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Mesenchymal stem cells promote colorectal cancer progression through AMPK/mTOR-mediated NF-κB activation.

Wu XB, Liu Y, Wang GH, Xu X, Cai Y, Wang HY, Li YQ, Meng HF, Dai F, Jin JD - Sci Rep (2016)

Bottom Line: This study was designed to identify the molecular mechanisms related to the tumor-promoting effect of MSCs in colorectal cancer.The tumorigenic effect of MSC-CM was attributed to altered expression of cell cycle regulatory proteins and inhibition of apoptosis.This study indicates that MSCs promote the progression of colorectal cancer via AMPK/mTOR-mediated NF-κB activation.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, the Third Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230061, P. R. China.

ABSTRACT
Mesenchymal stem cells (MSCs) exert a tumor-promoting effect in a variety of human cancers. This study was designed to identify the molecular mechanisms related to the tumor-promoting effect of MSCs in colorectal cancer. In vitro analysis of colorectal cancer cell lines cultured in MSC conditioned media (MSC-CM) showed that MSC-CM significantly promoted the progression of the cancer cells by enhancing cell proliferation, migration and colony formation. The tumorigenic effect of MSC-CM was attributed to altered expression of cell cycle regulatory proteins and inhibition of apoptosis. Furthermore, MSC-CM induced high level expression of a number of pluripotency factors in the cancer cells. ELISAs revealed MSC-CM contained higher levels of IL-6 and IL-8, which are associated with the progression of cancer. Moreover, MSC-CM downregulated AMPK mRNA and protein phosphorylation, but upregulated mTOR mRNA and protein phosphorylation. The NF-κB pathway was activated after addition of MSC-CM. An in vivo model in Balb/C mice confirmed the ability of MSC-CM to promote the invasion and proliferation of colorectal cancer cells. This study indicates that MSCs promote the progression of colorectal cancer via AMPK/mTOR-mediated NF-κB activation.

No MeSH data available.


Related in: MedlinePlus

Conditioned media from MSCs regulates the cell cycle and inhibits apoptosis in CRC cell lines.HCT116 and LOVO cells were cultured in MSC-CM or control media for 24 h. (A) Flow cytometric analysis of JC-1 expression. (B) Western blot analysis of P53, Bax and Bcl-2 expression. (C) Flow cytometric analysis of cell cycle distribution in HCT116 cells. (D) Quantification of P53, Bax and Bcl-2 expression from Western blots. (E) Q-PCR analysis of P53, P21 and P16 mRNA expression. *P < 0.05 and **P < 0.01 vs. control group (n > 3).
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f3: Conditioned media from MSCs regulates the cell cycle and inhibits apoptosis in CRC cell lines.HCT116 and LOVO cells were cultured in MSC-CM or control media for 24 h. (A) Flow cytometric analysis of JC-1 expression. (B) Western blot analysis of P53, Bax and Bcl-2 expression. (C) Flow cytometric analysis of cell cycle distribution in HCT116 cells. (D) Quantification of P53, Bax and Bcl-2 expression from Western blots. (E) Q-PCR analysis of P53, P21 and P16 mRNA expression. *P < 0.05 and **P < 0.01 vs. control group (n > 3).

Mentions: The mitochondrial membrane potential assay was used to determine whether MSCs can promote CRC by inhibiting apoptosis. JC-1 staining was analyzed using flow cytometry and showed that culture in MSC-CM significantly decreased the rate of apoptosis in HCT116 cells compared with culture in control media (Fig. 3A). Western blotting demonstrated that the presence of MSC-CM downregulated the expression of the apoptosis-related proteins Bax and P53 and upregulated the anti-apoptotic protein Bcl-2 (Fig. 3B,D)compared to cells cultured in control media; the full-length blots are displayed in Supplementary Figure S2. To determine if the pro-proliferative effects of MSC-CM on HCT116 cells were associated with promotion of the cell cycle, we stained the cells with propidium iodine (PI) to assess the cell cycle using flow cytometry. MSC-CM significantly altered the cell cycle distribution of diploid CRC cells, with a tendency towards an increased percentage of cells in the S phase of the cell cycle (Fig. 3C). To further analyze the effect of MSC-CM on cell cycle progression, we also assessed the protein expression levels of two factors that negatively regulate the cell cycle. Western blotting demonstrated that HCT116 cells cultured in MSC-CM expressed significantly lower mRNA levels of both P53,P16 and P21 compared to cells cultured in control media (Fig. 3E). These results suggest that MSCs promote proliferation in CRC via a mechanism associated with altered cell cycle progression and inhibition of cell apoptosis.


