Zhang Pengnan, Liu Yanmei, Feng Youji, Gao Shujun
Obstetrics and Gynecology Hospital of Fudan University, Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Shanghai, PR China.
Obstetrics and Gynecology Hospital of Fudan University, Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Shanghai, PR China The Diagnosis and Treatment Center of Cervical Disease, Obstetrics and Gynecology Hospital of Fudan University, Shanghai, PR China.
Int J Immunopathol Pharmacol. 2016 Sep;29(3):364-75. doi: 10.1177/0394632016641423. Epub 2016 Apr 4.
The aim of this study was to investigate the relationship between HIF-1α and SNAIL gene expression in the epithelial ovarian cancer (EOC) cell line. EOC cells were treated with hypoxia, hypoxia combined with rapamycin, and control. The expression of HIF-1α and E-cad were assessed by reverse transcription-polymerase chain reaction (RT-PCR) and western blotting. The gene expression of SNAIL was studied by RT-PCR and real-time PCR. RNA interference technology was used to determine the relationship between HIF-1α and SNAIL. The present study indicated that the HIF-1α protein was expressed and increased in EOC cell line. SNAIL mRNA was found to increase and E-cad expression decreased with the time of hypoxia prolonged. Hypoxia increased invasion abilities of EOC cell line, but compared with cells exposed to hypoxia, the change of invasive ability of cells with rapamycin had no effect. The expression of HIF-1α protein and SNAIL mRNA could be inhibited gradually by rapamycin. siRNA of HIF-1α could suppress the expression of SNAIL while siRNA of SNAIL had no influence on HIF-1α protein expression. HIF-1α may be the upstream of the SNAIL gene in EOC. Our data suggested that HIF-1α might be an upregulator of the SNAIL gene and HIF-1α-SNAIL-E-cad pathway may play an important role in EOC invasion and metastasis.
本研究旨在探讨上皮性卵巢癌（EOC）细胞系中低氧诱导因子-1α（HIF-1α）与SNAIL基因表达之间的关系。EOC细胞分别接受缺氧处理、缺氧联合雷帕霉素处理以及作为对照。通过逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法评估HIF-1α和E-钙黏蛋白（E-cad）的表达。通过RT-PCR和实时定量PCR研究SNAIL的基因表达。利用RNA干扰技术确定HIF-1α与SNAIL之间的关系。本研究表明，HIF-1α蛋白在EOC细胞系中表达且增加。随着缺氧时间延长，发现SNAIL mRNA增加而E-cad表达降低。缺氧增加了EOC细胞系的侵袭能力，但与缺氧处理的细胞相比，雷帕霉素处理的细胞侵袭能力变化无影响。雷帕霉素可逐渐抑制HIF-1α蛋白和SNAIL mRNA的表达。HIF-1α的小干扰RNA（siRNA）可抑制SNAIL的表达，而SNAIL的siRNA对HIF-1α蛋白表达无影响。在EOC中，HIF-1α可能是SNAIL基因的上游分子。我们的数据表明，HIF-1α可能是SNAIL基因的上调因子，且HIF-1α-SNAIL-E-cad通路可能在EOC侵袭和转移中起重要作用。
