基于调控METTL3介导的m^(6)A修饰探讨鳖龙软肝汤抑制HBV原发性肝癌细胞增殖、迁移的作用机理  

Exploration on the mechanism of Bielong Ruangan Decoction inhibiting the proliferation and migration of HBV primary liver cancer cells based on the regulation of METTL3-mediated m6A modification

作　　者：刘彬彬 姜佳呈 周意[1] 王若宇[1] 彭建平[1] 陈斌[1] 孙克伟[1] 伍玉南[1] LIU Bin-bin;JIANG Jia-cheng;ZHOU Yi;WANG Ruo-yu;PENG Jian-ping;CHEN Bin;SUN Ke-wei;WU Yu-nan(First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,Hunan)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《时珍国医国药》2024年第6期1370-1374,共5页Lishizhen Medicine and Materia Medica Research

基　　金：湖南省自然科学基金(2021JJ30526);湖南中医药大学校级科研基金(2020XJJJ0313);湖南中医药大学研究生创新课题(2021CX77)。

摘　　要：目的鳖龙软肝汤是临床中治疗慢性肝炎肝纤维化的重要中药方剂,通过多年临床的实践和诸多的临床研究报道,鳖龙软肝汤临床治疗乙肝相关性肝癌的疗效较确切。该试验通过研究鳖龙软肝汤对METTL3介导m^(6)A修饰的调控,探索其对HBV相关原发性肝癌细胞增殖、迁移的影响。方法将15只SPF级健康的成年SD大鼠分成空白血清组、鳖龙软肝汤含药血清低剂量组、鳖龙软肝汤含药血清中剂量组,根据人与动物体表面积-等效剂量换算方法,空白组按照剂量灌胃生理盐水,鳖龙软肝汤组按照剂量灌胃鳖龙软肝汤,制备含药血清。分别用CCK-8法、细胞划痕试验检测含药血清对HepG2.2.15细胞增殖、迁移能力的影响,用qPCR检测METTL3、SOCS2 mRNA的表达水平,用m^(6)A RNA甲基化试剂盒检测其m^(6)A修饰水平。结果鳖龙软肝汤含药血清能抑制HBV相关原发性肝癌细胞的增殖、迁移,下调HBV相关肝癌细胞中METTL3表达,上调SOCS2的表达水平,降低m^(6)A修饰水平。结论鳖龙软肝汤含药血清对HBV相关原发性肝癌细胞增殖、迁移的抑制作用可能与METTL3介导的m^(6)A修饰水平下降有关,其过程可能与SOCS2的表达上升密切相关。Objective Bielong Ruangan Decoction is an important traditional Chinese medicine prescription in the treatment of chronic hepatitis and liver fibrosis.Through many years of clinical practice and many clinical reports,the therapeutic effect of Bielong Ruangan Decoction on hepatitis B related liver cancer is quite accurate.In this study,we studied the regulation of METTL3 mediated m6A modification by Bielong Ruangan Decoction,explored its effect on the proliferation of HBV-related primary liver cancer cells.Methods 15 healthy adult SD rats were divided into blank serum group,low dose serum of Bielong Ruangan Decoction and medium dose serum of Bielong Ruangan Decoction.According to the conversion method of human and animal body surface area-equivalent dose,the blank group and Bielong Ruangan decoction were given normal saline according to dose,and the Bielong Ruangan decoction group were given Bielong Ruangan decoction according to dose to prepare drug-containing serum.The effect of drug-containing serum on the proliferation and migration of HepG2.2.15 cells was detected by CCK-8 method and cell scratch test,the expression levels of METTL3 and SOCS2 mRNA were detected by qPCR,and the m6A modification level was detected by m6A RNA methylation kit.Results The serum containing Bielong Ruangan Decoction can inhibit the proliferation of HBV-associated primary liver cancer cells,down-regulate the expression of METTL3 in HBV-associated liver cancer cells,up-regulate the expression of SOCS2,and reduce the m6A modification level.Conclusion The inhibitory effect of the drug-containing serum of Bielong Ruangan Decoction on the proliferation of HBV-related primary liver cancer cells may be related to the decrease of m6A modification level mediated by METTL3,which may be closely related to the increase of SOCS2 expression.

关 键 词：鳖龙软肝汤 肝细胞癌 乙型肝炎病毒 METTL3 SOCS2 m^(6)A甲基化 

分 类 号：R256.4[医药卫生—中医内科学]