradation of the ECM by MMPs is essential in metastasis (11), the activities of MMP2 and MMP9 in the culture supernatant of AGS cells were investigated by zymographic assay. As shown in Fig. 3A, the collagen-degrading activity of MMP2 was higher in the AGS-CEP65 cell supernatant compared with the activity in the AGS and AGS-M cells (P<0.01), however, CEP65 did not exhibit a stimulatory effect on the activity of MMP9. The mRNA expression levels of MMP2 and MMP9 were not affected by CEP65 (Fig. 3B). Therefore, the increased MMP2 activity in the conditioned medium of AGS-CEP65 cells did not result from the upregulation of MMP2 expression.

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