精氨酸甲基转移酶抑制剂1通过下调蛋白质精氨酸甲基转移酶5表达抑制肝细胞癌生长
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摘要
目的:探讨精氨酸甲基转移酶抑制剂1(AMI-1)通过抑制蛋白质精氨酸甲基转移酶5(PRMT5)对肝癌细胞系SMMC-7721和BEL-7402的抗肿瘤作用。方法:以含有AMI-1(终浓度0.6,1.2,2.4 mmol·L~(-1))的培养基培养SMMC-7721和BEL-7402细胞后,CCK-8试剂盒检测细胞增殖率;菌落形成实验检测细胞菌落形成情况;裸鼠模型肿瘤形成实验评估体内肿瘤生长;流式细胞术分析细胞周期分布。结果:与人正常肝细胞系LO2相比,人肝癌细胞系SMMC-7721和BEL-7402中RPMT5、真核起始因子4E(eIF4E)、细胞周期蛋白D1(cyclin D1)蛋白表达均显著增加(P<0.05)。与对照组相比,AMI-1处理组细胞增殖率、集落形成数量、肿瘤体积、肿瘤重量均显著降低,G0/G1期DNA含量均显著增加,RPMT5、eIF4E、cyclin D1蛋白表达水平均显著降低(P<0.05)。随着AMI-1浓度的增加,各指标差异均有统计学意义(P<0.05)。结论:AMI-1可能通过抑制PRMT5和eIF4E的表达,引起细胞周期G0/G1期阻滞,从而抑制肝癌细胞系SMMC-7721和BEL-7402体内和体外生长,发挥抗肿瘤形成作用。
Objective: To investigate the antitumor effect of arginine methyltransferase inhibitor 1 (AMI-1) on hepatocellular carcinoma cell lines SMMC-7721 and BEL-7402 by inhibiting protein arginine methyltransferase 5 (PRMT5). Methods: After SMMC-7721 and BEL-7402 cells were cultured in the medium containing AMI-1 with the final concentration of 0.6,1.2 and 2.4 mmol·L~(-1),CCK-8 kit was used to detect the cell proliferation rate,colony formation assay was used to detect the cell colony formation,nude mice model tumor formation assay was used to evaluate tumor growth in vivo,and the cell cycle distribution was analyzed by flow cytometry.Results: Compared with those of human normal hepatocyte line LO2,the expressions of RPMT5,eIF4E and cyclin D1 in SMMC-7721 and BEL-7402 cells were significantly increased (P<0.05). Compared with those in the control group,the cell proliferation rate,colony forming quantity,tumor volume and tumor weight significantly decreased,the content of DNA in G0/G1 phase increased significantly,and the expression levels of RPMT5,eIF4E and cyclin D1 were decreased significantly in the AMI-1 treatment group (P< 0.05).With the concentration increase of AMI-1,the difference of each index was statistically significant (P<0.05).Conclusion: AMI-1 may inhibit the growth of SMMC-7721 and BEL-7402 cell lines by inhibiting the expressions of PRMT5 and eIF4E,which leads to cell cycle arrest in G0/G1 phase resulting in anti-tumor effect.
Objective: To investigate the antitumor effect of arginine methyltransferase inhibitor 1 (AMI-1) on hepatocellular carcinoma cell lines SMMC-7721 and BEL-7402 by inhibiting protein arginine methyltransferase 5 (PRMT5). Methods: After SMMC-7721 and BEL-7402 cells were cultured in the medium containing AMI-1 with the final concentration of 0.6,1.2 and 2.4 mmol·L~(-1),CCK-8 kit was used to detect the cell proliferation rate,colony formation assay was used to detect the cell colony formation,nude mice model tumor formation assay was used to evaluate tumor growth in vivo,and the cell cycle distribution was analyzed by flow cytometry.Results: Compared with those of human normal hepatocyte line LO2,the expressions of RPMT5,eIF4E and cyclin D1 in SMMC-7721 and BEL-7402 cells were significantly increased (P<0.05). Compared with those in the control group,the cell proliferation rate,colony forming quantity,tumor volume and tumor weight significantly decreased,the content of DNA in G0/G1 phase increased significantly,and the expression levels of RPMT5,eIF4E and cyclin D1 were decreased significantly in the AMI-1 treatment group (P< 0.05).With the concentration increase of AMI-1,the difference of each index was statistically significant (P<0.05).Conclusion: AMI-1 may inhibit the growth of SMMC-7721 and BEL-7402 cell lines by inhibiting the expressions of PRMT5 and eIF4E,which leads to cell cycle arrest in G0/G1 phase resulting in anti-tumor effect.
