沉默ADRB2基因对卵巢上皮性癌细胞生物学特性及顺铂敏感性的影响

目的 探讨下调β2型肾上腺素受体(beta-2 adrenergic receptor,ADRB2)基因表达对卵巢上皮性癌细胞顺铂敏感性以及细胞增殖、凋亡的影响及机制。 方法 以人卵巢癌细胞株SKOV3/DDP为研究对象,采用小RNA干扰技术沉默ADRB2的表达,实验分为4组:ADRB2 shRNA1感染组、ADRB2 shRNA2感染组、阴性对照shRNA感染组、未感染的SKOV3/DDP细胞作为空白对照组。采用不同浓度(0、2.5、5、10、20、40 μmol/L)顺铂作用后,采用CCK-8法检测4组卵巢癌细胞的生长抑制率,流式细胞仪检测4组细胞的凋亡率,Western blot法检测4组卵巢癌细胞中caspase-9、Bcl-2及Bax蛋白的表达,采用单因素方差分析进行比较。 结果 不同浓度的顺铂处理后,4组细胞的生长抑制率均逐渐升高,且呈剂量依赖性。2组ADRB2 shRNA感染细胞的顺铂耐药IC50分别为(29±8)μmol/L和(32±11)μmol/L,明显低于阴性对照组[(104±15)μmol/L,P<0.01]和空白对照组[(112±10)μmol/L,P<0.01];4组细胞的凋亡率逐渐增加,且呈剂量依赖性。2组ADRB2 shRNA感染细胞的凋亡率均明显高于阴性对照组(P<0.01)。4组细胞的caspase-9、Bax蛋白的表达水平逐渐增加,且呈浓度依赖性,2组ADRB2 shRNA感染细胞的caspase-9、Bax蛋白水平均明显高于阴性对照组(均P<0.05)。4组细胞的Bcl-2蛋白的表达水平逐渐降低,且呈浓度依赖性,2组ADRB2 shRNA感染细胞的Bcl-2蛋白水平均明显低于阴性对照组(均P<0.05)。 结论 顺铂诱导可使ADRB2基因下调的顺铂耐药卵巢癌细胞的增殖减少、凋亡增加,其可能机制为通过调节caspase-9、Bcl-2和Bax凋亡家族蛋白的表达,从而增加细胞对顺铂的敏感性。 

Effect of ADRB2 silencing on the cisplatin-resistance and related biological behavior of epithelial ovarian cancer

Objective To investigate the effect of ADRB2 silencing on the cisplatin-resistance and the cell proliferation and apoptosis of epithelial ovarian cancer cells. Methods Human epithelial ovarian cancer cell line SKOV3/DDP was selected for the study and small RNA interference technology was used for silencing the expression of ADRB2. The experiment was divided into 4 groups:ADRB2 shRNA1 group, ADRB2 shRNA2 group, negative control group and blank control group. Cells were treated with different concentration of cisplatin (0, 2.5, 5, 10, 20, 40 μmol/L). Cell proliferation was measured by CCK-8 assay, cell apoptosis was measured by flow cytometry, the expression of caspase-9, Bcl-2 and Bax were measured by western blot. Data was analyzed by t test. Results Cell growth inhibitory rate was significantly upregulated in a dose depend manner upon cisplatin treatment in four groups. The IC50 of two ADRB2 shRNA group were (29±8)μmol/L and (32±11)μmol/L, significantly lower than those of negative control group[(104±15)μmol/L, P<0.01] and blank control group[(112±10)μmol/L, P<0.01]. Cell apoptosis rate was upregulated in an dose depend manner upon cisplatin treatment in four groups. Cell apoptosis of two ADRB2 shRNA group were significantly higher than those of negative control group and blank control group (P<0.01). The caspase-9 and Bax expression were increased upon cisplatin treatment. The levels in two ADRB2 shRNA group were significantly higher than those of negative control group and blank control group (P<0.05). The Bcl-2 expression was decreased upon cisplatin treatment. Its level in two ADRB2 shRNA group were significantly lower than those of negative control group and blank control group (P<0.05). Conclusion ADRB2 silencing significantly increase cisplatin-induced apoptosis and reduce the cell proliferation induced by cisplatin in SKOV3/DDP cells through regulating caspase and Bcl-2 apoptosis signaling pathway.