CAJ | 학술논문

目的探讨吲哚胺2,3-双加氧酶(IDO)对人肝癌血管形成能力的影响及其机制.方法转染SMMC-7721细胞建立稳定表达IDO的细胞株,逆转录-聚合酶链反应(RT-PCR)与Westernblot法检测IDO的表达.(1)设未转染组、空质粒转染组、IDO转染组,利用Transwell小室将3组细胞与人脐静脉内皮细胞( HUVEC)共培养,观察HUVEC成管情况,检测各组培养液中血管内皮生长因子(VEGF)的分泌量.(2)将IDO转染组细胞与HUVEC共培养,加入色氨酸(200 mg/L)、1-D-MT(2.5 mmol/L),观察色氨酸、1-D-MT对HUVEC成管及VEGF分泌的影响.结果 (1)IDO转染组、未转染组与空质粒转染组成管数目分别为24.56±0.16、10.38±0.67、11.02 ±0.14;IDO转染组、未转染组与空质粒转染组分泌的VEGF分别为(727.65±0.48)、(512.05±1.11)、(519.35 ±0.38) ng/L;差异均有统计学意义(P＜0.05).(2) IDO转染组、色氨酸组与1-D-MT组成管数目分别为24.84±0.54、15.14±0.24、14.68±0.84; IDO转染组、色氨酸组与1-D-MT组分泌的VEGF分别为(727.65±0.83)、(576.00 ±5.83)、(574.65 ±0.79) ng/L,差异均有统计学意义(P＜0.05).结论 IDO转染的SMMC-7721细胞通过代谢色氨酸途径影响VEGF的表达,进而影响血管形成.
목적 탐토신타알2,3-쌍가양매(IDO)대인간암혈관형성능력적영향급기궤제.방법 전염SMMC-7721세포건립은정표체IDO적세포주,역전록-취합매련반응(RT-PCR)여Westernblot법검측IDO적표체.(1)설미전염조、공질립전염조、IDO전염조,이용Transwell소실장3조세포여인제정맥내피세포( HUVEC)공배양,관찰HUVEC성관정황,검측각조배양액중혈관내피생장인자(VEGF)적분비량.(2)장IDO전염조세포여HUVEC공배양,가입색안산(200 mg/L)、1-D-MT(2.5 mmol/L),관찰색안산、1-D-MT대HUVEC성관급VEGF분비적영향.결과 (1)IDO전염조、미전염조여공질립전염조성관수목분별위24.56±0.16、10.38±0.67、11.02 ±0.14;IDO전염조、미전염조여공질립전염조분비적VEGF분별위(727.65±0.48)、(512.05±1.11)、(519.35 ±0.38) ng/L;차이균유통계학의의(P＜0.05).(2) IDO전염조、색안산조여1-D-MT조성관수목분별위24.84±0.54、15.14±0.24、14.68±0.84; IDO전염조、색안산조여1-D-MT조분비적VEGF분별위(727.65±0.83)、(576.00 ±5.83)、(574.65 ±0.79) ng/L,차이균유통계학의의(P＜0.05).결론 IDO전염적SMMC-7721세포통과대사색안산도경영향VEGF적표체,진이영향혈관형성.
Objective To investigate the effect of indoleamine 2,3-dioxygenase (IDO) on tumor angiogenesis of hepatocellular carcinoma (HCC) cells and the possible mechanism.Methods By using the lipofectamineTM 2000 the eukaryotic expression plasmid pcDNA3.1-IDO and empty vector pcDNA3.1 were transfected into SMMC-7721 cells.After selection with G418,the single cell clone was chosen and cultured.The expression of IDO was detected by using reverse transcription polymerase chain reaction (RT-PCR) and Western blotting.Cells were divided into the non-transfection group,the empty plasmid transfection group,and the IDO transfection group.They were then co-cultured with human umbilical vein endothelial cells (HUVECs) by Transwell.Angiogenesis of HUVECs were observed,and the expression ofvascular endothelial growth factor (VEGF) in every group was detected.The cells in IDO transfection group were co-cultured with HUVECs,1-methyl-D-tryptophan (1-D-MT) (2.5 mmol/L) and tryptophan (200 mg/L) were added separately,and the angiogenesis of HUVECs and the expression of VEGF were observed.Results ( 1 ) The number of neovessels in the IDO transfection group was significantly more than that in the non-transfection group and the empty plasmid transfection group (24.56 ± 0.16,10.38 ± O.67,11.02 ± 0.14 ) ( P ＜ 0.05 ) ; The expression of VEGF in the IDO transfection group was significantly higher than in the non-transfection group and the empty plasmid transfection group (727.65 ±0.48),(512.05 ±1.11 ),(519.35 ±0.38) ng/L (P ＜0.05) ; (2) The number of neovessels in the IDO transfection group was more than that in the tryptophan group and the 1-D-MT group (24.84 ±0.54,15.14 ±0.24,14.68 ±O.84) P ＜ 0.05 ; The expression of VEGF in the IDO transfection group was higher than that in the tryptophan group and the 1 -D-MT group (727.65 ± 0.83 ),( 576.00 ± 5.83 ),(574.65 ± 0.79 ) ng/L ( P ＜0.05).Conclusion Stable transfection of pcDNA3.1-IDO plasmid into SMMC-7721 cells can influence the tumor angiogenesis by regulating the expression of VEGF through tryptophan metabolism passway.