首页|绞股蓝总苷介导METTL3调控m6A甲基化对卵巢癌细胞糖酵解的影响

绞股蓝总苷介导METTL3调控m6A甲基化对卵巢癌细胞糖酵解的影响

Effects of gypenosides mediated METTL3 regulation of m6A methylation on glycolysis in ovarian cancer cells

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目的 探讨绞股蓝总苷(Gyp)对卵巢癌细胞增殖、凋亡和糖酵解的影响,并探讨其对卵巢癌中甲基转移酶样蛋白3(METTL3)介导的N6-甲基腺苷(m6A)甲基化的影响及作用机制.方法 将SKOV3细胞分为对照组(0.9%NaCl)和低、中、高剂量实验组(100、200和300 μg·mL-1 Gyp).用试剂盒测定细胞中葡萄糖消耗量,用实时荧光定量聚合酶链反应(RT-qPCR)法检测细胞中METTL3 mRNA的表达水平,用流式细胞术检测细胞的凋亡情况.裸鼠皮下注射卵巢癌SKOV3细胞构建卵巢癌模型,待造模成功后,随机分为对照组(0.9%NaCl)和低、中、高剂量实验组(50、100和150 mg·kg-1 Gyp),每组10只.用RT-qPCR法检测移植瘤葡萄糖转运蛋白体1(GLUT1)和己糖激酶2(HK2)mRNA的表达水平.结果 在SKOV3细胞中,中、高剂量实验组和对照组的葡萄糖消耗量分别为(7.29±0.67)、(6.15±0.22)和(15.69±0.70)μmol/1 ×105 cells,METTL3 mRNA 相对表达水平分别为 0.47±0.01、0.30±0.01 和1.02±0.01,凋亡率分别为(13.52±0.50)%、(14.26±1.30)%和(3.50±0.78)%,中、高剂量实验组的上述指标与对照组比较,在统计学上差异均有统计学意义(均P<0.01).在卵巢癌移植瘤裸鼠中,中、高剂量实验组和对照组的GLUT1 mRNA相对表达水平分别为0.52±0.03、0.29±0.01和1.02±0.02,HK2mRNA 相对表达水平分别为 0.54±0.04、0.32±0.01 和1.01±0.02,中、高剂量实验组的上述指标与对照组比较,在统计学上差异均有统计学意义(均P<0.01).结论 绞股蓝总苷通过抑制糖酵解在卵巢癌中发挥抗癌作用,其机制与抑制METTL3介导的m6A甲基化有关.
Objective To explore the effects of gypenosides(Gyp)on proliferation,apoptosis,and glycolysis of ovarian cancer cells,and exploring its impact on methyltransferase-like3(METTL3)mediated N6-methyladenosine(m6A)methylation in ovarian cancer and its mechanism of action.Methods The SKOV3 cells were divided into blank group(0.9%NaCl)and experimental-L,-M,-H groups(100,200 and 300 µg·mL-1 Gyp).The glucose consumption in cells were measured by kit.The expression levels of METTL3 mRNA were measured by quantitative real-time polymerase chain reaction(RT-qPCR).The apoptosis was determined by flow cytometry.Ovarian cancer model was constructed by subcutaneous injection of ovarian cancer SKOV 3 cell)in nude mice.After successful mold formation,10 mice were randomly divided into control group(0.9%NaCl)and experimental-L,-M,-H groups(50,100 and 150 mg·kg-1 Gyp),with 10 mice in each group.The RT-qPCR were used to detected the mRNA expression of glucose transporter 1(GLUT1)and hexokinase 2(HK2)in transplanted tumors.Results In SKOV3 cells,the glucose consumption of experimental-M,experimental-H and blank groups were(7.29±0.67),(6.15±0.22)and(15.69±0.70)μmol/1×105 cells;the relative expression levels of METTL3 mRNA were 0.47±0.01,0.30±0.01 and 1.02±0.01;the apoptosis rates were(13.52±0.50)%,(14.26±1.30)%and(3.50±0.78)%;the above indexes were statistically different between the experimental-M,-H groups and the blank group(all P<0.01).In ovarian cancer transplantation tumor in nude mice,the relative expression levels of GLUT1 mRNA in experimental-M,experimental-H and blank groups were 0.52±0.03,0.29±0.01 and 1.02±0.02;the relative expression levels of HK2 mRNA were 0.54±0.04,0.32±0.01 and 1.01±0.02;the differences of above indexes were statistically significant between the experimental-M,-H groups and the blank group(all P<0.01).Conclusion Gypenosides may exert anticancer effects in ovarian cancer by inhibiting glycolysis,which may be related to the inhibition of METTL3 mediated m6A methylation.

gypenosidesovarian cancerglycolysismethyltransferase 3N6-methyladenosinem methylation

高浩、赵佼、陈威言、季天民、王群、曹媛

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辽宁中医药大学研究生学院,辽宁沈阳 110847

中国医科大学肿瘤医院/辽宁省肿瘤医院妇科,辽宁沈阳 110042

辽宁中医药大学中医学院,辽宁沈阳 110847

辽宁中医药大学中医脏象理论及应用教育部重点实验室,辽宁沈阳 110847

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绞股蓝总苷 卵巢癌 糖酵解 甲基转移酶样蛋白3 N6-甲基腺苷甲基化

2024

中国临床药理学杂志
中国药学会

中国临床药理学杂志

CSTPCD北大核心
影响因子:1.91
ISSN:1001-6821
年,卷(期):2024.40(22)