引文
1任利,高建.长链非编码RNA在肝癌中作用机制的研究进展[J].检验医学与临床,2016,13(17):2537-2539
2朱凯,战昊,周俭,等.肝细胞肝癌组织异黏蛋白的表达变化及临床意义[J].中国临床医学,2017,24(3):334-338
3 Zhang B,Dong S,Li Z,et al.Targeting protein arginine methyltransferase 5 inhibits human hepatocellular carcinoma growth via the downregulation of beta-catenin[J].J Transl Med,2015,13(1):349
4 Zhang B,Dong S,Zhu R,et al.Targeting protein arginine methyltransferase 5 inhibits colorectal cancer growth by decreasing arginine methylation of e IF4E and FGFR3[J].Oncotarget,2015,6(26):22799-22811
5 Larsen SC,Sylvestersen KB,Mund A,et al.Proteome-wide analysis of arginine monomethylation reveals widespread occurrence in human cells[J].Sci Signal,2016,9(443):rs9
6曾中秋,马玉玲,裴哲,等.蛋白质精氨酸甲基转移酶抑制剂DB75对人膀胱癌T24细胞的抑制作用[J].应用与环境生物学报,2015,21(1):57-60
7 Greenblatt SM,Liu F,Nimer SD.Arginine methyltransferases in normal and malignant hematopoiesis[J].Exp Hematol,2016,44(6):435-441
8 Yang YZ,Bedford MT.Protein arginine methyltransferases and cancer[J].Nat Rev Cancer,2013,13(1):37-50
9李慧,申徐良,左烨.PRMT5与JAKs/STATs通路的相互作用及其机制的研究进展[J].医学综述,2017,23(9):1696-1701
10 Stopa N,Krebs JE,Shechter D.The PRMT5 arginine methyltransferase:many roles in development,cancer and beyond[J].Cell Mol Life Sci,2015,72(11):2041-2059
11张还添.PRMT5的转录调控及其泛素化-蛋白酶体降解机制研究[D].广州:暨南大学博士学位论文,2015
12张尤历,葛璐,陆瑛,等.PRMT5对肝癌细胞增殖及凋亡的影响[J].中国细胞生物学学报,2015,37(12):1639-1645
13 Li T,Kong ANT,Ma Z,et al.Protein arginine methyltransferase 1may be involved in pregnane x receptor-activated overexpression of multidrug resistance 1 gene during acquired multidrug resistant:[J].Oncotarget,2016,7(15):20236-20248
14 Chan-Penebre E,Kuplast KG,Majer CR,et al.A selective inhibitor of PRMT5 with in vivo and in vitro potency in MCL models[J].Nat Chem Biol,2015,11(6):432-437
15 Zhang T,Günther S,Looso M,et al.Prmt5 is a regulator of muscle stem cell expansion in adult mice[J].Nat Commun,2015,6(8):7140-7151
16吴晓斌,许红英,徐慧.miR-203a-3p通过靶向PRMT5抑制胃癌细胞增殖[J].现代肿瘤医学,2017,25(14):2226-2230
17 Attarschneider O,Drucker L,Gottfried M.Migration and epithelialto-mesenchymal transition of lung cancer can be targeted via translation initiation factors e IF4E and e IF4GI[J].Lab Invest,2016,96(9):1004-1015
18 Koh CM,Bezzi M,Guccione E.The Where and the How of PRMT5[J].Current Molecular Biology Reports,2015,1(1):19-28
19 Lim JH,Lee YM,Lee G,et al.PRMT5 is essential for the e IF4E-mediated 5'-cap dependent translation[J].Biochem Biophys Res Commun,2014,452(4):1016-1021
20 Koh CM,Bezzi M,Guccione E.The Where and the How of PRMT5[J].Current Molecular Biology Reports,2015,1(1):19-28
