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MicroRNA-638 inhibits cell proliferation by targeting phospholipase D1 in human gastric carcinoma.

Zhang J, Bian Z, Zhou J, Song M, Liu Z, Feng Y, Zhe L, Zhang B, Yin Y, Huang Z - Protein Cell (2015)

Bottom Line: From this research, we found that miR-638 was downregulated in GC tissues compared with corresponding noncancerous tissues (NCTs), and the DNA copy number of miR-638 was lower in GC than NCTs, which may induce the corresponding downregulation of miR-638 in GC.Subsequently, we identified that PLD1 is the target gene of miR-638 in GC, and silencing PLD1 expression phenocopied the inhibitory effect of miR-638 on GC cell proliferation.In summary, we revealed that miR-638 functions as a tumor suppressor in GC through inhibiting PLD1.

View Article: PubMed Central - PubMed

Affiliation: Wuxi Oncology Institute, The Affiliated Hospital of Jiangnan University, Wuxi, 214062, China.

ABSTRACT
MicroRNAs (miRNAs) are a type of small non-coding RNAs that are often play important roles in carcinogenesis, but the carcinogenic mechanism of miRNAs is still unclear. This study will investigate the function and the mechanism of miR-638 in carcinoma (GC). The expression of miR-638 in GC and the DNA copy number of miR-638 were detected by real-time PCR. The effect of miR-638 on cell proliferation was measured by counting kit-8 assay. Different assays, including bioinformatics algorithms (TargetScan and miRanda), luciferase report assay and Western blotting, were used to identify the target gene of miR-638 in GC. The expression of miR-638 target gene in clinical CRC tissues was also validated by immunohistochemical assay. From this research, we found that miR-638 was downregulated in GC tissues compared with corresponding noncancerous tissues (NCTs), and the DNA copy number of miR-638 was lower in GC than NCTs, which may induce the corresponding downregulation of miR-638 in GC. Ectopic expression of miR-638 inhibited GC cell growth in vitro. Subsequently, we identified that PLD1 is the target gene of miR-638 in GC, and silencing PLD1 expression phenocopied the inhibitory effect of miR-638 on GC cell proliferation. Furthermore, we observed that PLD1 was overexpressed in GC tissues, and high expression of PLD1 in GC predicted poor overall survival. In summary, we revealed that miR-638 functions as a tumor suppressor in GC through inhibiting PLD1.

No MeSH data available.


Related in: MedlinePlus

MiR-638 repressed GC cell proliferation by inhibiting PLD1. (A) PLD1 knockdown repressed MKN-45 and SGC-7901 cells growth, whereas upregulation of miR-638 in PLD1-depleted cells did not repress cell proliferation further. (B) MiR-638 silencing promoted cell growth, but did not promote cell proliferation in PLD1-depleted GC cells. (C) The upregulation of PLD1 ORF markedly promoted cell growth and abrogated miR-638-induced cell growth inhibition in MKN-45 and SGC-7901 cells. (D) MiR-638 overexpression promoted apoptosis, PLD1 overexpression repressed cell apoptosis and decreased the percentage of apoptotic cells after miR-638 overexpression. (E) The protein level of PLD1 was measured by Western blotting. Significant differences are indicated with * (*P < 0.05; **P < 0.01)
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Fig4: MiR-638 repressed GC cell proliferation by inhibiting PLD1. (A) PLD1 knockdown repressed MKN-45 and SGC-7901 cells growth, whereas upregulation of miR-638 in PLD1-depleted cells did not repress cell proliferation further. (B) MiR-638 silencing promoted cell growth, but did not promote cell proliferation in PLD1-depleted GC cells. (C) The upregulation of PLD1 ORF markedly promoted cell growth and abrogated miR-638-induced cell growth inhibition in MKN-45 and SGC-7901 cells. (D) MiR-638 overexpression promoted apoptosis, PLD1 overexpression repressed cell apoptosis and decreased the percentage of apoptotic cells after miR-638 overexpression. (E) The protein level of PLD1 was measured by Western blotting. Significant differences are indicated with * (*P < 0.05; **P < 0.01)

Mentions: To further examine whether PLD1 is a direct functional target of miR-638 in GC cells, we performed a series of functional restoration assays. As shown in Fig. 4A, the proliferation of MKN-45 and SGC-7901 cells transfected with si-PLD1 were significantly decreased (Fig. 4A). Also, in miR-638 transfection group, the proliferation ability of GC cells were decreased, whereas anti-miR-638 could not restore cell proliferation in PLD1-knockdown GC cells (Fig. 4B). Furthermore, we revealed that PLD1 over-expression could significantly promoted cell proliferation, which could not be repressed by either the exogenous or endogenous overexpression of miR-638 (Fig. 4C–E). Taken together, all of these results proved that miR-638 inhibits cell proliferation via directly targeting PLD1.Figure 4


MicroRNA-638 inhibits cell proliferation by targeting phospholipase D1 in human gastric carcinoma.

