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MicroRNA-140 Inhibits Cell Proliferation in Gastric Cancer Cell Line HGC-27 by Suppressing SOX4.

Zou J, Xu Y - Med. Sci. Monit. (2016)

Bottom Line: SOX4, a predicted target of miR-140, was mutated to verify its regulation by miR-140, and was overexpressed to analyze its function in cell proliferation.Doxorubicin treatment was performed to investigate the effect of miR-140 on drug resistance.Overexpressing SOX4 led to promoted cell viability, colony formation, and cell cycle progress. miR-140 overexpression also improved the anti-viability effects of doxorubicin, suggesting its potential in reducing the drug resistance of gastric cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (mainland).

ABSTRACT
BACKGROUND Gastric cancer is a malignant tumor with a high morbidity and mortality. MicroRNAs are important regulators of gene expression, influencing the progression of gastric cancer. This study aimed to reveal the role of microRNA-140 (miR-140) in gastric cancer cell proliferation and its potential mechanisms. MATERIAL AND METHODS Gastric cancer tissues and cell lines BGC-823, SGC-7901, and HGC-27 were used to analyze miR-140 levels compared to normal tissues and cell line GES-1. In HGC-27 cells transfected with miR-140 mimic, we performed MTT, colony formation assay, and cell cycle assay by flow cytometry. SOX4, a predicted target of miR-140, was mutated to verify its regulation by miR-140, and was overexpressed to analyze its function in cell proliferation. Doxorubicin treatment was performed to investigate the effect of miR-140 on drug resistance. RESULTS miR-140 was down-regulated in gastric cancer tissues and cell lines, with the lowest expression level in HGC-27. miR-140 overexpression inhibited HGC-27 cell viability and colony formation and resulted in G0/G1 arrest. miR-140 suppressed SOX4 expression via binding to the 3' untranslated region, while the mutant SOX4 could not be regulated. Overexpressing SOX4 led to promoted cell viability, colony formation, and cell cycle progress. miR-140 overexpression also improved the anti-viability effects of doxorubicin, suggesting its potential in reducing the drug resistance of gastric cells. CONCLUSIONS These findings suggest that miR-140 directly inhibits SOX4, which might be one of its mechanisms in suppressing gastric cancer cell proliferation. This study provides a promising therapeutic strategy for treating gastric cancer and facilitates microRNA research in various diseases.

No MeSH data available.


Related in: MedlinePlus

SOX4 promotes HGC-27 cell viability, proliferation, and cell cycle process. (A) MTT data show that cell viability is promoted by SOX4 overexpression when detected at 24, 48, and 72 h after transfection. (B) Colony formation assay shows the colony is increased by SOX4 overexpression after 2 weeks of incubation. (C) Histogram reflecting the flow cytometry result indicates that the cell percent in the G0/G1 phase is decreased by SOX4 overexpression. (D) Western blot shows CCND1 and CDK2 protein levels are up-regulated by SOX4 overexpression. GAPDH is used as the internal control. * P<0.05. SOX4 – sex-determining region Y (SRY)-box 4. CCND1 – cyclin D1. CDK2 – cyclin-dependent kinase 2.
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f4-medscimonit-22-2243: SOX4 promotes HGC-27 cell viability, proliferation, and cell cycle process. (A) MTT data show that cell viability is promoted by SOX4 overexpression when detected at 24, 48, and 72 h after transfection. (B) Colony formation assay shows the colony is increased by SOX4 overexpression after 2 weeks of incubation. (C) Histogram reflecting the flow cytometry result indicates that the cell percent in the G0/G1 phase is decreased by SOX4 overexpression. (D) Western blot shows CCND1 and CDK2 protein levels are up-regulated by SOX4 overexpression. GAPDH is used as the internal control. * P<0.05. SOX4 – sex-determining region Y (SRY)-box 4. CCND1 – cyclin D1. CDK2 – cyclin-dependent kinase 2.

Mentions: Was the inhibition of SOX4 by miR-140 associated with HGC-27 cell proliferation? To answer this question, we performed SOX4 overexpression by transfecting its overexpression vector to investigate the changes in cell viability, proliferation, and cell cycle. Cell viability was analyzed by MTT method and results showed that SOX4 overexpression could significantly promote HGC-27 cell viability at 24, 48, and 72 h after transfection (P<0.05, Figure 4A). Similarly, overexpressing SOX4 also increased the number of colonies formed after 2 weeks of incubation (P<0.05, Figure 4B), indicating the promotion of cell proliferation. Further, the percentage of cells in the G0/G1 phase was decreased by SOX4 overexpression compared to the control group (Figure 4C), which was in accordance to the up-regulated CCND1 and CDK2 protein levels (Figure 4D), implying that SOX4 might help to promote the G1/S transition. Taken together, these data suggest the roles of SOX4 in promoting HGC-27 cell viability, proliferation, and cell cycle processes, which were consistent with the above results that the anti-proliferative miR-140 inhibited SOX4. Therefore, the inhibition of SOX4 might be a potential mechanism by which miR-140 suppresses cell proliferation.


