缺氧诱导因子-1α基因沉默逆转卵巢癌细胞SKOV3/DDP耐药性的机制  

Mechanism of resistance of ovarian cancer cell SKOV3/DDP reversed by HIF-1αgene silencing

作　　者：薛惠荣[1] 王丽萍[2] 耿嘉男[2] 崔满华[3] 

机构地区：[1]中国人民解放军第二二二医院妇产科,吉林吉林132011 [2]吉林大学药学院实验药理与毒理学教研室,吉林长春130021 [3]吉林大学第二医院妇产科,吉林长春130041

出　　处：《吉林大学学报（医学版）》2013年第6期1103-1107,1311,共6页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30973187)

摘　　要：目的:观察缺氧诱导因子-1α(HIF-1α)沉默后人卵巢癌细胞顺铂耐药株SKOV3/DDP的HIF-1α、多药耐药基因1(MDR1)和B细胞淋巴瘤/白血病-2基因(Bcl-2)mRNA及蛋白产物的表达,探讨HIF-1α逆转SKOV3/DDP耐药性的机制。方法:体外培养卵巢癌细胞株SKOV3(敏感组)及其顺铂耐药株SKOV3/DDP(耐药组),部分耐药株转染HIF-1α干扰质粒pshRNA-HIF(转染组)及对照质粒pshRNA-Control(对照组)。RT-PCR法检测各组细胞HIF-1α、MDR1和Bcl-2 mRNA表达量;Western blotting和免疫组织化学法测定HIF-1α、P-gp(MDR1基因编码蛋白)和Bcl-2蛋白的表达量。结果:RT-PCR检测,敏感组和转染组HIF-1α、MDR1和Bcl-2mRNA表达量明显低于耐药组(P<0.05)。Western blotting检测,敏感组和转染组HIF-1α、MDR1和Bcl-2蛋白表达量明显低于耐药组(P<0.05);免疫组织化学法,敏感组和转染组HIF-1α、MDR1和Bcl-2蛋白表达量明显低于耐药组(P<0.05);MDR1、Bcl-2mRNA及蛋白在敏感组与转染组的表达量差异无统计学意义(P>0.05)。HIF-1α表达与MDR1、Bcl-2mRNA表达量均呈正相关关系(r=0.908,P=0;r=0.916,P=0);HIF-1α表达与P-gp、Bcl-2蛋白表达呈正相关关系(r=0.773,P=0.003;r=0.862,P=0)。结论:HIF-1α沉默逆转人卵巢癌细胞顺铂耐药株SOV3/DDP耐药性可能与MDR1和Bcl-2表达降低有关联。Objective To observe the expressions of hypoxia inducible factor-1 alpha （HIF-la）, multidrun resistance （MDR1）, and B cell lymphoma/leukemia-2 （Bcl-2） mRNA and protein in human ovarian cancer DDP resistant strain SKOV3/DDP after HIF-1α silencing, and to explore the mechanism of the resistance of SKOV3/ DDP reversed by HIF-1α gene silencing. Methods The ovarian cancer cell strain SKOV3 （sensitivity group） and DDP resistance strain SKOV3/DDP （resistance group） were cultured in vitro and parts of the SKOV3/DDP were transfected with HIF-1α interfere plasmid pshRNA-HIF （transfection group） or control plasmid pshRNA-Control （control group）. RT-PCR method was used to detect the expression levels of HIF-1α, MDR1 and Bcl-2 mRNA;Western blotting and immunohistochemisty methods were used to determine the expression levels of HIF-1α, P-gp （MDR1 gene encoding protein）, and Bcl-2 protein in the cells. Results The expression levels of HIF-1α, MDR1 and Bcl 2 mRNA in sensitivity group and transfection group were significantly lower than those in resistance group （P〈0.05） detected by RT-PCR; the expression levels of HIF-1α, MDR1, and Bcl-2 protein in sensitivity group and transfection group were significantly lower than those in resistance group （P〈0.05） determined by Western blotting and immunohistochemistry; the expression levels of MDR1, Bcl 2 mRNA and protein in sensitivity group and transfection group had no significant difference （P〈0.05）. The expressions of HIF-1α and MDR1 and Bcl -2 were closely related （correlation coefficient ofmRNA, r=0.908, P=0; r=0.916, P=0; correlation coefficient of protein, r= 0. 773, P=0. 003; r 0. 862, P=0）.Conclusion The resistance of ovarian cancer cell SKOV3 / DDP HIF-1α gene silencing may be related to the down-regulation of MDR1 and Bcl 2 expressions.

关 键 词：卵巢癌 缺氧诱导因子-1Α 多药耐药 B细胞淋巴瘤 白血病一2 

分 类 号：R711.75[医药卫生—妇产科学]