阿伐麦布对人卵巢癌SK-OV-3细胞株抑制作用的研究
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摘要
目的观察阿伐麦布(10μM)对人卵巢癌SK-OV-3细胞株增殖能力、侵袭和迁移能力的抑制作用,以及对人卵巢癌SK-OV-3细胞株中ACAT-1蛋白表达的影响,从而反映阿伐麦布对人卵巢癌SK-OV-3细胞株抗肿瘤作用的影响。方法采用阿伐麦布(10μM)作用于人卵巢癌SK-OV-3细胞株,建立SK-OV-3细胞株实验模型(ACAT-1组),该实验模型通过阿伐麦布(10μM)诱导时间的不同分为几个亚实验模型(ACAT-1-24,48,72,96组);通过CCK8实验筛选出阿伐麦布(10μM)作用于SK-OV-3卵巢癌细胞株最佳实验组(72 h),即ACAT-1-72组;通过划痕实验测定对照组(亲代卵巢癌SK-OV-3细胞株)与ACAT-1-72组相同时间内增殖能力以及侵袭转移能力的差异;通过Western blotting实验测定对照组与ACAT-1-72组中ACAT-1蛋白的表达差异;通过免疫荧光实验检测对照组与ACAT-1-72组中ACAT-1蛋白的表达差异。结果①与对照组比较,实验组(ACAT-1组)相同时间内肿瘤细胞的增殖能力减弱,呈时间依赖性(P<0.05),且实验组中的ACAT-1-72组增殖能力减弱最明显,作为后续3组实验的最佳实验组;②与对照组比较,实验组(ACAT-1-72组)相同时间内肿瘤细胞的侵袭和迁移能力减弱(P<0.05);③与对照组比较,实验组(ACAT-1-72组)中ACAT-1蛋白表达下降(P<0.05)。结论阿伐麦布(10μM)对人卵巢癌SK-OV-3细胞株可有效发挥抗肿瘤作用,其主要机制与抑制ACAT-1蛋白的表达有关。
Objective To observe the inhibitory effect of avasimibe on the growth, metastasis invasion and the effect of ACAT-1 expression on human ovarian cancer SK-OV-3 cell line, thus reflecting the effect of avasimibe on the anti-tumor effect of human ovarian cancer SK-OV-3 cell line. Methods Avasimibe(10 μM) was applied to human ovarian cancer SK-OV-3 cell line to establish an experimental model of SK-OV-3 cell line(ACAT-1 group); avasimibe cloth was screened by CCK8 experiment(10 μM) which was applied to SKOV3 ovarian cancer cell line for the best time(72 h), i.e., ACAT-1-72 group; the control group(i.e., parental ovarian cancer SK-OV-3 cell line) and ACAT-1 were determined by scratch test. The proliferative capacity and the ability of invasion and metastasis in 72 groups were the same; the expression of ACAT-1 in the control group and ACAT-1-72 group was determined by Western blot; the control group and ACAT-1-72 group were detected by immunofluorescence assay for the difference in expression of ACAT-1. Results Compared with the control group, the proliferation ability of tumor cells in the experimental group(ACAT-1 group) was weakened in a time-dependent manner(P<0.05), and the proliferative ability of the ACAT-1-72 group in the experimental group was the most obvious. As the best experimental group for the subsequent three sets of experiments; compared with the control group, the experimental group(ACAT-1-72 group) had weaker invasion and migration ability of tumor cells in the same period of time(P<0.05); compared with the control group, the expression of ACAT-1 protein in the experimental group(ACAT-1-72 group) was decreased(P<0.05). Conclusion Avasimibe can effectively promote anti-tumor effect on ovarian cancer SK-OV-3 cell lines, and its main mechanism is related to inhibiting the expression of ACAT-1 protein.
