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The suppressing effects of BTG3 expression on aggressive behaviors and phenotypes of colorectal cancer: An in vitro and vivo study

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ABSTRACT

Here, we found that down-regulated expression of BTG3 might be positively correlated with colorectal carcinogenesis and its overexpression suppressed proliferation, glycolysis, mitochondrial respiration, cell cycle progression, migration, and invasion, and induced apoptosis, senescence and differentiation in SW480 and SW620 cells. After treated with cisplatin, MG132, paclitaxel and SAHA, BTG3 transfectants exhibited lower viability and higher apoptosis than the control in both time- and dose-dependent manners. BTG3 overexpression up- regulated the protein expression of Cyclin E, p16, p27, NF-κB, p38α/β, XIAP, Bcl-2, ATG14 and p53, but down-regulated the mRNA expression of MRP1, BCRP, and mTOR in SW480 and SW620 cells. BTG3 overexpression inhibited tumor growth of SW620 cells by suppressing proliferation and inducing apoptosis. It was suggested that down-regulated BTG3 expression might be considered as a marker for colorectal carcinogenesis. BTG3 overexpression might reverse the aggressive phenotypes and be employed as a potential target for gene therapy of colorectal cancer.

No MeSH data available.


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The effects of BTG3 overexpression on the phenotypes and their relevant molecules of colorectal cancer cellsAfter the transfection of pcDNA3.1-BTG3, its expression became strong in SW480 and SW620 cells by RT-PCR and Western blot A. The transfectants showed a low growth B. and G1 or S arrest C. in comparison with the control and mock. There was an apoptosis-induced effect of BTG3 overexpression in both transfectants, evidenced by Annexin V assay D. BTG3-overexpressing cells had a weaker ability to migrate and invade by wound healing E. and transwell assays F. BTG3 transfectants showed a higher senescence, a better differentiation, a lower level of glycolysis or mitochondrial function than the control and mock, evidenced by β-galactosidase staining G., ALP activity H., and metabolism assay I. respectively. The expression of phenotype- related molecules was screened by Western blot J. There was a high activity of NF-κB promoters in BTG3 transfectants, compared with the control and mock K. Results are representative of 3 different experiments, and data are expressed as mean ± standard deviation. Note: NC, negative control; * p < 0.05, compared with BTG3 transfectants.
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Figure 2: The effects of BTG3 overexpression on the phenotypes and their relevant molecules of colorectal cancer cellsAfter the transfection of pcDNA3.1-BTG3, its expression became strong in SW480 and SW620 cells by RT-PCR and Western blot A. The transfectants showed a low growth B. and G1 or S arrest C. in comparison with the control and mock. There was an apoptosis-induced effect of BTG3 overexpression in both transfectants, evidenced by Annexin V assay D. BTG3-overexpressing cells had a weaker ability to migrate and invade by wound healing E. and transwell assays F. BTG3 transfectants showed a higher senescence, a better differentiation, a lower level of glycolysis or mitochondrial function than the control and mock, evidenced by β-galactosidase staining G., ALP activity H., and metabolism assay I. respectively. The expression of phenotype- related molecules was screened by Western blot J. There was a high activity of NF-κB promoters in BTG3 transfectants, compared with the control and mock K. Results are representative of 3 different experiments, and data are expressed as mean ± standard deviation. Note: NC, negative control; * p < 0.05, compared with BTG3 transfectants.

Mentions: Here, we successfully transfected BTG3-expressing plasmid into SW480 and SW620 cells, evidenced by real-time RT-PCR and Western blot (Figure 2A). Both the transfectants showed a lower growth, evidenced by CCK-8 (Figure 2B, p<0.05) and a higher apoptosis by Annexin V-FITC staining in BTG3 transfectants than the control and mock (Figure 2D, p<0.05). There appeared G1 arrest in BTG 3-overxpressing SW480 cells in comparison to the control and mock by PI staining, while S arrest in SW620 cells (Figure 2C). BTG3 overexpression inhibited the migration and invasion of CRC cells according to wound healing (Figure 2E, p<0.05) and transwell chamber assays (Figure 2F, p<0.05), but induced senescence and differentiation, evidenced by β-galactosidase staining (Figure 2G) and alkaline phosphatase (ALP) activity (Figure 2H, p<0.05) respectively. BTG3-overexpressing CRC cells displayed a lower glycolysis and mitochondrial respiration than the mock and control according to oxygen consumption and extracellular acidification rates (Figure 2I, p<0.05).


