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Expression of microRNA-454 in TGF-β1-stimulated hepatic stellate cells and in mouse livers infected with Schistosoma japonicum.

Zhu D, He X, Duan Y, Chen J, Wang J, Sun X, Qian H, Feng J, Sun W, Xu F, Zhang L - Parasit Vectors (2014)

Bottom Line: The results showed that the expression of miR-454 was down-regulated in the TGF-β1-treated LX-2 cells and miR-454 could inhibit the activation of HSCs by directly targeting Smad4.However, we found that miR-454 had no effect on cell cycle and cell proliferation in TGF-β1-treated LX-2.Besides these, miR-454 was found to be regulated in the process of Schistosoma japonicum infection.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathogen Biology, School of Medicine, Nantong University, 19 Qixiu Road, Nantong, Jiangsu 226001, People's Republic of China. yinongduan@gmail.com.

ABSTRACT

Background: In the process of hepatic fibrosis, hepatic stellate cells (HSCs) can be activated by many inflammatory cytokines. The transforming growth factor-β1 (TGF-β1) is one of the main profibrogenic mediators. Recently, some studies have also shown that microRNAs (miRNAs) play essential roles in the progress of liver fibrosis by being involved in the differentiation, fat metabolism and ECM production of HSCs.

Methods: The expression of miR-454 in LX-2 cells treated with TGF-β1 and in the fibrotic livers with Schistosoma japonicum infection was detected by qRT-PCR. The role of miR-454 on LX-2 cells was then analyzed by Western blot, flow cytometry and luciferase assay.

Results: The results showed that the expression of miR-454 was down-regulated in the TGF-β1-treated LX-2 cells and miR-454 could inhibit the activation of HSCs by directly targeting Smad4. However, we found that miR-454 had no effect on cell cycle and cell proliferation in TGF-β1-treated LX-2. Besides these, miR-454 was found to be regulated in the process of Schistosoma japonicum infection.

Conclusions: All the results suggested that miR-454 could provide a novel therapeutic approach for treating liver fibrosis, especially the liver fibrosis induced by Schistosoma japonicum.

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Related in: MedlinePlus

The effect of miR-454 on proliferation and cell cycle distribution of TGF-β1-treated LX-2. (A) The expression of miR-454 in LX-2 cells was analyzed by qRT-PCR. The expression of miR-454 could be down-regulated in the cells treated with TGF-β1 and in the cells co-treated with TGF-β1 and NS-miRNA. * P <0.05 vs control (cells with no stimulus). However, the down-regulation of miR-454 expression induced by TGF-β1 could be reversed in the cells transfected with miR-454 mimics (# P <0.05, compared with the group of the cells co-treated with TGF-β1 and miR-454 mimics). (B) The proliferation capacity of LX-2 cells was increased when the cells were treated with TGF-β1. * P <0.05 vs control (cells with no stimulus). However, miR-454 mimics could not regulate the proliferation of LX-2 cells by MTT assay (P>0.05, compared with the group of the cells co-treated with TGF-β1 and NS-miRNA). (C) The effect of miR-454 on cell cycle of LX-2 cells was analyzed by flow cytometry. P>0.05 vs control (cells with no stimulus).
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Figure 3: The effect of miR-454 on proliferation and cell cycle distribution of TGF-β1-treated LX-2. (A) The expression of miR-454 in LX-2 cells was analyzed by qRT-PCR. The expression of miR-454 could be down-regulated in the cells treated with TGF-β1 and in the cells co-treated with TGF-β1 and NS-miRNA. * P <0.05 vs control (cells with no stimulus). However, the down-regulation of miR-454 expression induced by TGF-β1 could be reversed in the cells transfected with miR-454 mimics (# P <0.05, compared with the group of the cells co-treated with TGF-β1 and miR-454 mimics). (B) The proliferation capacity of LX-2 cells was increased when the cells were treated with TGF-β1. * P <0.05 vs control (cells with no stimulus). However, miR-454 mimics could not regulate the proliferation of LX-2 cells by MTT assay (P>0.05, compared with the group of the cells co-treated with TGF-β1 and NS-miRNA). (C) The effect of miR-454 on cell cycle of LX-2 cells was analyzed by flow cytometry. P>0.05 vs control (cells with no stimulus).

Mentions: To investigate whether miR-454 was involved in the proliferation of HSCs, miR-454 mimics was transiently transfected into LX-2 cells and the results showed that the miR-454 expression was obviously increased in the cells with TGF-β1 and miR-454 mimics transfection, compared with the cells transfected with NS-mimics and TGF-β1 (Figure 3A). The down-regulation of miR-454 expression induced by TGF-β1 could be reversed in the cells transfected with miR-454 mimics, but not in the cells transfected with NS-miRNA (Figure 3A). Then, we analyzed whether the expression of miR-454 could affect cellular proliferation of LX-2 by MTT assay. The results showed that miR-454 had no effect on the proliferation of LX-2 with the existence of TGF-β1, compared with the group of the cells treated with TGF-β1 and NS-miRNA (P>0.05, Figure 3B). To further observe whether miR-454 could affect cell cycle in TGF-β1-treated LX-2 cells, we analyzed the effect of miR-454 on cell cycle of LX-2 cells by flow cytometry. We found that miR-454 mimics had no significant effect on cell cycle distribution, compared with those cells treated with TGF-β1 and NS-miRNA (P>0.05, Figure 3C). These data indicated that over-expression of miR-454 could not affect the proliferation and cell cycle distribution of TGF-β1-treated-LX-2 cells.


