Effect of Hypoxia on Epithelia-mesenchymal Transition of Prostate Cancer PC3 Cell
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摘要:目的
探讨低氧对前列腺癌细胞上皮间质转化的影响。
方法分别在常氧（常氧组）和低氧（低氧组）条件下培养PC3细胞24 h，细胞增殖MTT实验检测低氧对前列腺癌PC3细胞增殖力的影响，Transwell侵袭实验评估低氧对前列腺癌PC3细胞侵袭力的影响，Western blot检测HIF-1α、E-cadherin、N-cadherin和Vimentin的表达水平。另外，在常氧条件下用siRNA抑制HIF-1α表达（干扰组），用Western blot检测E-cadherin、N-cadherin和Vimentin的表达水平变化。
结果与常氧组比较，低氧组 前列腺癌PC3细胞的增殖力和侵袭力增加，HIF-1α表达水平增加（P=0.0004），HIF-1α向核内转位；N-cadherin（P<0.0001）和Vimentin（P<0.0001）的表达水平显著增加，E-cadherin的表达水平显著下降（P<0.0001）。同时，干扰组跟常氧组比较，抑制HIF-1α表达使N-cadherin（P=0.0002）和Vimentin（P=0.0002）的表达水平显著下降，而使E-cadherin的表达水平显著增加（P<0.0001）。
结论低氧可能通过调节HIF-1α表达促进前列腺癌PC3细胞的上皮间质转化。
Abstract:ObjectiveTo investigate the effect of hypoxia on epithelia-mesenchymal transition of prostate cancer cells PC3.
MethodsPC3 cells were cultured under aerobic condition (Normal group) and hypoxia condition (Hypoxia group) for 24 h, respectively. Then, the ability of proliferation was detected by MTT, the ability of invasion was assayed by Transwell, and the protein expressions of HIF-1α, E-cadherin, N-cadherin and Vimentin were assayed by Western blot. Meanwhile, PC3 cells were treated with siRNA to inhibit expression of HIF-1α under aerobic condition and then the protein expressions of HIF-1α E-caldherin, N-cadherin and Vimentin were assayed by Western blot.
ResultsCompared with Normal group, the abilities of cell invasion and proliferation were significantly enhanced by hypoxia; and the protein expressions of HIF-1α(P=0.0004), N-cadherin(P<0.0001) and Vimentin(P<0.0001) were significantly increased, but the protein expression of E-cadherin(P<0.0001) was significantly decreased in Hypoxia group; meanwhile, hypoxia promoted HIF-1α to translocate to nucleus.Furthermore, compared with normal group, inhibition expression of HIF-1α by siRNA significantly decreased the protein expressions of N-cadherin(P=0.0002) and Vimentin(P=0.0002), but significantly increased the protein expression of E-cadherin(P<0.0001). C onclusion Hypoxia might promote epithelia-mesenchymal transition of prostate cancer cells PC3 via regulating the expression of HIF-1α.
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Key words:
- Hypoxia /
- HIF-1α /
- Prostate cancer /
- Epithelia-mesenchymal transition
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图 1 低氧使PC3细胞发生显著的形态改变
Figure 1 Hypoxia induced significantly morphological change of PC3 cells
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图 2 Western blot检测常氧和低氧组细胞中各蛋白表达水平
Figure 2 Expressions of HIF-1α,E-cadherin,N-cadherin and Vimentin tested by Western blot in normal and hypoxia groups
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图 3 免疫荧光检测HIF-1α的位置变化
Figure 3 Change of location of HIF-1α was detected by immunofluorescence
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[1] 韩苏军, 张思维, 陈万青, 等. 中国前列腺癌发病现状和流行趋势分析[J]. 临床肿瘤学杂志, 2013, 18(4): 330-4. http://www.cnki.com.cn/Article/CJFDTOTAL-LCZL201304013.htm Han SJ, Zhang SW, Chen WQ, et al. Analysis of the status and trends of prostate cancer incidence in China[J]. Lin Chuang Zhong Liu Xue Za Zhi, 2013, 18(4): 330-4. http://www.cnki.com.cn/Article/CJFDTOTAL-LCZL201304013.htm
[2] Chan N, Milosevic M, Bristow RG. Tumor hypoxia, DNA repair and prostate cancer proregression: new targets and new therapies [J]. Future Oncol, 2007, 3(3): 329-41. doi: 10.2217/14796694.3.3.329
[3] Abrantes AM, Serra E, Goncalves C, et al. Tumour hypoxia and technetium tracers: in vivo studies[J]. Curr Radiopharm, 2012, 5(2): 99-105. doi: 10.2174/1874471011205020099
[4] Stewart GD, Ross JA, McLaren DB, et al. The relevance of a hypoxic tumour microenvironment in prostate cancer[J]. BJU Int, 2009, 105(1): 8-13.
[5] Freedland SJ. Screening, risk assessment, and the approach to therapy in patients with prostate cancer [J]. Cancer, 2011, 117(6): 1123-35. doi: 10.1002/cncr.25477
[6] Emadi Baygi M, Soheili ZS, Schmitz I, et al. Snail regulates cell survival and inhibits cellular senescence in human metastatic prostate cancer cell lines[J]. Cell Biol Toxicol, 2010, 26(6): 553-67. doi: 10.1007/s10565-010-9163-5
[7] Friedl P, Alexander S. Cancer invasion and the microenvironment: plasticity and reciprocity[J]. Cell, 2011, 147(5): 992-1009. doi: 10.1016/j.cell.2011.11.016
[8] Yip WK, Seow HF. Activation of phosphatidylinositol 3-kinase/Akt signaling by EGF downregulates membranous E-cadherin and β-catenin and enhances invasion in nasopharyngeal carcinoma cells[J]. Cancer Lett, 2012, 318(2): 162-72 doi: 10.1016/j.canlet.2011.12.018
[9] Bristow RG, Hill RP. Hypoxia and metabolism. Hypoxia, DNA repair and genetic instability[J]. Nat Rev Cancer, 2008, 8(3):180-92.
[10] Chan DA, Giaccia AJ. Hypoxia, gene expression, and metastasis[J]. Cancer Metastasis Rev, 2007, 26(2): 333-9. doi: 10.1007/s10555-007-9063-1
[11] Yang MH,Wu MZ,Chiou SH,et al. Direct regulation of TWIST by HIF-1alpha promotes metastasis[J]. Nat Cell Biol, 2008, 10(3): 295-305. doi: 10.1038/ncb1691
[12] Milosevic M,Warde P,Ménard C,et al. Tumor hypoxia predicts biochemical failure following radiotherapy for clinically localized prostate cancer[J]. Clinical Cancer Res, 2012, 18(7): 2108-14. doi: 10.1158/1078-0432.CCR-11-2711
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