Long non-coding RNA RP11-480I12.5 promotes cervical carcinoma progression by regulating the Wnt/β-catenin signaling pathway

Abstract

The long non-coding RNA (lncRNA), RP11-480I12.5 is one of the most dysregulated lncRNAs, which is believed to contribute to the progression of cervical carcinoma (CC); however, the exact function of RP11-480I12.5 in human CC remains unknown. The present study aimed to investigate the function and underlying molecular mechanism of RP11-480I12.5 in CC. First, reverse transcription-quantitative PCR was implemented in order to detect differences in the expression of RP11-480I12.5 between normal and CC tissues. The present study used in vitro analysis to establish RP11-480I12.5 stable knockdown and overexpressing cell lines, in order to investigate the function and potential molecular mechanism of RP11-480I12.5 in the progression of CC. RP11-480I12.5 was upregulated in CC tissue compared with normal tissue. Furthermore, RP11-480I12.5 was associated with clinical stage, tumor size and lymph node metastasis. RP11-480I12.5 promoted the proliferation, migration and invasion of CC cell lines. Subsequently, the present study investigated the association between RP11-480I12.5 and the epithelial-to-mesenchymal transition (EMT) and Wnt/β-catenin pathways. RP11-480I12.5 promoted EMT through the Wnt/β-catenin pathway. Overall, the results of the present study demonstrate that RP11-480I12.5 promotes cercical cancer cell migration, invasion and EMT through the Wnt/β-catenin pathway.

Keywords: cervical cancer; epithelial-to-mesenchymal transition; long non-coding RNA; migration and invasion.

Figure 1.

RP11-480I12.5 is upregulated in CC tissues and negatively associated with prognosis. (A) Relative level of RP11-480I12.5 in TCGA database. (B) Relative level of RP11-480I12.5 in the in-house database containing 50 cases (two-tailed paired Student's t-test, (***P<0.001). (C) Relative level of RP11-480I12.5 at different stages of CC. One-way ANOVA test followed by Tukey's post hoc test, *P<0.05; **P<0.01; ***P<0.001. (D) Survival analysis in TCGA database. (E) Survival analysis in the in-house database containing 50 cases. CC, cervical cancer; TCGA, The Cancer Genome Atlas; N, non-cancerous; T, tumor; TPM, Transcripts PerKilobase Million; HR, hazard ratio.

Figure 2.

RP11-480I12.5 promotes the proliferation of CC. (A) Relative level of RP11-480I12.5 in the CC and normal cell lines. (B) Relative level of RP11-480I12.5 in cell lines. (C) Absorbance at 450 nm in the different cell lines. (D) Representative image from the colony formation assay. (E) Colony index of the indicated cells. (F) Representative image of Edu in different cell lines (magnification, ×200). (G) Statistical analysis of Edu⁺ cell lines. One-way ANOVA test followed by Tukey's post hoc test, *P<0.05; **P<0.01 vs. NC. CC, cervical cancer; sh, short hairpin, OV, overexpression; NC, negative control.

Figure 3.

RP11-480I12.5 promotes the migration and invasion of CC. (A) Representative images of wound healing assay (Scale bar, 50 µm). (B) Statistical analysis of wound healing. (C) Representative image of the Transwell and invasion chamber assays. (D) Statistical analysis of the Transwell and invasion chamber assays. One-way ANOVA test followed by Tukey's post hoc test, **P<0.01 vs. NC. CC, cervical cancer; NC, negative control; sh, short hairpin, OV, overexpression.

Figure 4.

RP11-480I12.5 induces EMT in CC through the Wnt/β-catenin pathway. (A) Western blot analysis of EMT markers and β-catenin. (B) Relative RNA levels of EMT markers and β-catenin. One-way ANOVA test followed by Tukey's post hoc test, **P<0.01 vs. NC. EMT; epithelial-to-mesenchymal transition; CC, cervical cancer; NC, negative control; sh, short hairpin; OV, overexpression.