木犀草素对人卵巢癌SKOV3细胞增殖与凋亡的影响  

Effects of Luteolin on Proliferation and Apoptosis of Human Ovarian Cancer Cell Line SKOV3

作　　者：易均路 侯科名 陈蓉[1] YI Junlu;HOU Keming;CHEN Rong(Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Cure of Metabolic Diseases,College of Traditional Chinese Medicine,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学中医药学院中医药防治代谢性疾病重庆市重点实验室

出　　处：《中药新药与临床药理》2020年第2期125-133,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：重庆市科委基金项目（cstc2016shmszx130057）

摘　　要：目的观察木犀草素(Luteolin)对人卵巢癌SKOV3细胞增殖与凋亡的影响,并通过转录组学探讨其分子机制。方法体外培养人卵巢癌细胞,设立空白对照组、Luteolin组(10、20、40μmol·L^-1),给予药物干预48 h。CCK-8法检测细胞增殖抑制率;EdU实验检测细胞增殖能力;流式细胞术检测细胞周期和细胞凋亡率;Hochest33342染色观察细胞的形态学变化;转录组测序分析空白对照组与40μmol·L^-1Luteolin组干预48 h的差异表达基因(differentially expressed genes,DEGs);根据转录组测序结果,采用实时荧光定量PCR法检测各组细胞中CDKN1A、CDC20、GADD45A、ATF4、DDIT3、HSPA1A、HSPA2、HSPA8、HSPB1 mRNA的表达。结果与空白对照组比较,Luteolin组细胞增殖抑制率均显著升高(P<0.05),呈明显的浓度依赖性;Luteolin组EdU阳性细胞率均显著降低(P<0.05);Luteolin组G0/G1期细胞比例均显著降低(P<0.05),S期细胞比例均显著升高(P<0.05),呈明显的浓度依赖性;Luteolin组细胞凋亡率均显著升高(P<0.05),呈明显的浓度依赖性;Luteolin组Hochest33342染色存在细胞凋亡现象;通过转录组测序分析,筛选获得1476个DEGs,挖掘到包括CDKN1A、CDC20、GADD45A、ATF4、DDIT3、HSPA1A、HSPA2、HSPA8、HSPB1调控细胞增殖与凋亡的DEGs;与空白对照组比较,Luteolin组的CDKN1A、GADD45A、ATF4、DDIT3 mRNA表达水平上调(P<0.05),CDC20、HSPA1A、HSPA2、HSPA8、HSPB1 mRNA表达水平下调(P<0.05)。结论 Luteolin可抑制SKOV3细胞的增殖及诱导其凋亡,其机制可能涉及CDKN1A、CDC20、GADD45A、ATF4、DDIT3、HSPA1A、HSPA2、HSPA8、HSPB1多基因的调控。Objective To observe the effects of Luteolin on proliferation and apoptosis of human ovarian cancer SKOV3 cells,and to explore its molecular mechanism through transcriptomics.Methods Human ovarian cancer cells were cultured in vitro.The cells were divided into blank control group and Luteolin groups(10,20,40 μmol·L^-1),and drug was given for 48 hours.CCK-8 assay was used to detect cell proliferation inhibition rate.EdU was used to detect cell proliferation ability.Flow cytometry was used to detect cell cycle and apoptosis rate.The morphological changes of cells were observed by Hochest33342 staining.Differentially expressed genes(DEGs) between the blank control group and 40 μmol·L^-1 Luteolin groups after intervention for 48 hours were analyzed by transcriptome sequencing.According to the results of transcriptome sequencing,the mRNA expression levels of CDKN1 A,CDC20,GADD45 A,ATF4,DDIT3,HSPA1 A,HSPA2,HSPA8 and HSPB1 in cells were validated by Realtime fluorescent quantitative PCR.Results Compared with the control group,the cell inhibition rates of Luteolin groups were significantly increased(P<0.05),which were in a concentration-dependent manner.The rates of EdUpositive cells of Luteolin groups were significantly decreased(P<0.05).The rates of G0/G1 phase cells of Luteolin groups were significantly decreased(P<0.05),and the proportions of cells in S phase were significantly increased(P<0.05),also with a concentration-dependent manner.Apoptosis rates of Luteolin groups were significantly increased(P<0.05),showing a concentration-dependent manner.Hochest33342 staining showed apoptotic morphology of cells in Luteolin groups.1476 DEGs were obtained by transcriptome sequencing analysis,including CDKN1 A,CDC20,GADD45 A,ATF4,DDIT3,HSPA1 A,HSPA2,HSPA8,HSPB1 that regulate cell proliferation and apoptosis.Compared with the control group,the expression levels of CDKN1 A,GADD45 A,ATF4,DDIT3 mRNA were increased(P<0.05),and the expression levels of CDC20,HSPA1A,HSPA2,HSPA8,HSPB1 mRNA were decreased(P<0.05).Conclusion Luteo

关 键 词：木犀草素 人卵巢癌SKOV3细胞 增殖 凋亡 

分 类 号：R285.5[医药卫生—中药学]