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MicroRNA-135a acts as a putative tumor suppressor by directly targeting very low density lipoprotein receptor in human gallbladder cancer.

Zhou H, Guo W, Zhao Y, Wang Y, Zha R, Ding J, Liang L, Yang G, Chen Z, Ma B, Yin B - Cancer Sci. (2014)

Bottom Line: Moreover, very low density lipoprotein receptor (VLDLR), which is often upregulated in GBC tissues, was identified as a direct functional target of miR-135a-5p.Furthermore, the p38 MAPK pathway was proven to be involved in miR-135a-VLDLR downstream signaling.Together, these results suggested that the miR-135a-VLDLR-p38 axis may contribute to GBC cell proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, Huashan Hospital, Fudan University, Shanghai, China.

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Related in: MedlinePlus

MicroRNA-135a (miR-135a) downregulates very low density lipoprotein receptor (VLDLR) expression by directly targeting its 3′-UTR. (a) Search for the identification of potential genes combining microarray assays, TargetScan prediction, and PicTar prediction. (b) Potential target genes' mRNA levels in GBC-SD cells transfected with miR-135a-5p mimic or negative control (NC). (c) Putative binding site for miR-135a-5p within human (Has), chimpanzee (Ptr), and rhesus (Mml) VLDLR 3′-UTR. Binding site sequences are indicated with blue, the sequences of miR-135a-5p binding site in the wild-type (indicated with blue), and mutant (indicated with red) VLDLR 3′-UTR. (d) Relative luciferase activity analyses. The pLUC, pLUC-wild-type–VLDLR–3′-UTR, or pLUC-mutant–VLDLR–3′-UTR was transfected into HEK-293T cells with pWPXL or pWPXL-miR-135a. Renilla luciferase vector was used as an internal control. Representative experiments are shown with the mean ± SD. (e) Regression and correlation analysis of miR-135a-5p and VLDLR mRNA in tissues. (f) Protein levels of VLDLR were determined by Western blot assays in GBC-SD, EH-GB1, and SGC-996 cells transfected with miR-135a-5p mimic or NC. (g) The VLDLR protein level was often downregulated in tumor tissue (T). (N means non-tumor tissue).
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fig03: MicroRNA-135a (miR-135a) downregulates very low density lipoprotein receptor (VLDLR) expression by directly targeting its 3′-UTR. (a) Search for the identification of potential genes combining microarray assays, TargetScan prediction, and PicTar prediction. (b) Potential target genes' mRNA levels in GBC-SD cells transfected with miR-135a-5p mimic or negative control (NC). (c) Putative binding site for miR-135a-5p within human (Has), chimpanzee (Ptr), and rhesus (Mml) VLDLR 3′-UTR. Binding site sequences are indicated with blue, the sequences of miR-135a-5p binding site in the wild-type (indicated with blue), and mutant (indicated with red) VLDLR 3′-UTR. (d) Relative luciferase activity analyses. The pLUC, pLUC-wild-type–VLDLR–3′-UTR, or pLUC-mutant–VLDLR–3′-UTR was transfected into HEK-293T cells with pWPXL or pWPXL-miR-135a. Renilla luciferase vector was used as an internal control. Representative experiments are shown with the mean ± SD. (e) Regression and correlation analysis of miR-135a-5p and VLDLR mRNA in tissues. (f) Protein levels of VLDLR were determined by Western blot assays in GBC-SD, EH-GB1, and SGC-996 cells transfected with miR-135a-5p mimic or NC. (g) The VLDLR protein level was often downregulated in tumor tissue (T). (N means non-tumor tissue).

Mentions: Potential targets of miR-135a-5p were predicted by TargetScan (http://www.targetscan.org) and PicTar (http://pictar.mdc-berlin.de), and mRNA microarray assays were carried out in EH-GB1 cells transfected with miR-135a-5p mimic. Six genes were considered as the candidate target gene (Fig. 3a). In GBC-SD cells, miR-135a-5p mimic could dampen the mRNA levels of SP3 and VLDLR, which have inhibitory effects on cancer cell proliferation (Fig. 3b).(11,12) Very low density lipoprotein receptor siRNAs could clearly inhibit the proliferation of both EH-GB1 and GBC-SD cells, yet SP3 siRNAs did not affect GBC-SD cell proliferation (Fig. S4), indicating that VLDLR was responsible for the effect of miR-135a-5p on GBC cell proliferation.


