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

作　　者：高浩 赵佼 陈威言 季天民 王群[3] 曹媛[3] GAO Hao;ZHAO Jiao;CHEN Wei-yan;JI Tian-min;WANG Qun;CAO Yuan(Graduate School,Liaoning Universityof TraditionaCl hinese Medicine,Shenyang 110847,Liaoning Province,China;College of Traditional Chinese Medicine,Liaoning Universityof TraditionaCl hinese Medicine,Shenyang 110847,Liaoning Province,China;Key Laboratory of Ministry of Education for Traditional Chinese Medicine Viscera-State Theory and Applications,Liaoning Universityof TraditionaCl hinese Medicine,Shenyang 110847,Liaoning Province,China;Gynecology Department,Cancer Hospital of China MedicalUniversity Liaoning Cancer Hospital&Institute,Shenyang 110042,Liaoning Province,China)

机构地区：[1]辽宁中医药大学研究生学院,辽宁沈阳110847 [2]辽宁中医药大学中医学院,辽宁沈阳110847 [3]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 [4]中国医科大学肿瘤医院/辽宁省肿瘤医院妇科,辽宁沈阳110042

出　　处：《中国临床药理学杂志》2024年第22期3264-3269,共6页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(82074145);辽宁省科技厅基金资助项目(2020-MS-228);辽宁中医药大学中医脏象理论及应用教育部重点实验室开放基金资助项目(zyzx2011);辽宁中医药大学大学生创新创业训练计划基金资助项目(202210162017,S202210162023)。

摘　　要：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300μg·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/1×10^(5) 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,HK2 mRNA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(METT3)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 300μg·mL^(-1)Gyp).The glucose consumption in cells were measured by kit.The expression levels of METT3 mRNA were measured byquantitative real-timepolymerase chainreaction(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 R'T-gPCR were used to detected the mRNA expression of glucose transporter 1(GLU71)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)umol/1 x 10^(5)cells;the relative expression levels of METTL3 mRNA were 0.47+0.01,0.30+0.0l and 1.02+0.0l;the apoptosis rates were(13.52+0.50)%,(14.26+1.30)%and(3.50+0.78)96;the above indexes were statistically diferent belween 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+0.040.32+0.01 and 1.01+0.02;the differences of above indexes were statisically 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.

关 键 词：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 

分 类 号：R28[医药卫生—中药学]