Zhang J, Bian Z, Zhou J, Song M, Liu Z, Feng Y, Zhe L, Zhang B, Yin Y, Huang Z - Protein Cell (2015)

MiR-638 repressed GC cell proliferation by inhibiting PLD1. (A) PLD1 knockdown repressed MKN-45 and SGC-7901 cells growth, whereas upregulation of miR-638 in PLD1-depleted cells did not repress cell proliferation further. (B) MiR-638 silencing promoted cell growth, but did not promote cell proliferation in PLD1-depleted GC cells. (C) The upregulation of PLD1 ORF markedly promoted cell growth and abrogated miR-638-induced cell growth inhibition in MKN-45 and SGC-7901 cells. (D) MiR-638 overexpression promoted apoptosis, PLD1 overexpression repressed cell apoptosis and decreased the percentage of apoptotic cells after miR-638 overexpression. (E) The protein level of PLD1 was measured by Western blotting. Significant differences are indicated with * (*P < 0.05; **P < 0.01)
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Related In: Results  -  Collection

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Fig4: MiR-638 repressed GC cell proliferation by inhibiting PLD1. (A) PLD1 knockdown repressed MKN-45 and SGC-7901 cells growth, whereas upregulation of miR-638 in PLD1-depleted cells did not repress cell proliferation further. (B) MiR-638 silencing promoted cell growth, but did not promote cell proliferation in PLD1-depleted GC cells. (C) The upregulation of PLD1 ORF markedly promoted cell growth and abrogated miR-638-induced cell growth inhibition in MKN-45 and SGC-7901 cells. (D) MiR-638 overexpression promoted apoptosis, PLD1 overexpression repressed cell apoptosis and decreased the percentage of apoptotic cells after miR-638 overexpression. (E) The protein level of PLD1 was measured by Western blotting. Significant differences are indicated with * (*P < 0.05; **P < 0.01)
Mentions: To further examine whether PLD1 is a direct functional target of miR-638 in GC cells, we performed a series of functional restoration assays. As shown in Fig. 4A, the proliferation of MKN-45 and SGC-7901 cells transfected with si-PLD1 were significantly decreased (Fig. 4A). Also, in miR-638 transfection group, the proliferation ability of GC cells were decreased, whereas anti-miR-638 could not restore cell proliferation in PLD1-knockdown GC cells (Fig. 4B). Furthermore, we revealed that PLD1 over-expression could significantly promoted cell proliferation, which could not be repressed by either the exogenous or endogenous overexpression of miR-638 (Fig. 4C–E). Taken together, all of these results proved that miR-638 inhibits cell proliferation via directly targeting PLD1.Figure 4

Bottom Line: From this research, we found that miR-638 was downregulated in GC tissues compared with corresponding noncancerous tissues (NCTs), and the DNA copy number of miR-638 was lower in GC than NCTs, which may induce the corresponding downregulation of miR-638 in GC.Subsequently, we identified that PLD1 is the target gene of miR-638 in GC, and silencing PLD1 expression phenocopied the inhibitory effect of miR-638 on GC cell proliferation.In summary, we revealed that miR-638 functions as a tumor suppressor in GC through inhibiting PLD1.

View Article: PubMed Central - PubMed

Affiliation: Wuxi Oncology Institute, The Affiliated Hospital of Jiangnan University, Wuxi, 214062, China.

ABSTRACT
MicroRNAs (miRNAs) are a type of small non-coding RNAs that are often play important roles in carcinogenesis, but the carcinogenic mechanism of miRNAs is still unclear. This study will investigate the function and the mechanism of miR-638 in carcinoma (GC). The expression of miR-638 in GC and the DNA copy number of miR-638 were detected by real-time PCR. The effect of miR-638 on cell proliferation was measured by counting kit-8 assay. Different assays, including bioinformatics algorithms (TargetScan and miRanda), luciferase report assay and Western blotting, were used to identify the target gene of miR-638 in GC. The expression of miR-638 target gene in clinical CRC tissues was also validated by immunohistochemical assay. From this research, we found that miR-638 was downregulated in GC tissues compared with corresponding noncancerous tissues (NCTs), and the DNA copy number of miR-638 was lower in GC than NCTs, which may induce the corresponding downregulation of miR-638 in GC. Ectopic expression of miR-638 inhibited GC cell growth in vitro. Subsequently, we identified that PLD1 is the target gene of miR-638 in GC, and silencing PLD1 expression phenocopied the inhibitory effect of miR-638 on GC cell proliferation. Furthermore, we observed that PLD1 was overexpressed in GC tissues, and high expression of PLD1 in GC predicted poor overall survival. In summary, we revealed that miR-638 functions as a tumor suppressor in GC through inhibiting PLD1.

No MeSH data available.


Related in: MedlinePlus