MicroRNA-135a acts as a putative tumor suppressor by directly targeting very low density lipoprotein receptor in human gallbladder cancer.

Zhou H, Guo W, Zhao Y, Wang Y, Zha R, Ding J, Liang L, Yang G, Chen Z, Ma B, Yin B - Cancer Sci. (2014)

MicroRNA-135a (miR-135a) downregulates very low density lipoprotein receptor (VLDLR) expression by directly targeting its 3′-UTR. (a) Search for the identification of potential genes combining microarray assays, TargetScan prediction, and PicTar prediction. (b) Potential target genes' mRNA levels in GBC-SD cells transfected with miR-135a-5p mimic or negative control (NC). (c) Putative binding site for miR-135a-5p within human (Has), chimpanzee (Ptr), and rhesus (Mml) VLDLR 3′-UTR. Binding site sequences are indicated with blue, the sequences of miR-135a-5p binding site in the wild-type (indicated with blue), and mutant (indicated with red) VLDLR 3′-UTR. (d) Relative luciferase activity analyses. The pLUC, pLUC-wild-type–VLDLR–3′-UTR, or pLUC-mutant–VLDLR–3′-UTR was transfected into HEK-293T cells with pWPXL or pWPXL-miR-135a. Renilla luciferase vector was used as an internal control. Representative experiments are shown with the mean ± SD. (e) Regression and correlation analysis of miR-135a-5p and VLDLR mRNA in tissues. (f) Protein levels of VLDLR were determined by Western blot assays in GBC-SD, EH-GB1, and SGC-996 cells transfected with miR-135a-5p mimic or NC. (g) The VLDLR protein level was often downregulated in tumor tissue (T). (N means non-tumor tissue).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig03: MicroRNA-135a (miR-135a) downregulates very low density lipoprotein receptor (VLDLR) expression by directly targeting its 3′-UTR. (a) Search for the identification of potential genes combining microarray assays, TargetScan prediction, and PicTar prediction. (b) Potential target genes' mRNA levels in GBC-SD cells transfected with miR-135a-5p mimic or negative control (NC). (c) Putative binding site for miR-135a-5p within human (Has), chimpanzee (Ptr), and rhesus (Mml) VLDLR 3′-UTR. Binding site sequences are indicated with blue, the sequences of miR-135a-5p binding site in the wild-type (indicated with blue), and mutant (indicated with red) VLDLR 3′-UTR. (d) Relative luciferase activity analyses. The pLUC, pLUC-wild-type–VLDLR–3′-UTR, or pLUC-mutant–VLDLR–3′-UTR was transfected into HEK-293T cells with pWPXL or pWPXL-miR-135a. Renilla luciferase vector was used as an internal control. Representative experiments are shown with the mean ± SD. (e) Regression and correlation analysis of miR-135a-5p and VLDLR mRNA in tissues. (f) Protein levels of VLDLR were determined by Western blot assays in GBC-SD, EH-GB1, and SGC-996 cells transfected with miR-135a-5p mimic or NC. (g) The VLDLR protein level was often downregulated in tumor tissue (T). (N means non-tumor tissue).
Mentions: Potential targets of miR-135a-5p were predicted by TargetScan (http://www.targetscan.org) and PicTar (http://pictar.mdc-berlin.de), and mRNA microarray assays were carried out in EH-GB1 cells transfected with miR-135a-5p mimic. Six genes were considered as the candidate target gene (Fig. 3a). In GBC-SD cells, miR-135a-5p mimic could dampen the mRNA levels of SP3 and VLDLR, which have inhibitory effects on cancer cell proliferation (Fig. 3b).(11,12) Very low density lipoprotein receptor siRNAs could clearly inhibit the proliferation of both EH-GB1 and GBC-SD cells, yet SP3 siRNAs did not affect GBC-SD cell proliferation (Fig. S4), indicating that VLDLR was responsible for the effect of miR-135a-5p on GBC cell proliferation.

Bottom Line: Moreover, very low density lipoprotein receptor (VLDLR), which is often upregulated in GBC tissues, was identified as a direct functional target of miR-135a-5p.Furthermore, the p38 MAPK pathway was proven to be involved in miR-135a-VLDLR downstream signaling.Together, these results suggested that the miR-135a-VLDLR-p38 axis may contribute to GBC cell proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, Huashan Hospital, Fudan University, Shanghai, China.

Show MeSH
Related in: MedlinePlus