二氢黄腐酚通过调控HK2表达对人卵巢癌顺铂耐药细胞株SKOV3/DDP的耐药性影响  

Effect of dihydroflavonol on drug resistance of cisplatin-resistant human ovarian cancer cell line SKOV3/DDP by regulating hexokinase 2 expression and its mechanism

作　　者：王慧[1] 王静[1] 闫冬娟 茹晓楠 马春星 王思思[1] 陈江平[1] 康燕华[1] WANG Hui;WANG Jing;YAN Dong-juan;RU Xiao-nan;MA Chun-xing;WANG Si-si;CHEN Jiang-ping;KANG Yan-hua(Department of Obstetrics and Gynecology,The First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,Hebei,China)

机构地区：[1]河北北方学院附属第一医院妇产科,张家口075000

出　　处：《医学研究生学报》2022年第8期806-812,共7页Journal of Medical Postgraduates

基　　金：河北省医学科学研究重点课题计划(20181744);张家口市科学技术局2020年市级科技计划项目(2021101D)。

摘　　要：目的 目前二氢黄腐酚通过调控HK2表达对人卵巢癌顺铂耐药细胞株SKOV3/DDP的耐药性影响及其机制研究较少。文中旨在研究黄腐酚衍生物二氢黄腐酚(DXN)对人卵巢癌顺铂耐药(DDP)细胞株SKOV3/DDP耐药影响和机制。方法 SKOV3/DDP细胞分成对照组、DXN组(4μmol/L DXN)、DDP组(4 mg/L DDP)、DXN+DDP组(4μmol/L DXN+4 mg/L DDP)、DXN+DDP+Vector组(转染空载质粒+4μmol/L DXN+4 mg/L DDP)、DXN+DDP+HK2组(转染HK2过表达载体+4μmol/L DXN+4 mg/L DDP)、si-NC组(转染siRNA control)、si-HK2组(转染HK siRNA)。MTT方法检测其增殖,计算IC50及其逆转倍数,Western blot检测HK2蛋白表达,测定葡萄糖摄取、乳酸产量。结果 DDP单独处理的IC50为16.17mg/L,联合DXN处理的IC50为9.72 mg/L,逆转倍数为1.66。与对照组和DDP组比较,DXN+DDP组的DDPSKOV3/DDP细胞HK2蛋白表达量、葡萄糖相对摄取量、乳酸相对生成量明显降低(P<0.05)。si-NC组、si-HK2组细胞与DDP处理后,IC50分别为17.92、11.17 mg/L,逆转倍数为1.60。与si-NC组比较,si-HK2组SKOV3/DDP细胞HK2蛋白表达量、葡萄糖相对摄取量、乳酸相对生成量明显降低(P<0.05)。与对照组和DXN+DDP+Vector组比较,DXN+DDP+HK2组SKOV3/DDP细胞中HK2蛋白表达量、葡萄糖相对摄取量、乳酸相对生成量明显升高(P<0.05)。计算DXN(0μmol/L)、DXN(4μmol/L)+Vector、DXN(4μmol/L)+HK2细胞与DDP处理后IC50分别为16.59、9.66、13.10 mg/L,DXN对SKOV3/DDP细胞DDP耐药的逆转倍数为1.72,过表达HK2和DXN联合处理对SKOV3/DDP细胞DDP耐药的逆转倍数为1.27。结论 DXN通过抑制HK2进而抑制有氧糖酵解逆转SKOV3/DDP耐药,为研究DXN在卵巢癌耐药中的作用机制奠定了基础。Objective At present, there are few studies on the effect of dihydroxanthohumol on the drug resistance of human ovarian cancer cisplatin-resistant cell line SKOV3/DDP by regulating the expression of hexokinase 2(HK2) and its mechanism. Effect and mechanism of phenol(DXN) on the drug resistance of human ovarian cancer cisplatin-resistant(DDP) cell line SKOV3/DDP. MethodsSKOV3/DDP cells were divided into nine groups: Control, DXN( 4 μmol/L DXN), DDP( 4 mg/L DDP), DXN + DDP( 4 μmol/L DXN, 4 mg/L DDP), DXN + DDP + Vector( transfected with negative control vector, 4 μmol/L DXN, 4 mg/L DDP), DXN + DDP + HK2( transfected with HK2 overexpression vector, 4 μmol/L DXN, 4 mg/L DDP), si-NC( transfected with siRNA control), si-HK2( transfected with HK siRNA). MTT assay was used to detect its proliferation, IC50 and its reversal fold were calculated. Western blot was used to detect HK2 protein expression, glucose uptake and lactic acid production. Results The IC50 of DDP alone was 16.17 mg/L, and the IC50 of DDP combined with DXN was 9.72 mg/L, and the reversal fold was 1.66. Compared with the control group and DDP group, the HK2 protein expression of DDPSKOV3/DDP cells in DXN + DDP group was decreased, and the relative glucose intake and lactic acid production were decreased(P<0.05). After the cells in si-NC group and si-HK2 group were treated with DDP, the IC50 values were 17.92 and 11.17 mg/L, respectively, and the reversal multiple was 1.60. Compared with si-NC group, the expression of HK2 protein, relative glucose intake and relative lactate production in SKOV3/DDP cells in si-HK2 group were decreased(P<0.05). Compared with control group and DXN+DDP+Vector group, HK2 protein expression, relative glucose intake and relative lactate production in SKOV3/DDP cells in DXN + DDP + HK2 group were increased(P<0.05). The IC50 values of DXN(0 μmol/L), DXN(4 μmol/L) + Vector, DXN(4 μmol/L) + HK2 cells treated with DDP were calculated to be 16.59, 9.66, and 13.10 mg/L, respectively. The reversal multiple of DDP resistance of S

关 键 词：二氢黄腐酚 人卵巢癌顺铂耐药细胞株 己糖激酶2 糖酵解 

分 类 号：R737.31[医药卫生—肿瘤]