摘要
目的通过RNA干扰技术沉默人子宫颈癌HeLa细胞低氧诱导因子-1α(HIF-1α)的表达,在细胞及移植瘤水平研究低氧对子宫颈癌HeLa细胞多药耐药的影响。方法①将人子宫颈癌HeLa细胞株(记为H0细胞)、转染pGenesil-1空白质粒的HeLa细胞株(记为H1细胞)及转染HIF-1α-shRNA质粒的HeLa细胞株(记为H2细胞)分别行不同时间低氧培养,应用免疫印迹法分析每组HeLa细胞中HIF-1α及多药耐药相关蛋白P-糖蛋白(P-gp)在蛋白水平的表达情况。②应用免疫组织化学法检测HIF-1α及P-gp在上述3种HeLa细胞裸鼠移植瘤模型组织水平的表达情况。结果①免疫印迹法检测发现H0和H1组低氧24h后HIF-1α、P-gp呈强阳性表达,常氧状态下H0、H1和H2组及低氧24h后H2组未见HIF-1α表达,P-gp呈弱阳性表达。②H0、H1、H2组移植瘤中HIF-1α的平均吸光度值分别为(0.5508±0.0771)、(0.5731±0.0538)、(0.1701±0.0884),P-gp的平均吸光度值分别为(0.6976±0.1140)、(0.6964±0.1072)、(0.2138±0.0351)。结论RNA干扰技术沉默人子宫颈癌HeLa细胞低氧诱导因子HIF-1α,可显著降低人子宫颈癌HeLa细胞中HIF-1α及P-gp表达,且二者有明显的相关性。
Objective To study the effect of hypoxia on drug resistance of cervical carcinoma at the cell and xenograft tumor levels by RNA interference suppressing the expression of hypoxia inducible factor-la（HIF-1a） in human cervical carcinoma HeLa cells. Methods Human cervical carcinoma HeLa cell line（marked as H0 cell）, transfection pGenesil-1 HeLa cell line （marked as H1 cell） and transfeetion HIF-1a-shRNA HeLa cell line（marked as H2 cell） were subjected to anaerobic culture for different durations. The expression of HIF-1a and P-gp proteins in each HeLa cell line was detected by Western blot. The expression of HIF-1a and P-gp in the above 3 HeLa cell lines in nude mice xenograft tumor models was detected by immunohistochemical method（IHC）. Results IHC revealed that there was the strong expression of HIF-1a and P-gp after the H0 and H1 cells were subjected to anaerobic culture for 24 h, but the HIF-1a was not expressed and the P-gp exhibited weak positive expression when the H0, H1 and H2 cells were subjected to normal aerobic culture or the H2 cells to anaerobic culture for 24 h. The average absorbance（A） of HIF-1a in the xenograft tumor of the H0, H1 and H2 cells was （0. 550 8±0. 077 1）, （0. 573 1±0.053 8） and （0. 170 1±0.088 4） respectively, while that of the P-gp was （0. 697 6±0. 114 0）, （0. 696 4±0. 107 2） and （0. 213 8±0. 035 1） respectively. Conclusion By RNA interference suppressing the expression of HIF-1a in human cervical carcinoma HeLa cells, the expression of HIF-1a could be reduced remarkably and there was a significant correlation between HIF-1a and P-gp.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2008年第1期81-84,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
湖北省科技攻关计划资助项目(No2005AA301C22)
关键词
低氧
多药耐药
子宫颈癌
hypoxia
multidrug resistance
cervical carcinoma
