CAJ | 학술논문

目的探讨低氧诱导因子-1α(hypoxia inducible factor-1 alpha,HIF-1α)在低氧状态下,对人鼻咽癌CNE-1细胞黏附和侵袭能力的影响及其分子机制.方法 CoCl2模拟肿瘤低氧微环境.分别用RT-PCR和酶联免疫吸附法(ELISA)检测不同低氧时相HIF-1α、E-钙黏蛋白和趋化因子受体4(chemokine receptor 4,CXCR4)在mRNA及蛋白水平的表达情况.采用RNA干扰技术沉默HIF-1α检测其对E-钙黏蛋白、CXCR4蛋白表达的影响.分别采用黏附实验和Transwell侵袭实验检测HIF-1α沉默对CNE-1细胞黏附及侵袭能力的影响.结果 CNE-1细胞在低氧条件下,HIF-1 αmRNA表达相对恒定,而蛋白随低氧时间延长其表达明显升高(P ＜0.05);E-钙黏蛋白在mRNA及蛋白水平表达均显著降低,而CXCR4均显著升高(P＜0.05).有效沉默HIF-1α可明显上调E-钙黏蛋白的蛋白表达,下调CXCR4表达.CNE-1细胞的黏附率和侵袭穿膜细胞数明显少(P＜0.05).结论低氧促进鼻咽癌CNE-1细胞中HIF-1α蛋白表达水平升高.有效沉默HIF-1α可降低CNE-1细胞的黏附和侵袭能力,其分子机制可能与上调E-钙黏蛋白、下调CXCR4表达有关.
목적 탐토저양유도인자-1α(hypoxia inducible factor-1 alpha,HIF-1α)재저양상태하,대인비인암CNE-1세포점부화침습능력적영향급기분자궤제.방법 CoCl2모의종류저양미배경.분별용RT-PCR화매련면역흡부법(ELISA)검측불동저양시상HIF-1α、E-개점단백화추화인자수체4(chemokine receptor 4,CXCR4)재mRNA급단백수평적표체정황.채용RNA간우기술침묵HIF-1α검측기대E-개점단백、CXCR4단백표체적영향.분별채용점부실험화Transwell침습실험검측HIF-1α침묵대CNE-1세포점부급침습능력적영향.결과 CNE-1세포재저양조건하,HIF-1 αmRNA표체상대항정,이단백수저양시간연장기표체명현승고(P ＜0.05);E-개점단백재mRNA급단백수평표체균현저강저,이CXCR4균현저승고(P＜0.05).유효침묵HIF-1α가명현상조E-개점단백적단백표체,하조CXCR4표체.CNE-1세포적점부솔화침습천막세포수명현소(P＜0.05).결론 저양촉진비인암CNE-1세포중HIF-1α단백표체수평승고.유효침묵HIF-1α가강저CNE-1세포적점부화침습능력,기분자궤제가능여상조E-개점단백、하조CXCR4표체유관.
Objective To investagate the effects of HIF-1α on adhesion and invasion of human nasopharyngeal carcinoma CNE-1 cells under hypoxia and underlying molecular mechanisms.Methods CoCl2 was used to mimic tumor hypoxic microenvironment.mRNA and protein expressions of HIF-1α, E-cadherin and CXCR4 in CNE-1 cells at different hypoxic time phases were detected by RT-PCR and ELISA respectively.The influences of silencing HIF-1α using RNA interference on E-cadherin and CXCR4 expressions were evaluated.Adhesion test Transwell invasion test were used to evaluate the effects of HIF-1α gene silencing on cell adhesion and invasion.Results Under hypoxia, HIF-1α mRNA expression in CNE-1 cells was stable, but its protein expression increased obviously (P ＜ 0.05).Both mRNA and protein expressions of E-cadherin were decreased significantly with prolonged hypoxia, while mRNA and protein expressions of CXCR4 increased significantly (P ＜ 0.05).After silencing HIF-1αgene, expression of E-cadherin protein was up-regulated, but with down-regulated expression of CXCR4 protein, with a decrease significantly in adhesion rate or invasive cell number of CNE-1 cells (P ＜ 0.05).Conclusions Hypoxia can increase HIF-1αprotein expression in nasopharyngeal carcinoma cell line CNE-1.Silencing HIF-1α by RNA interference can reduce inhesion and invasion abilities of CNE-1 cells, which may be mediated by down-regulating E-cadherin expression and up-regulating CXCR4 expression.