Up-regulation of hypoxia inducible factor-1α by cobalt chloride correlates with proliferation and apoptosis in PC-2 cells

Journal of Experimental & Clinical Cancer Research

Xi-Jing Wang²  Hai-Tao Guan²  Zong-Zheng Ji³  Hua-Feng Kang²  Xiao-Xu Liu²  Kun Yan³  Xiao-Bin Ma²  Jie Gao¹  Zhi-Jun Dai² 

[1] Department of Nephrology, the Second Affiliated Hospital, Medical School of Xi'an Jiaotong University, Xi'an, China;Department of Oncology, the Second Affiliated Hospital, Medical School of Xi'an Jiaotong University, Xi'an, China;Department of General Surgery, the Second Affiliated Hospital, Medical School of Xi'an Jiaotong University, Xi'an, China

关键词: Proliferation;    Apoptosis;    HIF-1α;    Cobalt chloride;    Hypoxia;    Pancreatic carcinoma;   

Others  :  826691
DOI  :  10.1186/1756-9966-31-28

 received in 2012-01-13, accepted in 2012-03-27,  发布年份 2012

【 摘 要 】

Background

The exact mechanism of the effects of hypoxia on the proliferation and apoptosis in carcinoma cells is still conflicting. This study investigated the variation of hypoxia-inducible factor-1α(HIF-1α) expression and the apoptosis effect of hypoxia stimulated by cobalt chloride (CoCl₂) in pancreatic cancer PC-2 cells.

Methods

PC-2 cells were cultured with different concentration (50-200 μmol/L) of CoCl₂ after 24-120 hours to simulate hypoxia in vitro. The proliferation of PC-2 cells was examined by MTT assay. The cellular morphology of PC-2 cells were observed by light inverted microscope and transmission electron microscope(EM). The expression of HIF-1α on mRNA and protein level was measured by semi-quantitive RT-PCR and Western blot analysis. Apoptosis of PC-2 cells were demonstrated by flow cytometry with Annexin V-FITC/PI double staining.

Results

MTT assay showed that the proliferation of PC-2 cells were stimulated in the first 72 h, while after treated over 72 h, a dose- dependent inhibition of cell growth could be observed. By using transmission electron microscope, swollen chondrosomes, accumulated chromatin under the nuclear membrane and apoptosis bodies were observed. Flow cytometer(FCM) analysis showed the apoptosis rate was correlated with the dosage of CoCl₂. RT-PCR and Western blot analysis indicated that hypoxia could up-regulate the expression of HIF-1α on both mRNA and protein levels.

Conclusion

Hypoxic microenvironment stimulated by CoCl₂ could effectively induce apoptosis and influence cell proliferation in PC-2 cells, the mechanism could be related to up-expression of HIF-1α.

【 授权许可】

   
2012 Dai et al; licensee BioMed Central Ltd.

【 图 表 】

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