MicroRNA-148b suppresses proliferation, migration, and invasion of nasopharyngeal carcinoma cells by targeting metastasis-associated gene 2

Figures & data

Figure 1 Downregulation of miR-148b in NPC tissues and cell lines.

Notes: (A) Expression of miR-148b in 20 cases of noncancerous NP tissues and 25 cases of NPC tissues was detected by qRT-PCR. (B) Expression of miR-148b in NPC cell lines (CNE1, CNE2, C666-1 and HNE1) and nasopharyngeal epithelium cell, NP69, was detected by qRT-PCR (*P<0.05 vs NP69).
Abbreviations: miR, microRNA; NP, nasopharyngeal; NPC, nasopharyngeal carcinoma; qRT-PCR, quantitative reverse transcription polymerase chain reaction; U6, U6 snRNA.

Figure 2 Inhibition effects of miR-148b on proliferation, migration, and invasion of CNE2 and C666-1 cells.

Notes: (A) Forty-eight hours after transfection with miR-148b mimic or miR-Ctrl. The miR-148b expression in CNE2 and C666-1 cells was determined by qRT-PCR. Colony formation assays (B) and CCK-8 assays (C) were conducted to evaluate the effects of miR-148b overexpression on proliferation of CNE2 and C666-1 cells. (D) Transwell assays were conducted to evaluate the effects of miR-148b on migration and invasion of CNE2 and C666-1 cells (*P<0.05, ** P<0.01).
Abbreviations: CCK-8, Cell Counting Kit-8; Ctrl, control; h, hours; miR, microRNA; NS, no significance; qRT-PCR, quantitative reverse transcription polymerase chain reaction.

Figure 3 MTA2 is a target of miR-148b.

Notes: (A) The binding site of miR-148b in the 395–401 bp position in the 3′UTR of MTA2 mRNA predicted by TargetScan. (B) The dual luciferase reporter activity analysis of WT and Mut type MTA2 3′UTR reporter constructs after co-transfection with miR-148b or miR-Ctrl. (C) Ectopic miR-148b expression reduced the protein expression of MTA2 in CNE2 and C666-1 cells. (D) The correlation between miR-148b and MTA2 mRNA expression in 25 cases of NPC tissue was analyzed by Pearson’s correlation assay (**P<0.01).
Abbreviations: Ctrl, control; miR, microRNA; MTA2, metastasis-associated gene 2; Mut, mutant; NPC, nasopharyngeal carcinoma; NS, no significance; WT, wild type; UTR, untranslated region.

Figure 4 The tumor-suppression effects of miR-148b on C666-1 cell were partially reversed by MTA2 restoration.

Notes: (A) C666-1 cells were transfected with the empty vector or MTA2 expression vector in miR-148b-overexpressing C666-1 cells, and MTA2 protein expression was analyzed by Western blot. The effects of MTA2 restoration on cell growth were detected by colony formation assays (B) and CCK-8 assays (C). (D) The effects of MTA2 restoration on cell migration and invasion were detected by transwell assays (*P<0.05, **P<0.01).
Abbreviations: CCK-8, Cell Counting Kit-8; Ctrl, control; miR, microRNA; h, hours; MTA2, metastasis-associated gene 2; pcMTA2, MTA2 expression plasmid pcDNA3.1.