溶血磷脂酸下调盐酸阿霉素诱导的卵巢癌SKOV3细胞的凋亡

Lysophosphatidic Acid Down-regulated SKOV3 Cells of Ovarian Cancer in Doxorubicin Hydrochloride-induced Apoptosis

溶血磷脂酸(lysophosphatidic acid,LPA)为一种生物活性脂质介质,与卵巢癌发生和发展密切相关。盐酸阿霉素(doxorubicin hydrochloride,DOX)是卵巢癌临床治疗的一线药物。然而,LPA对DOX治疗卵巢癌的影响及其分子机制目前尚不清楚。本论文旨在探讨LPA对DOX治疗卵巢癌的影响及其分子机制,选用人卵巢癌细胞株SKOV3细胞、OVCAR-3细胞和中国仓鼠卵巢CHO细胞,分别用control、LPA(10μmol/L)、DOX(2μmol/L)和LPA(10μmol/L)+DOX(2μmol/L)处理24 h。光学显微镜和透射电子显微镜下观察SKOV3细胞形态,结果显示,LPA减弱DOX诱导的SKOV3细胞死亡和细胞核内染色质边集状况;RT-qPCR结果显示,LPA处理可下调DOX诱导的SKOV3细胞内胱天蛋白酶3基因表达(P<0.05);Western印迹结果显示,LPA处理能下调DOX诱导的SKOV3细胞、OVCAR-3细胞和CHO细胞内胱天蛋白酶3(caspase-3)及切割胱天蛋白酶3(cleaved caspase-3)水平(P<0.05);流式细胞术Annexin V/PI双染结果显示,LPA下调DOX诱导的SKOV3细胞凋亡水平(P<0.05);DCFH-DA法检测SKOV3细胞内活性氧(reactive oxygen species,ROS)水平,发现LPA下调DOX诱导SKOV3细胞内ROS水平(P<0.05)。本论文初步探讨了在LPA干预下DOX对卵巢癌细胞凋亡的影响,为卵巢癌靶向LPA的药物治疗提供参考。

Lysophosphatidic acid(LPA),a bioactive lipid medium,plays an important role in the development and progression of ovarian cancer.Doxorubicin hydrochloride(DOX)is a first-line drug in the ovarian cancer clinical therapy,while the effect and molecular mechanism of LPA in the ovarian cancer with DOX treatment is still unclear.This study intended to explore the effect and molecular mechanism of LPA in ovarian cancer treated with DOX.SKOV3 and OVCAR-3 cells of human ovarian cancer and Chinese hamster ovary cells were treated with control,LPA(10μmol/L),DOX(2μmol/L)and LPA(10μmol/L)+DOX(2μmol/L)respectively for 24 hours.The morphological changes of SKOV3 cells were observed under optical microscope and transmission electron microscope.Results showed that LPA reduced cell death and the degree of chromatin aggregation in SKOV3 cells treated with DOX;RT-qPCR showed that LPA treatment could down-regulate the mRNA levels of caspase-3 in DOX-treated SKOV3 cells(P<0.05);Western blot showed that LPA treatment could reduce caspase-3 and cleaved caspase-3 levels treated with DOX in SKOV3,OVCAR-3 and CHO cells(P<0.05);Flow cytometry using Annexin V/PI double staining showed that LPA could down-regulate apoptosis in SKOV3 cells treated with DOX(P<0.05);DCFH-DA method was used to detect intracellular levels of reactive oxygen species(ROS)in SKOV3 cells.It was found that LPA reduced the intracellular ROS level treated with DOX(P<0.05).Our preliminarily study showed the effect of LPA in the apoptosis of ovarian cancer treated with DOX,which may provide a reference for the drug therapy of ovarian cancer targeting LPA.