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The SOX17/miR-371-5p/SOX2 axis inhibits EMT, stem cell properties and metastasis in colorectal cancer.

Li Y, Lv Z, He G, Wang J, Zhang X, Lu G, Ren X, Wang F, Zhu X, Ding Y, Liao W, Ding Y, Liang L - Oncotarget (2015)

Bottom Line: In the study, miR-371-5p was obviously down-regulated in primary CRC tissues compared with matched adjacent normal mucosa and correlated significantly with differentiation, tumor size, lymphatic and liver metastases.It also suppressed EMT by regulating Wnt/β-catenin signaling and strongly decreased the CRC stemness phenotypes.A positive relationship between SOX17 and miR-371-5p expression and a negative one between miR-371-5p and SOX2 expression were observed in CRC cell lines and tissues.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong Province, People's Republic of China.

ABSTRACT
Cancer stem cells (CSCs) and EMT-type cells, which share molecular characteristics with CSCs, have been believed to play critical roles in tumor metastasis. Although much progress has been garnered in elucidating the molecular pathways that trigger EMT, stemness and metastasis, a number of key mechanistic gaps remain elusive. In the study, miR-371-5p was obviously down-regulated in primary CRC tissues compared with matched adjacent normal mucosa and correlated significantly with differentiation, tumor size, lymphatic and liver metastases. MiR-371-5p could attenuate proliferation, invasion in vitro and metastasis in vivo in CRC cells. It also suppressed EMT by regulating Wnt/β-catenin signaling and strongly decreased the CRC stemness phenotypes. Moreover, demethylation of SOX17 induced miR-371-5p expression and consequently suppressed its direct target SOX2 in CRC cells. MiR-371-5p was necessary for SOX17 mediated cancer-related traits and SOX2 was a functional target of miR-371-5p. A positive relationship between SOX17 and miR-371-5p expression and a negative one between miR-371-5p and SOX2 expression were observed in CRC cell lines and tissues. In conclusion, we identified miR-371-5p as an important "oncosuppressor" in CRC progression and elucidated a novel mechanism of the SOX17/miR-371-5p/SOX2 axis in the regulation of EMT, stemness and metastasis, which may be a potential therapeutic target.

No MeSH data available.


Related in: MedlinePlus

miR-371-5p is associated with CRC metastasis and suppresses invasion and EMT of CRC cells in vitro(A) Endogenous expression of miR-371-5p in 6 CRC cell lines and normal colon mucosa (N) by qRT-PCR. The relative expression levels of miR-371-5p in normal colon mucosa were normalized to 1. (B) Expression of miR-371-5p in 100 cases of the primary CRC tissues with or without metastasis and matched adjacent normal mucosa by qRT-PCR. The relative expression levels of miR-371-5p in normal mucosa were normalized to 1. (C) Effect of miR-371-5p ectopic expression or miR-371-5p knockdown on the invasiveness of CRC cells by Boyden chamber. Scale bars represent 20 μm. (D) Expression of EMT related markers and target genes of Wnt/β-catenin signaling in miR-371-5p over-expressing or depleting cells by Western blot. Expression levels were normalized to Tubulin. (E) Immunofluorescence images of E-cadherin, Vimentin expression and nuclear translocation of β-catenin in miR-371-5p depleting cells. Red scale bars represent 10 μm, whereas yellow scale bars represent 5 μm. * P < 0.05, ** P < 0.01. Data represent the mean ± SD.
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Figure 1: miR-371-5p is associated with CRC metastasis and suppresses invasion and EMT of CRC cells in vitro(A) Endogenous expression of miR-371-5p in 6 CRC cell lines and normal colon mucosa (N) by qRT-PCR. The relative expression levels of miR-371-5p in normal colon mucosa were normalized to 1. (B) Expression of miR-371-5p in 100 cases of the primary CRC tissues with or without metastasis and matched adjacent normal mucosa by qRT-PCR. The relative expression levels of miR-371-5p in normal mucosa were normalized to 1. (C) Effect of miR-371-5p ectopic expression or miR-371-5p knockdown on the invasiveness of CRC cells by Boyden chamber. Scale bars represent 20 μm. (D) Expression of EMT related markers and target genes of Wnt/β-catenin signaling in miR-371-5p over-expressing or depleting cells by Western blot. Expression levels were normalized to Tubulin. (E) Immunofluorescence images of E-cadherin, Vimentin expression and nuclear translocation of β-catenin in miR-371-5p depleting cells. Red scale bars represent 10 μm, whereas yellow scale bars represent 5 μm. * P < 0.05, ** P < 0.01. Data represent the mean ± SD.