MicroRNA-140 Inhibits Cell Proliferation in Gastric Cancer Cell Line HGC-27 by Suppressing SOX4.

Zou J, Xu Y - Med. Sci. Monit. (2016)

SOX4 promotes HGC-27 cell viability, proliferation, and cell cycle process. (A) MTT data show that cell viability is promoted by SOX4 overexpression when detected at 24, 48, and 72 h after transfection. (B) Colony formation assay shows the colony is increased by SOX4 overexpression after 2 weeks of incubation. (C) Histogram reflecting the flow cytometry result indicates that the cell percent in the G0/G1 phase is decreased by SOX4 overexpression. (D) Western blot shows CCND1 and CDK2 protein levels are up-regulated by SOX4 overexpression. GAPDH is used as the internal control. * P<0.05. SOX4 – sex-determining region Y (SRY)-box 4. CCND1 – cyclin D1. CDK2 – cyclin-dependent kinase 2.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4930272&req=5

f4-medscimonit-22-2243: SOX4 promotes HGC-27 cell viability, proliferation, and cell cycle process. (A) MTT data show that cell viability is promoted by SOX4 overexpression when detected at 24, 48, and 72 h after transfection. (B) Colony formation assay shows the colony is increased by SOX4 overexpression after 2 weeks of incubation. (C) Histogram reflecting the flow cytometry result indicates that the cell percent in the G0/G1 phase is decreased by SOX4 overexpression. (D) Western blot shows CCND1 and CDK2 protein levels are up-regulated by SOX4 overexpression. GAPDH is used as the internal control. * P<0.05. SOX4 – sex-determining region Y (SRY)-box 4. CCND1 – cyclin D1. CDK2 – cyclin-dependent kinase 2.
Mentions: Was the inhibition of SOX4 by miR-140 associated with HGC-27 cell proliferation? To answer this question, we performed SOX4 overexpression by transfecting its overexpression vector to investigate the changes in cell viability, proliferation, and cell cycle. Cell viability was analyzed by MTT method and results showed that SOX4 overexpression could significantly promote HGC-27 cell viability at 24, 48, and 72 h after transfection (P<0.05, Figure 4A). Similarly, overexpressing SOX4 also increased the number of colonies formed after 2 weeks of incubation (P<0.05, Figure 4B), indicating the promotion of cell proliferation. Further, the percentage of cells in the G0/G1 phase was decreased by SOX4 overexpression compared to the control group (Figure 4C), which was in accordance to the up-regulated CCND1 and CDK2 protein levels (Figure 4D), implying that SOX4 might help to promote the G1/S transition. Taken together, these data suggest the roles of SOX4 in promoting HGC-27 cell viability, proliferation, and cell cycle processes, which were consistent with the above results that the anti-proliferative miR-140 inhibited SOX4. Therefore, the inhibition of SOX4 might be a potential mechanism by which miR-140 suppresses cell proliferation.

Bottom Line: SOX4, a predicted target of miR-140, was mutated to verify its regulation by miR-140, and was overexpressed to analyze its function in cell proliferation.Doxorubicin treatment was performed to investigate the effect of miR-140 on drug resistance.Overexpressing SOX4 led to promoted cell viability, colony formation, and cell cycle progress. miR-140 overexpression also improved the anti-viability effects of doxorubicin, suggesting its potential in reducing the drug resistance of gastric cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (mainland).

ABSTRACT
BACKGROUND Gastric cancer is a malignant tumor with a high morbidity and mortality. MicroRNAs are important regulators of gene expression, influencing the progression of gastric cancer. This study aimed to reveal the role of microRNA-140 (miR-140) in gastric cancer cell proliferation and its potential mechanisms. MATERIAL AND METHODS Gastric cancer tissues and cell lines BGC-823, SGC-7901, and HGC-27 were used to analyze miR-140 levels compared to normal tissues and cell line GES-1. In HGC-27 cells transfected with miR-140 mimic, we performed MTT, colony formation assay, and cell cycle assay by flow cytometry. SOX4, a predicted target of miR-140, was mutated to verify its regulation by miR-140, and was overexpressed to analyze its function in cell proliferation. Doxorubicin treatment was performed to investigate the effect of miR-140 on drug resistance. RESULTS miR-140 was down-regulated in gastric cancer tissues and cell lines, with the lowest expression level in HGC-27. miR-140 overexpression inhibited HGC-27 cell viability and colony formation and resulted in G0/G1 arrest. miR-140 suppressed SOX4 expression via binding to the 3' untranslated region, while the mutant SOX4 could not be regulated. Overexpressing SOX4 led to promoted cell viability, colony formation, and cell cycle progress. miR-140 overexpression also improved the anti-viability effects of doxorubicin, suggesting its potential in reducing the drug resistance of gastric cells. CONCLUSIONS These findings suggest that miR-140 directly inhibits SOX4, which might be one of its mechanisms in suppressing gastric cancer cell proliferation. This study provides a promising therapeutic strategy for treating gastric cancer and facilitates microRNA research in various diseases.

No MeSH data available.


Related in: MedlinePlus