Expression of microRNA-454 in TGF-β1-stimulated hepatic stellate cells and in mouse livers infected with Schistosoma japonicum.

Zhu D, He X, Duan Y, Chen J, Wang J, Sun X, Qian H, Feng J, Sun W, Xu F, Zhang L - Parasit Vectors (2014)

The effect of miR-454 on proliferation and cell cycle distribution of TGF-β1-treated LX-2. (A) The expression of miR-454 in LX-2 cells was analyzed by qRT-PCR. The expression of miR-454 could be down-regulated in the cells treated with TGF-β1 and in the cells co-treated with TGF-β1 and NS-miRNA. * P <0.05 vs control (cells with no stimulus). However, the down-regulation of miR-454 expression induced by TGF-β1 could be reversed in the cells transfected with miR-454 mimics (# P <0.05, compared with the group of the cells co-treated with TGF-β1 and miR-454 mimics). (B) The proliferation capacity of LX-2 cells was increased when the cells were treated with TGF-β1. * P <0.05 vs control (cells with no stimulus). However, miR-454 mimics could not regulate the proliferation of LX-2 cells by MTT assay (P>0.05, compared with the group of the cells co-treated with TGF-β1 and NS-miRNA). (C) The effect of miR-454 on cell cycle of LX-2 cells was analyzed by flow cytometry. P>0.05 vs control (cells with no stimulus).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 3: The effect of miR-454 on proliferation and cell cycle distribution of TGF-β1-treated LX-2. (A) The expression of miR-454 in LX-2 cells was analyzed by qRT-PCR. The expression of miR-454 could be down-regulated in the cells treated with TGF-β1 and in the cells co-treated with TGF-β1 and NS-miRNA. * P <0.05 vs control (cells with no stimulus). However, the down-regulation of miR-454 expression induced by TGF-β1 could be reversed in the cells transfected with miR-454 mimics (# P <0.05, compared with the group of the cells co-treated with TGF-β1 and miR-454 mimics). (B) The proliferation capacity of LX-2 cells was increased when the cells were treated with TGF-β1. * P <0.05 vs control (cells with no stimulus). However, miR-454 mimics could not regulate the proliferation of LX-2 cells by MTT assay (P>0.05, compared with the group of the cells co-treated with TGF-β1 and NS-miRNA). (C) The effect of miR-454 on cell cycle of LX-2 cells was analyzed by flow cytometry. P>0.05 vs control (cells with no stimulus).
Mentions: To investigate whether miR-454 was involved in the proliferation of HSCs, miR-454 mimics was transiently transfected into LX-2 cells and the results showed that the miR-454 expression was obviously increased in the cells with TGF-β1 and miR-454 mimics transfection, compared with the cells transfected with NS-mimics and TGF-β1 (Figure 3A). The down-regulation of miR-454 expression induced by TGF-β1 could be reversed in the cells transfected with miR-454 mimics, but not in the cells transfected with NS-miRNA (Figure 3A). Then, we analyzed whether the expression of miR-454 could affect cellular proliferation of LX-2 by MTT assay. The results showed that miR-454 had no effect on the proliferation of LX-2 with the existence of TGF-β1, compared with the group of the cells treated with TGF-β1 and NS-miRNA (P>0.05, Figure 3B). To further observe whether miR-454 could affect cell cycle in TGF-β1-treated LX-2 cells, we analyzed the effect of miR-454 on cell cycle of LX-2 cells by flow cytometry. We found that miR-454 mimics had no significant effect on cell cycle distribution, compared with those cells treated with TGF-β1 and NS-miRNA (P>0.05, Figure 3C). These data indicated that over-expression of miR-454 could not affect the proliferation and cell cycle distribution of TGF-β1-treated-LX-2 cells.

Bottom Line: The results showed that the expression of miR-454 was down-regulated in the TGF-β1-treated LX-2 cells and miR-454 could inhibit the activation of HSCs by directly targeting Smad4.However, we found that miR-454 had no effect on cell cycle and cell proliferation in TGF-β1-treated LX-2.Besides these, miR-454 was found to be regulated in the process of Schistosoma japonicum infection.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathogen Biology, School of Medicine, Nantong University, 19 Qixiu Road, Nantong, Jiangsu 226001, People's Republic of China. yinongduan@gmail.com.

ABSTRACT

Background: In the process of hepatic fibrosis, hepatic stellate cells (HSCs) can be activated by many inflammatory cytokines. The transforming growth factor-β1 (TGF-β1) is one of the main profibrogenic mediators. Recently, some studies have also shown that microRNAs (miRNAs) play essential roles in the progress of liver fibrosis by being involved in the differentiation, fat metabolism and ECM production of HSCs.

Methods: The expression of miR-454 in LX-2 cells treated with TGF-β1 and in the fibrotic livers with Schistosoma japonicum infection was detected by qRT-PCR. The role of miR-454 on LX-2 cells was then analyzed by Western blot, flow cytometry and luciferase assay.

Results: The results showed that the expression of miR-454 was down-regulated in the TGF-β1-treated LX-2 cells and miR-454 could inhibit the activation of HSCs by directly targeting Smad4. However, we found that miR-454 had no effect on cell cycle and cell proliferation in TGF-β1-treated LX-2. Besides these, miR-454 was found to be regulated in the process of Schistosoma japonicum infection.

Conclusions: All the results suggested that miR-454 could provide a novel therapeutic approach for treating liver fibrosis, especially the liver fibrosis induced by Schistosoma japonicum.

Show MeSH
Related in: MedlinePlus