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Hypoxia-inducible MiR-182 promotes angiogenesis by targeting RASA1 in hepatocellular carcinoma.

Du C, Weng X, Hu W, Lv Z, Xiao H, Ding C, Gyabaah OA, Xie H, Zhou L, Wu J, Zheng S - J. Exp. Clin. Cancer Res. (2015)

Bottom Line: We found that miR-182 was upregulated in the hypoxia-based microarray.We then revealed that miR-182 was also significantly increased in the HCC tissues compared to the corresponding normal tissues.In addition, the suppression of RASA1 phenocopied the pro-angiogenesis effects of miR-182.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003, China.

ABSTRACT

Background: Hypoxia is a common feature of solid tumors, including HCC. And hypoxia has been reported to play an important role in HCC progression. However, the potential mechanism of miRNAs in hypoxia mediating HCC progression still remains unclear.

Methods: The HCC cells were cultured in the atmosphere of 1 % oxygen to induce hypoxia. The microRNA microarray was employed to search for the hypoxia-inducible miRNAs. RT-PCR, western blot and immunohistochemistry were used to detect the RNA and protein levels. HUVEC were applied to explore the angiogenesis level.

Results: We found that miR-182 was upregulated in the hypoxia-based microarray. We then revealed that miR-182 was also significantly increased in the HCC tissues compared to the corresponding normal tissues. In vitro capilliary tube formation assays showed that the miR-182 promoted angiogenesis. RASA1 was demonstrated as the direct target of miR-182. In addition, the suppression of RASA1 phenocopied the pro-angiogenesis effects of miR-182. Besides, RASA1 was also decreased in the hypoxia HCC cells while the inhibition of miR-182 partially restored the level of RASA1.

Conclusions: Our data showed that hypoxia regulated the expression of miR-182 and RASA1 to promote HCC angiogenesis.

No MeSH data available.


Related in: MedlinePlus

The expression of miR-182 was enhanced in the hypoxia HCC cells. a The level of hif-1a in SK-HEP-1 under different hypoxia exposure time (0 h, 2 h, 8 h, 24 h). b We applied the normoxia SK-HEP-1 cells and hypoxia SK-HEP-1 cells (8 h) for microRNA microarray. The unsupervised hierarchical clustering analysis showed a significant increase of miR-182 in the hypoxia group. c HCC cells were restored to normoxia after applied to normoxia for 8 h. The relative level of miR-182 was detected by RT-PCR. The experiments were performed in three independent times. (**P < 0.01)
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Fig1: The expression of miR-182 was enhanced in the hypoxia HCC cells. a The level of hif-1a in SK-HEP-1 under different hypoxia exposure time (0 h, 2 h, 8 h, 24 h). b We applied the normoxia SK-HEP-1 cells and hypoxia SK-HEP-1 cells (8 h) for microRNA microarray. The unsupervised hierarchical clustering analysis showed a significant increase of miR-182 in the hypoxia group. c HCC cells were restored to normoxia after applied to normoxia for 8 h. The relative level of miR-182 was detected by RT-PCR. The experiments were performed in three independent times. (**P < 0.01)

Mentions: Firstly, we applied SK-HEP-1 cells under hypoxia conditions for several time points. We then examined the hypoxia marker, hypoxia-inducible factor 1 alpha (Hif-1a), in each group. Western blot results showed that the Hif-1a level was significantly increased in the 2 h and 8 h time points, and decreased in the 24 h time point (Fig. 1a). To search for the hypoxia-inducible miRNAs, we performed microRNA microarray scanning of SK-HEP-1 cells subject to normoxia and hypoxia for 8 h. Among all the miRNAs profiled, we identified that miR-182 was the highly induced microRNA in the hypoxia group by setting the fold change > 2 and p value < 0.01 as the cut-off point (Fig. 1b).Fig. 1


Hypoxia-inducible MiR-182 promotes angiogenesis by targeting RASA1 in hepatocellular carcinoma.

Du C, Weng X, Hu W, Lv Z, Xiao H, Ding C, Gyabaah OA, Xie H, Zhou L, Wu J, Zheng S - J. Exp. Clin. Cancer Res. (2015)

The expression of miR-182 was enhanced in the hypoxia HCC cells. a The level of hif-1a in SK-HEP-1 under different hypoxia exposure time (0 h, 2 h, 8 h, 24 h). b We applied the normoxia SK-HEP-1 cells and hypoxia SK-HEP-1 cells (8 h) for microRNA microarray. The unsupervised hierarchical clustering analysis showed a significant increase of miR-182 in the hypoxia group. c HCC cells were restored to normoxia after applied to normoxia for 8 h. The relative level of miR-182 was detected by RT-PCR. The experiments were performed in three independent times. (**P < 0.01)
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4493986&req=5

Fig1: The expression of miR-182 was enhanced in the hypoxia HCC cells. a The level of hif-1a in SK-HEP-1 under different hypoxia exposure time (0 h, 2 h, 8 h, 24 h). b We applied the normoxia SK-HEP-1 cells and hypoxia SK-HEP-1 cells (8 h) for microRNA microarray. The unsupervised hierarchical clustering analysis showed a significant increase of miR-182 in the hypoxia group. c HCC cells were restored to normoxia after applied to normoxia for 8 h. The relative level of miR-182 was detected by RT-PCR. The experiments were performed in three independent times. (**P < 0.01)
Mentions: Firstly, we applied SK-HEP-1 cells under hypoxia conditions for several time points. We then examined the hypoxia marker, hypoxia-inducible factor 1 alpha (Hif-1a), in each group. Western blot results showed that the Hif-1a level was significantly increased in the 2 h and 8 h time points, and decreased in the 24 h time point (Fig. 1a). To search for the hypoxia-inducible miRNAs, we performed microRNA microarray scanning of SK-HEP-1 cells subject to normoxia and hypoxia for 8 h. Among all the miRNAs profiled, we identified that miR-182 was the highly induced microRNA in the hypoxia group by setting the fold change > 2 and p value < 0.01 as the cut-off point (Fig. 1b).Fig. 1

Bottom Line: We found that miR-182 was upregulated in the hypoxia-based microarray.We then revealed that miR-182 was also significantly increased in the HCC tissues compared to the corresponding normal tissues.In addition, the suppression of RASA1 phenocopied the pro-angiogenesis effects of miR-182.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003, China.

ABSTRACT

Background: Hypoxia is a common feature of solid tumors, including HCC. And hypoxia has been reported to play an important role in HCC progression. However, the potential mechanism of miRNAs in hypoxia mediating HCC progression still remains unclear.

Methods: The HCC cells were cultured in the atmosphere of 1 % oxygen to induce hypoxia. The microRNA microarray was employed to search for the hypoxia-inducible miRNAs. RT-PCR, western blot and immunohistochemistry were used to detect the RNA and protein levels. HUVEC were applied to explore the angiogenesis level.

Results: We found that miR-182 was upregulated in the hypoxia-based microarray. We then revealed that miR-182 was also significantly increased in the HCC tissues compared to the corresponding normal tissues. In vitro capilliary tube formation assays showed that the miR-182 promoted angiogenesis. RASA1 was demonstrated as the direct target of miR-182. In addition, the suppression of RASA1 phenocopied the pro-angiogenesis effects of miR-182. Besides, RASA1 was also decreased in the hypoxia HCC cells while the inhibition of miR-182 partially restored the level of RASA1.

Conclusions: Our data showed that hypoxia regulated the expression of miR-182 and RASA1 to promote HCC angiogenesis.

No MeSH data available.


Related in: MedlinePlus