Mentions: To investigate the expression pattern and clinicopathologic significance of miR-371-5p in CRC, we first detected miR-371-5p expression in 6 CRC cell lines with different metastatic potentials. Real-time PCR analyses showed that miR-371-5p was obviously down-regulated in 6 CRC cell lines compared with normal colon mucosa. The expression levels of miR-371-5p were lower in high metastatic SW620 and Lovo cell lines than those with low metastatic abilities (Figure 1A). Thus we examined the expression and clinical values of miR-371-5p in 100 cases of paired CRC tissues. The expression of miR-371-5p was obviously lower in primary CRC tissues than in matched adjacent non-tumor mucosa. Strikingly, miR-371-5p was markedly down-regulated in primary CRC tissues with metastasis compared with those without metastasis (Figure 1B). Moreover, miR-371-5p expression correlated significantly with differentiation, tumor size, lymphatic and liver metastases (p < 0.05; Supplementary Table 1). Since miR-371-5p and miR-371-3p are derived from a single precursor, we also assessed the expression of miR-371-3p in CRC tissues. However, there was no significant difference of miR-371-3p expression between primary CRC tissues and matched adjacent normal mucosa (Supplementary Figure 1A). The above results suggest a possible link between down-regulation of miR-371-5p and CRC metastasis.


The SOX17/miR-371-5p/SOX2 axis inhibits EMT, stem cell properties and metastasis in colorectal cancer.

Li Y, Lv Z, He G, Wang J, Zhang X, Lu G, Ren X, Wang F, Zhu X, Ding Y, Liao W, Ding Y, Liang L - Oncotarget (2015)

