纺锤极体成分25通过磷脂酰肌醇3激酶/蛋白激酶B信号通路调控卵巢上皮癌细胞增殖与凋亡的研究  

Spindle pole body component 25 regulates ovarian epithelial cancer cell proliferation and apoptosis by phosphatidylinositide 3-kinase/protein kinase B signaling

作　　者：周筠[1] 汪晶 洪莉[1] ZHOU Yun;WANG Jing;HONG Li(Department of Gynaecology and Obstetrics,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei Province,China)

机构地区：[1]武汉大学人民医院妇产科,湖北武汉430060

出　　处：《中国临床药理学杂志》2023年第7期1004-1008,共5页The Chinese Journal of Clinical Pharmacology

基　　金：湖北省卫生健康委员会科研基金资助项目(WJ2021M154)。

摘　　要：目的 研究纺锤极体成分25(SPC25)对卵巢上皮癌(OEC)细胞增殖和凋亡的影响及其作用机制。方法 将A2780s细胞分为未处理组、对照shRNA组和sh-SPC25干扰组。未处理组细胞常规培养;对照shRNA组细胞培养液加入Control shRNA慢病毒上清液0.5 mL;sh-SPC25干扰组培养液加入sh-SPC25慢病毒上清液0.5 mL。以蛋白质印迹法检测磷脂酰肌醇/蛋白激酶B(PI3K/AKT)信号通路蛋白,以细胞计数-8(CCK-8)法检测细胞增殖情况,以Caspase-Glo 3/7试剂盒及Annexin V/PI双染流式细胞术检测细胞凋亡情况。为进一步验证SPC25的下游调控通路,用PI3K/AKT信号通路激动药胰岛素样生长因子1(IGF-1)100 ng·mL^(-1)处理sh-SPC25干扰组A2780s细胞,记为sh-SPC25+IGF-1组按上述方法检测细胞活性和凋亡的变化。结果 未处理组、对照shRNA组和sh-SPC25干扰组的细胞增殖率分别为(100.16±0.01)%、(102.40±1.55)%和(53.49±3.65)%,Caspase 3/7活性分别为2.11±0.18、1.96±0.09和5.47±0.45,细胞凋亡率分别为(2.33±0.59)%、(3.80±0.10)%和(44.87±1.29)%,p-AKT蛋白相对表达水平分别为0.99±0.03、1.11±0.02和0.46±0.07。sh-SPC25干扰组的上述指标与未处理组、对照shRNA组比较,差异均有统计学意义(均P<0.05)。对照shRNA组、sh-SPC25干扰组和sh-SPC25+IGF-1组的细胞增殖率分别为(103.02±1.10)%、(52.03±8.55)%和(81.43±2.62)%,Caspase 3/7活性分别为1.96±0.36、5.54±0.49和3.51±0.38,细胞凋亡率分别为(2.67±0.31)%、(47.67±1.17)%和(18.87±0.38)%。sh-SPC25+IGF-1组的上述指标与对照shRNA组、sh-SPC25干扰组比较,差异均有统计学意义(均P<0.05)。结论 沉默SPC25可降低OEC细胞的活性,促进其凋亡,作用机制可能与沉默SPC25导致PI3K/AKT信号通路相关蛋白p-AKT表达降低有关。SPC25可作为OEC诊断和预后预测的潜在生物标志物,可作为OEC的候选治疗靶点。Objective To explore the effect of spindle pole body component 25(SPC25)on proliferation and apoptosis of ovarian epithelial cancer(OEC)and its mechanism.Methods A2780s cells were divided into untreated group,control shRNA group and sh-SPC25 group.Cells in untreated group were cultured routinely;0.5 mL of control shRNA lentivirus supernatant was added to the cell culture medium of the control shRNA group;0. 5 mL sh - SPC25 lentivirus supernatant was added to the culture medium ofsh - SPC25 group. The expression of phosphatidylinositol - 3 kinase /protein kinase B ( PI3K/AKT) signaling pathwayproteins was detected by Western blotting. Cell counting kit - 8 ( CCK - 8) was used to detect cell proliferation.Caspase - Glo 3 /7 kit and Annexin V/PI double staining flow cytometry were used to detect cell apoptosis. To furtherverify the downstream regulatory pathway of SPC25,A2780s cells in sh - SPC25 group were treated with 100 ng·mL^(-1)insulin - like growth factor - 1 ( IGF - 1) , recorded as the sh - SPC25 IGF - 1 group,PI3K/AKT signaling agonist,and the changes of cell activity and apoptosis were detected according to the above methods. Results The proliferationrates of untreated group,control shRNA group and sh - SPC25 group were ( 100. 16 ± 0. 01) %,( 102. 40 ± 1. 55) %,( 53. 49 ±3. 65) %;Caspase 3/7 activities were 2. 11 ± 0. 18,1. 96 ±0. 09,5. 47 ± 0. 45;the apoptosis rates were( 2. 33 ± 0. 59) %,( 3. 80 ± 0. 10 ) %,( 44. 87 ± 1. 29 ) %;the expressions of p - AKT were 0. 99 ± 0. 03,1. 11 ± 0. 02,0. 46 ± 0. 07. The difference between sh - SPC25 group and untreated group,control shRNA group werestatistically significant ( P < 0. 05) . The proliferation rate of the control shRNA group,the sh - SPC25 group and thesh - SPC25 + IGF -1 group were ( 103. 02 ±1. 10) %,( 52. 03 ±8. 55) %,( 81. 43 ± 2. 62) %;Caspase 3 /7 activities were1. 96 ± 0. 36,5. 54 ± 0. 49,3. 51 ± 0. 38;the apoptosis rates were ( 2. 67 ± 0. 31 ) %,( 47. 67 ± 1. 17 ) %,( 18. 87 ± 0. 38) %. There were significant differences betw

关 键 词：纺锤极体成分25 卵巢上皮癌 凋亡 蛋白激酶B 胰岛素样生长因子-1 

分 类 号：R97[医药卫生—药品]