Mesenchymal stem cells promote colorectal cancer progression through AMPK/mTOR-mediated NF-κB activation.

Wu XB, Liu Y, Wang GH, Xu X, Cai Y, Wang HY, Li YQ, Meng HF, Dai F, Jin JD - Sci Rep (2016)

Conditioned media from MSCs regulates the cell cycle and inhibits apoptosis in CRC cell lines.HCT116 and LOVO cells were cultured in MSC-CM or control media for 24 h. (A) Flow cytometric analysis of JC-1 expression. (B) Western blot analysis of P53, Bax and Bcl-2 expression. (C) Flow cytometric analysis of cell cycle distribution in HCT116 cells. (D) Quantification of P53, Bax and Bcl-2 expression from Western blots. (E) Q-PCR analysis of P53, P21 and P16 mRNA expression. *P < 0.05 and **P < 0.01 vs. control group (n > 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4759824&req=5

f3: Conditioned media from MSCs regulates the cell cycle and inhibits apoptosis in CRC cell lines.HCT116 and LOVO cells were cultured in MSC-CM or control media for 24 h. (A) Flow cytometric analysis of JC-1 expression. (B) Western blot analysis of P53, Bax and Bcl-2 expression. (C) Flow cytometric analysis of cell cycle distribution in HCT116 cells. (D) Quantification of P53, Bax and Bcl-2 expression from Western blots. (E) Q-PCR analysis of P53, P21 and P16 mRNA expression. *P < 0.05 and **P < 0.01 vs. control group (n > 3).
Mentions: The mitochondrial membrane potential assay was used to determine whether MSCs can promote CRC by inhibiting apoptosis. JC-1 staining was analyzed using flow cytometry and showed that culture in MSC-CM significantly decreased the rate of apoptosis in HCT116 cells compared with culture in control media (Fig. 3A). Western blotting demonstrated that the presence of MSC-CM downregulated the expression of the apoptosis-related proteins Bax and P53 and upregulated the anti-apoptotic protein Bcl-2 (Fig. 3B,D)compared to cells cultured in control media; the full-length blots are displayed in Supplementary Figure S2. To determine if the pro-proliferative effects of MSC-CM on HCT116 cells were associated with promotion of the cell cycle, we stained the cells with propidium iodine (PI) to assess the cell cycle using flow cytometry. MSC-CM significantly altered the cell cycle distribution of diploid CRC cells, with a tendency towards an increased percentage of cells in the S phase of the cell cycle (Fig. 3C). To further analyze the effect of MSC-CM on cell cycle progression, we also assessed the protein expression levels of two factors that negatively regulate the cell cycle. Western blotting demonstrated that HCT116 cells cultured in MSC-CM expressed significantly lower mRNA levels of both P53,P16 and P21 compared to cells cultured in control media (Fig. 3E). These results suggest that MSCs promote proliferation in CRC via a mechanism associated with altered cell cycle progression and inhibition of cell apoptosis.

Bottom Line: This study was designed to identify the molecular mechanisms related to the tumor-promoting effect of MSCs in colorectal cancer.The tumorigenic effect of MSC-CM was attributed to altered expression of cell cycle regulatory proteins and inhibition of apoptosis.This study indicates that MSCs promote the progression of colorectal cancer via AMPK/mTOR-mediated NF-κB activation.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, the Third Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230061, P. R. China.

ABSTRACT
Mesenchymal stem cells (MSCs) exert a tumor-promoting effect in a variety of human cancers. This study was designed to identify the molecular mechanisms related to the tumor-promoting effect of MSCs in colorectal cancer. In vitro analysis of colorectal cancer cell lines cultured in MSC conditioned media (MSC-CM) showed that MSC-CM significantly promoted the progression of the cancer cells by enhancing cell proliferation, migration and colony formation. The tumorigenic effect of MSC-CM was attributed to altered expression of cell cycle regulatory proteins and inhibition of apoptosis. Furthermore, MSC-CM induced high level expression of a number of pluripotency factors in the cancer cells. ELISAs revealed MSC-CM contained higher levels of IL-6 and IL-8, which are associated with the progression of cancer. Moreover, MSC-CM downregulated AMPK mRNA and protein phosphorylation, but upregulated mTOR mRNA and protein phosphorylation. The NF-κB pathway was activated after addition of MSC-CM. An in vivo model in Balb/C mice confirmed the ability of MSC-CM to promote the invasion and proliferation of colorectal cancer cells. This study indicates that MSCs promote the progression of colorectal cancer via AMPK/mTOR-mediated NF-κB activation.

No MeSH data available.


Related in: MedlinePlus