dn HIF-1对宫颈癌细胞生物学行为的影响

Effect of Dominant Negative HIF-1α (dn HIF-1α) on Biological Characteristics of Uterine Cervix Cancer Cells

目的研究抑制性缺氧诱导因子-1α(dn HIF-1α)对人宫颈癌SiHa细胞生物学行为的影响,探讨HIF-1α在SiHa细胞的血管生成、缺氧保护、凋亡、增殖过程中的作用。方法将表达dn HIF-1α的重组质粒pcDNA3.1-dn HIF-1α转染入SiHa细胞,采用免疫细胞化学法和Western Blotting检测dn HIF-1α转染SiHa细胞后其HIF-1α与血管内皮生长因子(VEGF)表达水平的变化;CoCl2化学缺氧法处理细胞,MTT法测定细胞增殖情况,原位缺口末端标记法检测细胞凋亡情况,同时与pcDNA3.1组和未转染组进行比较。结果重组质粒pcDNA3.1-dn HIF-1α转染SiHa细胞后,其HIF-1α蛋白表达水平与其它两组相比差异无统计学意义,而VEGF蛋白表达水平有较明显的降低(P<0.05)。转染dn HIF-1α的细胞,常氧培养和CoCl2诱导的化学缺氧条件下其增殖能力均低于其他两组(P<0.05),CoCl2处理后细胞凋亡增加,与其他两组相比,差异具有统计学意义(P<0.05)。结论dn HIF-1α抑制SiHa细胞的增殖并促进CoCl2化学缺氧引起的细胞凋亡,同时能降低VEGF蛋白的表达。提示dn HIF-1α可望在宫颈癌治疗中发挥作用。

Objective To explore the effect of dominant negative HIF-1α （dn HIF-1α） on biological characteristics of uterine cervix cancer cell SiHa and elucidate the related mechanism. Methods pcDNA3. 1-dn HIF-1α was transfected into SiHa cells. The expression of HIF-1α and VEGF protein were detected by immunocytochemical method and Western Blotting. The growth proliferation of cells was surveyed by the MTT assay and cell apoptosis was detected through TUNEL after treated with COCl2, meanwhile the results were compared with the group transfected with mock plasmid and untransfected group. Resluts After successfully transfected with relevant plasmid, there＇s no obvious difference of expression of HIF-1α among dn HIF-1α group, pcDNA3. 1 group, and untransfected group, however the expression of VEGF of dn HIF-1α group was significantly lower than that of the others （P〈0. 05）. The proliferation ability of dn HIF-1α group was obviously lower than that of the other two （P〈0. 05）, whether it was under normoxia or chemical hypoxia induced by CoCl2. The characteristic apoptotic morphology was most significantly apparent in dn HIF-1α group among these three （P〈0. 05）. Conclusion Domain negative HIF-1α can inhibit the proliferation of uterine cervix cancer cell and accelerate its apoptosis under hypoxia induced by CoCl2, as well as decrease the expression of VEGF protein. The implications of all this were that the domain negative HIF-1α may play an important role in the therapy of uterine cervix cancer.