2朱凯,战昊,周俭,等.肝细胞肝癌组织异黏蛋白的表达变化及临床意义[J].中国临床医学,2017,24(3):334-338
3 Zhang B,Dong S,Li Z,et al.Targeting protein arginine methyltransferase 5 inhibits human hepatocellular carcinoma growth via the downregulation of beta-catenin[J].J Transl Med,2015,13(1):349
4 Zhang B,Dong S,Zhu R,et al.Targeting protein arginine methyltransferase 5 inhibits colorectal cancer growth by decreasing arginine methylation of e IF4E and FGFR3[J].Oncotarget,2015,6(26):22799-22811
5 Larsen SC,Sylvestersen KB,Mund A,et al.Proteome-wide analysis of arginine monomethylation reveals widespread occurrence in human cells[J].Sci Signal,2016,9(443):rs9
6曾中秋,马玉玲,裴哲,等.蛋白质精氨酸甲基转移酶抑制剂DB75对人膀胱癌T24细胞的抑制作用[J].应用与环境生物学报,2015,21(1):57-60
7 Greenblatt SM,Liu F,Nimer SD.Arginine methyltransferases in normal and malignant hematopoiesis[J].Exp Hematol,2016,44(6):435-441
8 Yang YZ,Bedford MT.Protein arginine methyltransferases and cancer[J].Nat Rev Cancer,2013,13(1):37-50
9李慧,申徐良,左烨.PRMT5与JAKs/STATs通路的相互作用及其机制的研究进展[J].医学综述,2017,23(9):1696-1701
10 Stopa N,Krebs JE,Shechter D.The PRMT5 arginine methyltransferase:many roles in development,cancer and beyond[J].Cell Mol Life Sci,2015,72(11):2041-2059
11张还添.PRMT5的转录调控及其泛素化-蛋白酶体降解机制研究[D].广州:暨南大学博士学位论文,2015
12张尤历,葛璐,陆瑛,等.PRMT5对肝癌细胞增殖及凋亡的影响[J].中国细胞生物学学报,2015,37(12):1639-1645
13 Li T,Kong ANT,Ma Z,et al.Protein arginine methyltransferase 1may be involved in pregnane x receptor-activated overexpression of multidrug resistance 1 gene during acquired multidrug resistant:[J].Oncotarget,2016,7(15):20236-20248
14 Chan-Penebre E,Kuplast KG,Majer CR,et al.A selective inhibitor of PRMT5 with in vivo and in vitro potency in MCL models[J].Nat Chem Biol,2015,11(6):432-437
15 Zhang T,Günther S,Looso M,et al.Prmt5 is a regulator of muscle stem cell expansion in adult mice[J].Nat Commun,2015,6(8):7140-7151
16吴晓斌,许红英,徐慧.miR-203a-3p通过靶向PRMT5抑制胃癌细胞增殖[J].现代肿瘤医学,2017,25(14):2226-2230
17 Attarschneider O,Drucker L,Gottfried M.Migration and epithelialto-mesenchymal transition of lung cancer can be targeted via translation initiation factors e IF4E and e IF4GI[J].Lab Invest,2016,96(9):1004-1015
18 Koh CM,Bezzi M,Guccione E.The Where and the How of PRMT5[J].Current Molecular Biology Reports,2015,1(1):19-28
19 Lim JH,Lee YM,Lee G,et al.PRMT5 is essential for the e IF4E-mediated 5'-cap dependent translation[J].Biochem Biophys Res Commun,2014,452(4):1016-1021
20 Koh CM,Bezzi M,Guccione E.The Where and the How of PRMT5[J].Current Molecular Biology Reports,2015,1(1):19-28