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CMTM3 inhibits cell migration and invasion and correlates with favorable prognosis in gastric cancer.

Su Y, Lin Y, Zhang L, Liu B, Yuan W, Mo X, Wang X, Li H, Xing X, Cheng X, Dong B, Hu Y, Du H, Zhu Y, Ding N, Li J, Liu W, Ma Y, Qiu X, Ji J, Han W - Cancer Sci. (2013)

Bottom Line: Restoration of CMTM3 significantly affected migration and invasion of AGS and SGC-7901 cells (P < 0.001).In vivo experiments showed that peritoneal disseminated metastases were significantly suppressed by CMTM3 (P < 0.001).We further showed that the expression of MMP2 and the phosphorylation of Erk1/2 were decreased when CMTM3 was restored.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, School of Basic Medical Sciences, Peking University Health Science Center, Peking University Center for Human Disease Genomics, Beijing, China; Key Laboratory of Medical Immunology, Ministry of Health, Beijing, China.

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(a) Real-time PCR was carried out to detect MMP2 expression in AGS and SGC-7901 gastric cancer cells. Average percentage of control (Mock) with SEM is shown from three independent experiments. (b) MMP2 in AGS cell supernatants was observed by gelatin zymography. Average relative gray density with SEM is shown from three independent experiments. (c) MMP2, phosphorylated Erk1/2 (P-Erk1/2) and total Erk1/2 (T-Erk1/2) in two cell lines were observed by Western blotting. The experiment was repeated at least three times and one representative result is shown. *P < 0.05; **P < 0.01.
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fig03: (a) Real-time PCR was carried out to detect MMP2 expression in AGS and SGC-7901 gastric cancer cells. Average percentage of control (Mock) with SEM is shown from three independent experiments. (b) MMP2 in AGS cell supernatants was observed by gelatin zymography. Average relative gray density with SEM is shown from three independent experiments. (c) MMP2, phosphorylated Erk1/2 (P-Erk1/2) and total Erk1/2 (T-Erk1/2) in two cell lines were observed by Western blotting. The experiment was repeated at least three times and one representative result is shown. *P < 0.05; **P < 0.01.

Mentions: To investigate the mechanism of CMTM3 as a tumor suppressor in gastric cancer cells, we collected AGS cells infected with Ad-Mock and Ad-CMTM3 and carried out the Tumor Metastasis RT Profiler PCR Array (SA Biosciences, Shanghai, China) which includes 84 genes known to be involved in cell migration and invasion. The PCR array results indicated that CMTM3 restoration may induce downregulation of several MMPs, especially MMP2 and MMP9. Then we measured the expression level of MMP2 and MMP9 in AGS cells and SGC-7901 cells by real-time PCR, and found that MMP2 expression was downregulated by CMTM3 significantly (Fig.3a), whereas the level of MMP9 was too low to be detected. Then gelatin zymography showed that CMTM3 inhibited the MMP2 level in cell supernatants and the gray density was significantly different (Fig.3b). To further confirm this result, we used Western blotting and found that the protein level of MMP2 from cell lysates was also reduced. These observations suggest that CMTM3 inhibits the expression and activity of MMP2. We also detected phosphorylated Erk1/2 (p-Erk1/2), a main signaling pathway regulating MMP2 expression. As shown in Figure3(c), p-Erk was remarkably reduced when CMTM3 was overexpressed, which indicated that CMTM3 might regulate MMP2 expression through the Erk1/2 signaling pathway.


CMTM3 inhibits cell migration and invasion and correlates with favorable prognosis in gastric cancer.

Su Y, Lin Y, Zhang L, Liu B, Yuan W, Mo X, Wang X, Li H, Xing X, Cheng X, Dong B, Hu Y, Du H, Zhu Y, Ding N, Li J, Liu W, Ma Y, Qiu X, Ji J, Han W - Cancer Sci. (2013)

(a) Real-time PCR was carried out to detect MMP2 expression in AGS and SGC-7901 gastric cancer cells. Average percentage of control (Mock) with SEM is shown from three independent experiments. (b) MMP2 in AGS cell supernatants was observed by gelatin zymography. Average relative gray density with SEM is shown from three independent experiments. (c) MMP2, phosphorylated Erk1/2 (P-Erk1/2) and total Erk1/2 (T-Erk1/2) in two cell lines were observed by Western blotting. The experiment was repeated at least three times and one representative result is shown. *P < 0.05; **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317871&req=5

fig03: (a) Real-time PCR was carried out to detect MMP2 expression in AGS and SGC-7901 gastric cancer cells. Average percentage of control (Mock) with SEM is shown from three independent experiments. (b) MMP2 in AGS cell supernatants was observed by gelatin zymography. Average relative gray density with SEM is shown from three independent experiments. (c) MMP2, phosphorylated Erk1/2 (P-Erk1/2) and total Erk1/2 (T-Erk1/2) in two cell lines were observed by Western blotting. The experiment was repeated at least three times and one representative result is shown. *P < 0.05; **P < 0.01.
Mentions: To investigate the mechanism of CMTM3 as a tumor suppressor in gastric cancer cells, we collected AGS cells infected with Ad-Mock and Ad-CMTM3 and carried out the Tumor Metastasis RT Profiler PCR Array (SA Biosciences, Shanghai, China) which includes 84 genes known to be involved in cell migration and invasion. The PCR array results indicated that CMTM3 restoration may induce downregulation of several MMPs, especially MMP2 and MMP9. Then we measured the expression level of MMP2 and MMP9 in AGS cells and SGC-7901 cells by real-time PCR, and found that MMP2 expression was downregulated by CMTM3 significantly (Fig.3a), whereas the level of MMP9 was too low to be detected. Then gelatin zymography showed that CMTM3 inhibited the MMP2 level in cell supernatants and the gray density was significantly different (Fig.3b). To further confirm this result, we used Western blotting and found that the protein level of MMP2 from cell lysates was also reduced. These observations suggest that CMTM3 inhibits the expression and activity of MMP2. We also detected phosphorylated Erk1/2 (p-Erk1/2), a main signaling pathway regulating MMP2 expression. As shown in Figure3(c), p-Erk was remarkably reduced when CMTM3 was overexpressed, which indicated that CMTM3 might regulate MMP2 expression through the Erk1/2 signaling pathway.

Bottom Line: Restoration of CMTM3 significantly affected migration and invasion of AGS and SGC-7901 cells (P < 0.001).In vivo experiments showed that peritoneal disseminated metastases were significantly suppressed by CMTM3 (P < 0.001).We further showed that the expression of MMP2 and the phosphorylation of Erk1/2 were decreased when CMTM3 was restored.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, School of Basic Medical Sciences, Peking University Health Science Center, Peking University Center for Human Disease Genomics, Beijing, China; Key Laboratory of Medical Immunology, Ministry of Health, Beijing, China.

Show MeSH
Related in: MedlinePlus