MTHFD2对肝癌细胞增殖、迁移、侵袭的影响及机制研究  

Effect and mechanism of MTHFD2 on proliferation,migration,and invasion of hepatocellular carcinoma cells

作　　者：高川 何志刚 马敏俊 谢碧云 洪亮 GAO Chuan;HE Zhigang;MA Minjun;XIE Biyun;HONG Liang(Department of Gastroenterology,the First People's Hospital of Fuyang,Hangzhou 311400,China)

机构地区：[1]杭州市富阳区第一人民医院消化内科,311400

出　　处：《国际消化病杂志》2024年第5期324-331,共8页International Journal of Digestive Diseases

摘　　要：目的探究亚甲基四氢叶酸脱氢酶2(MTHFD2)对肝癌细胞增殖、迁移、侵袭能力的影响及作用机制。方法选择2020年4月至2023年8月于杭州市富阳区第一人民医院接受肝癌切除术的58例肝细胞癌(HCC)患者作为研究对象,收集手术切除的HCC组织及癌旁组织,采用免疫组织化学染色法检测组织中MTHFD2表达水平。另选取正常肝细胞系LO2,以及人肝癌细胞系HepG2、Hep3B和SMMC-7721,采用实时荧光定量PCR法和蛋白质印迹法检测各组细胞中MTHFD2表达水平。将HepG2细胞分为空白对照组(仅给予缓冲溶液)、si-NC组(转染si-NC)和si-MTHFD2组(转染si-MTHFD2),比较各组细胞的增殖、迁移和侵袭能力。选取30只BALB/C-nu/nu雄性裸鼠,将转染24 h后处于对数生长期的HepG2细胞种植于裸鼠右侧腋下,每周测算瘤体体积,4周后剥离瘤块称重。结果HCC组织中MTHFD2蛋白阳性表达率为70.69%,显著高于癌旁组织(32.76%),差异具有统计学意义(P<0.05)。与LO2细胞相比,HepG2、Hep3B和SMMC-7721细胞中MTHFD2的蛋白及mRNA表达水平均显著升高(P均<0.05),且HepG2细胞中MTHFD2的蛋白及mRNA表达水平均最高。与空白对照组和si-NC组相比,si-MTHFD2组细胞中MTHFD2的蛋白及mRNA表达水平均显著降低(P均<0.05)。转染72h和96h后,si-MTHFD2组的光密度(OD)450值分别低于空白对照组和si-NC组(P均<0.05)。与空白对照组和si-NC组相比,si-MTHFD2组的细胞划痕愈合率降低,侵袭细胞数目减少,细胞中磷酸肌醇3激酶(PI3K)、磷酸化PI3K(p-PI3K)、蛋白激酶B(AKT)、p-AKT、哺乳动物雷帕霉素靶蛋白(mTOR)、p-mTOR蛋白表达水平均显著降低(P均<0.05)。培养3周后,si-MTHFD2组裸鼠体内肿瘤体积分别小于空白对照组和si-NC组(P均<0.05)。培养4周后称重结果显示,si-MTHFD2组裸鼠体内肿瘤质量分别小于空白对照组和si-NC组(P均<0.05)。结论下调MTHFD2表达可抑制肝癌细胞增殖、迁移、侵袭及体内成瘤能力,其机制可能与负�Objective This paper is to investigate the effects and mechanisms of methylenetetrahydrofolate dehydrogenase 2(MTHFD2)on the proliferation,migration,and invasion of liver cancer cells.Methods A total of 58 patients with hepatocellular carcinoma(HCC)who underwent liver cancer resection at the First People's Hospital of Fuyang of Hangzhou from April 2020 to August 2023 were selected as the study subjects.The surgically resected HCC tissues and adjacent tissues were collected,and the expression level of MTHFD2 in the tissues was detected by immunohistochemical staining.Normal liver cell line LO2,as well as human liver cancer cell lines HepG2,Hep3B,and SMMC-7721 were selected,and real-time fluorescence quantitative PCR and Western blotting were used to detect the expression level of MTHFD2 in each group of cells.The HepG2 cells were divided into the blank control group(given buffer solution only),the si-NC group(transfected with si-NC),and the si-MTHFD2 group(transfected with si-MTHFD2).In addition,the proliferation,migration,and invasion abilities of each group of cells were compared.Thirty BALB/C-nu/nu male nude mice were selected,and the HepG2 cells in logarithmic growth phase after transfection for 24 hours were implanted in the right axilla of the mice.The tumor volume was measured weekly,and after 4 weeks,the tumor was removed and weighed.Results The positive expression rate of MTHFD2 protein in the HCC tissue is 70.69%higher than that in adjacent tissues(32.76%),with a statistically significant difference(P<0.05).Compared with LO2 cells,the protein and mRNA expression levels of MTHFD2 are significantly increased in HepG2,Hep3B,and SMMC-7721 cells(P<0.05),which are the highest in HepG2 cells.Compared with the blank control group and the si-NC group,the protein and mRNA expression levels of MTHFD2 in the si-MTHFD2 group are significantly reduced(P<0.05).After transfection for 72 and 96 hours,the OD450 values of the si-MTHFD2 group are significantly lower than those of the blank control group and the si-NC group(

关 键 词：肝癌细胞 亚甲基四氢叶酸脱氢酶2 增殖 迁移 侵袭 成瘤能力 磷酸肌醇3激酶 蛋白激酶B 哺乳动物雷帕霉素靶蛋白 

分 类 号：R735.7[医药卫生—肿瘤]