Objective To observe the inhibitory effect of avasimibe on the growth, metastasis invasion and the effect of ACAT-1 expression on human ovarian cancer SK-OV-3 cell line, thus reflecting the effect of avasimibe on the anti-tumor effect of human ovarian cancer SK-OV-3 cell line. Methods Avasimibe(10 μM) was applied to human ovarian cancer SK-OV-3 cell line to establish an experimental model of SK-OV-3 cell line(ACAT-1 group); avasimibe cloth was screened by CCK8 experiment(10 μM) which was applied to SKOV3 ovarian cancer cell line for the best time(72 h), i.e., ACAT-1-72 group; the control group(i.e., parental ovarian cancer SK-OV-3 cell line) and ACAT-1 were determined by scratch test. The proliferative capacity and the ability of invasion and metastasis in 72 groups were the same; the expression of ACAT-1 in the control group and ACAT-1-72 group was determined by Western blot; the control group and ACAT-1-72 group were detected by immunofluorescence assay for the difference in expression of ACAT-1. Results Compared with the control group, the proliferation ability of tumor cells in the experimental group(ACAT-1 group) was weakened in a time-dependent manner(P<0.05), and the proliferative ability of the ACAT-1-72 group in the experimental group was the most obvious. As the best experimental group for the subsequent three sets of experiments; compared with the control group, the experimental group(ACAT-1-72 group) had weaker invasion and migration ability of tumor cells in the same period of time(P<0.05); compared with the control group, the expression of ACAT-1 protein in the experimental group(ACAT-1-72 group) was decreased(P<0.05). Conclusion Avasimibe can effectively promote anti-tumor effect on ovarian cancer SK-OV-3 cell lines, and its main mechanism is related to inhibiting the expression of ACAT-1 protein.
引文
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[4] KREUZINGER C,GAMPERL M,WOLF A,et al.Molecular characterization of 7 new established cell lines from high grade serous ovarian cancer[J].Cancer Lett,2015,362(2):218-228.
[5] CHEN X,SONG Q,XIA L.Synergy of dendritic cell vaccines and avasimibe in treatment of head and neck cancer in mice[J].Med Sci Monit,2017,23(1):4471-4476.
[6] ZHANG L,KIM S B,LUITEL K,et al.Cholesterol depletion by TASIN-1 induces apoptotic cell death through the ER Stress/ROS/JNK signaling in colon cancer cells[J].Mol Cancer Ther,2018,17(5):943-951.
[7] LI M,YANG Y,WEI J,et al.Enhanced chemo-immunotherapy against melanoma by inhibition of cholesterol esterification in CD8 T cells[J].Nanomedicine,2018,14(8):2541-2550.
[8] LEE S S,LI J,TAI J N,et al.Avasimibe encapsulated in human serum albumin blocks cholesterol esterification for selective cancer treatment[J].ACS Nano,2015,9(3):2420-2432.
[9] 张冬雅,郭红军,冯巍,等.MEK/ERK信号通路在上皮性卵巢癌顺铂耐药中的参与作用[J].中华全科医学,2018,16(9):1490-1492.
[10] ZHAO H,JIANG H,LI Z,et al.2-Methoxyestradiol enhances radiosensitivity in radioresistant melanoma MDA-MB-435R cells by regulating glycolysis via HIF-1α/PDK1 axisl[J].Int J Oncol,2017,50(5):1531-1540.
[11] LIN R,ELF S,SHAN C,et al.6-Phosphogluconate dehydrogenase links oxidative PPP,lipogenesis and tumour growth by inhibiting LKB1-AMPK signallingl[J].Nat Cell Biol,2015,17(11):1484-1496.
[12] SHIM S H,SUR S,STEELE R,et al.Disrupting cholesterol esterification by bitter melon suppresses triple-negative breast cancer cell growth[J].Mol Carcinog,2018,57(11):1599-1607.
[13] LI J,QU X,TIAN J,et al.Cholesterol esterification inhibition and gemcitabine synergistically suppress pancreatic ductal adenocarcinoma proliferation[J].PLoS One,2018,13(2):e0193318.
[14] GARCIA-BERMUDEZ J.Drugging ACAT1 for cancer therapy[J].Mol Cell,2016,64(5):856-857.
[15] YANG W,BAI Y,XIONG Y,et al.Potentiating the antitumour response of CD8(+) T cells by modulating cholesterol metabolisml[J].Nature,2016,531(7596):651-655.
[16] MA X,BI E,HUANG C,et al.Cholesterol negatively regulates IL-9-producing CD8 T cell differentiation and antitumor activityl[J].J Exp Med,2018,215(6):1555-1569.
[17] HU L,LI J,CAI H,et al.Avasimibe:A novel hepatitis C virus inhibitor that targets the assembly of infectious viral particles[J].Antiviral Res,2017,148(1):5-14.
[18] 刘秋燕.肿瘤微环境中的免疫细胞亚群代谢:促癌还是抑癌?[J].肿瘤代谢与营养电子杂志,2015,2(3):5-9.