miR-371-5p is associated with CRC metastasis and suppresses invasion and EMT of CRC cells in vitro(A) Endogenous expression of miR-371-5p in 6 CRC cell lines and normal colon mucosa (N) by qRT-PCR. The relative expression levels of miR-371-5p in normal colon mucosa were normalized to 1. (B) Expression of miR-371-5p in 100 cases of the primary CRC tissues with or without metastasis and matched adjacent normal mucosa by qRT-PCR. The relative expression levels of miR-371-5p in normal mucosa were normalized to 1. (C) Effect of miR-371-5p ectopic expression or miR-371-5p knockdown on the invasiveness of CRC cells by Boyden chamber. Scale bars represent 20 μm. (D) Expression of EMT related markers and target genes of Wnt/β-catenin signaling in miR-371-5p over-expressing or depleting cells by Western blot. Expression levels were normalized to Tubulin. (E) Immunofluorescence images of E-cadherin, Vimentin expression and nuclear translocation of β-catenin in miR-371-5p depleting cells. Red scale bars represent 10 μm, whereas yellow scale bars represent 5 μm. * P < 0.05, ** P < 0.01. Data represent the mean ± SD.
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Figure 1: miR-371-5p is associated with CRC metastasis and suppresses invasion and EMT of CRC cells in vitro(A) Endogenous expression of miR-371-5p in 6 CRC cell lines and normal colon mucosa (N) by qRT-PCR. The relative expression levels of miR-371-5p in normal colon mucosa were normalized to 1. (B) Expression of miR-371-5p in 100 cases of the primary CRC tissues with or without metastasis and matched adjacent normal mucosa by qRT-PCR. The relative expression levels of miR-371-5p in normal mucosa were normalized to 1. (C) Effect of miR-371-5p ectopic expression or miR-371-5p knockdown on the invasiveness of CRC cells by Boyden chamber. Scale bars represent 20 μm. (D) Expression of EMT related markers and target genes of Wnt/β-catenin signaling in miR-371-5p over-expressing or depleting cells by Western blot. Expression levels were normalized to Tubulin. (E) Immunofluorescence images of E-cadherin, Vimentin expression and nuclear translocation of β-catenin in miR-371-5p depleting cells. Red scale bars represent 10 μm, whereas yellow scale bars represent 5 μm. * P < 0.05, ** P < 0.01. Data represent the mean ± SD.
Mentions: To investigate the expression pattern and clinicopathologic significance of miR-371-5p in CRC, we first detected miR-371-5p expression in 6 CRC cell lines with different metastatic potentials. Real-time PCR analyses showed that miR-371-5p was obviously down-regulated in 6 CRC cell lines compared with normal colon mucosa. The expression levels of miR-371-5p were lower in high metastatic SW620 and Lovo cell lines than those with low metastatic abilities (Figure 1A). Thus we examined the expression and clinical values of miR-371-5p in 100 cases of paired CRC tissues. The expression of miR-371-5p was obviously lower in primary CRC tissues than in matched adjacent non-tumor mucosa. Strikingly, miR-371-5p was markedly down-regulated in primary CRC tissues with metastasis compared with those without metastasis (Figure 1B). Moreover, miR-371-5p expression correlated significantly with differentiation, tumor size, lymphatic and liver metastases (p < 0.05; Supplementary Table 1). Since miR-371-5p and miR-371-3p are derived from a single precursor, we also assessed the expression of miR-371-3p in CRC tissues. However, there was no significant difference of miR-371-3p expression between primary CRC tissues and matched adjacent normal mucosa (Supplementary Figure 1A). The above results suggest a possible link between down-regulation of miR-371-5p and CRC metastasis.

Bottom Line: In the study, miR-371-5p was obviously down-regulated in primary CRC tissues compared with matched adjacent normal mucosa and correlated significantly with differentiation, tumor size, lymphatic and liver metastases.It also suppressed EMT by regulating Wnt/β-catenin signaling and strongly decreased the CRC stemness phenotypes.A positive relationship between SOX17 and miR-371-5p expression and a negative one between miR-371-5p and SOX2 expression were observed in CRC cell lines and tissues.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong Province, People's Republic of China.

ABSTRACT
Cancer stem cells (CSCs) and EMT-type cells, which share molecular characteristics with CSCs, have been believed to play critical roles in tumor metastasis. Although much progress has been garnered in elucidating the molecular pathways that trigger EMT, stemness and metastasis, a number of key mechanistic gaps remain elusive. In the study, miR-371-5p was obviously down-regulated in primary CRC tissues compared with matched adjacent normal mucosa and correlated significantly with differentiation, tumor size, lymphatic and liver metastases. MiR-371-5p could attenuate proliferation, invasion in vitro and metastasis in vivo in CRC cells. It also suppressed EMT by regulating Wnt/β-catenin signaling and strongly decreased the CRC stemness phenotypes. Moreover, demethylation of SOX17 induced miR-371-5p expression and consequently suppressed its direct target SOX2 in CRC cells. MiR-371-5p was necessary for SOX17 mediated cancer-related traits and SOX2 was a functional target of miR-371-5p. A positive relationship between SOX17 and miR-371-5p expression and a negative one between miR-371-5p and SOX2 expression were observed in CRC cell lines and tissues. In conclusion, we identified miR-371-5p as an important "oncosuppressor" in CRC progression and elucidated a novel mechanism of the SOX17/miR-371-5p/SOX2 axis in the regulation of EMT, stemness and metastasis, which may be a potential therapeutic target.

No MeSH data available.


Related in: MedlinePlus