The suppressing effects of BTG3 expression on aggressive behaviors and phenotypes of colorectal cancer: An in vitro and vivo study
The effects of BTG3 overexpression on the phenotypes and their relevant molecules of colorectal cancer cellsAfter the transfection of pcDNA3.1-BTG3, its expression became strong in SW480 and SW620 cells by RT-PCR and Western blot A. The transfectants showed a low growth B. and G1 or S arrest C. in comparison with the control and mock. There was an apoptosis-induced effect of BTG3 overexpression in both transfectants, evidenced by Annexin V assay D. BTG3-overexpressing cells had a weaker ability to migrate and invade by wound healing E. and transwell assays F. BTG3 transfectants showed a higher senescence, a better differentiation, a lower level of glycolysis or mitochondrial function than the control and mock, evidenced by β-galactosidase staining G., ALP activity H., and metabolism assay I. respectively. The expression of phenotype- related molecules was screened by Western blot J. There was a high activity of NF-κB promoters in BTG3 transfectants, compared with the control and mock K. Results are representative of 3 different experiments, and data are expressed as mean ± standard deviation. Note: NC, negative control; * p < 0.05, compared with BTG3 transfectants.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 2: The effects of BTG3 overexpression on the phenotypes and their relevant molecules of colorectal cancer cellsAfter the transfection of pcDNA3.1-BTG3, its expression became strong in SW480 and SW620 cells by RT-PCR and Western blot A. The transfectants showed a low growth B. and G1 or S arrest C. in comparison with the control and mock. There was an apoptosis-induced effect of BTG3 overexpression in both transfectants, evidenced by Annexin V assay D. BTG3-overexpressing cells had a weaker ability to migrate and invade by wound healing E. and transwell assays F. BTG3 transfectants showed a higher senescence, a better differentiation, a lower level of glycolysis or mitochondrial function than the control and mock, evidenced by β-galactosidase staining G., ALP activity H., and metabolism assay I. respectively. The expression of phenotype- related molecules was screened by Western blot J. There was a high activity of NF-κB promoters in BTG3 transfectants, compared with the control and mock K. Results are representative of 3 different experiments, and data are expressed as mean ± standard deviation. Note: NC, negative control; * p < 0.05, compared with BTG3 transfectants.
Mentions: Here, we successfully transfected BTG3-expressing plasmid into SW480 and SW620 cells, evidenced by real-time RT-PCR and Western blot (Figure 2A). Both the transfectants showed a lower growth, evidenced by CCK-8 (Figure 2B, p<0.05) and a higher apoptosis by Annexin V-FITC staining in BTG3 transfectants than the control and mock (Figure 2D, p<0.05). There appeared G1 arrest in BTG 3-overxpressing SW480 cells in comparison to the control and mock by PI staining, while S arrest in SW620 cells (Figure 2C). BTG3 overexpression inhibited the migration and invasion of CRC cells according to wound healing (Figure 2E, p<0.05) and transwell chamber assays (Figure 2F, p<0.05), but induced senescence and differentiation, evidenced by β-galactosidase staining (Figure 2G) and alkaline phosphatase (ALP) activity (Figure 2H, p<0.05) respectively. BTG3-overexpressing CRC cells displayed a lower glycolysis and mitochondrial respiration than the mock and control according to oxygen consumption and extracellular acidification rates (Figure 2I, p<0.05).

View Article: PubMed Central - PubMed

ABSTRACT

Here, we found that down-regulated expression of BTG3 might be positively correlated with colorectal carcinogenesis and its overexpression suppressed proliferation, glycolysis, mitochondrial respiration, cell cycle progression, migration, and invasion, and induced apoptosis, senescence and differentiation in SW480 and SW620 cells. After treated with cisplatin, MG132, paclitaxel and SAHA, BTG3 transfectants exhibited lower viability and higher apoptosis than the control in both time- and dose-dependent manners. BTG3 overexpression up- regulated the protein expression of Cyclin E, p16, p27, NF-&kappa;B, p38&alpha;/&beta;, XIAP, Bcl-2, ATG14 and p53, but down-regulated the mRNA expression of MRP1, BCRP, and mTOR in SW480 and SW620 cells. BTG3 overexpression inhibited tumor growth of SW620 cells by suppressing proliferation and inducing apoptosis. It was suggested that down-regulated BTG3 expression might be considered as a marker for colorectal carcinogenesis. BTG3 overexpression might reverse the aggressive phenotypes and be employed as a potential target for gene therapy of colorectal cancer.

No MeSH data available.


Related in: MedlinePlus