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目的探讨七甲川花菁类荧光小分子IR-780联合阿霉素对肝癌干细胞样细胞的杀伤作用。方法利用CCK-8试剂盒检测不同浓度IR-780(0、1.250、2.500、5.000、10.000、20.000μmol/L)及IR-780联合阿霉素(0、0.025、0.050、0.100、0.200、0.400μmol/L)对4种不同肝癌细胞(QGY-7703、SMMC-7721、Huh-7和Hep G2)的杀伤效应,并确定后续实验IR-780和阿霉素的浓度,采用细胞划痕实验检测IR-780联合阿霉素对肝癌细胞迁移能力的影响;采用无血清悬浮培养的方法从Huh-7细胞株中分离并鉴定出具有干细胞特性的肿瘤细胞,检测IR-780联合阿霉素对肝癌干细胞样细胞的杀伤作用及迁移能力的影响;在裸鼠皮下荷瘤模型中观察IR-780联合阿霉素对肝癌干细胞样细胞生长的抑制作用。结果低浓度的IR-780对肝癌细胞无明显杀伤作用;采用1.250μmol/L的IR-780联合阿霉素可对多种肝癌细胞产生较强的杀伤作用,其中半数致死浓度LC50较单药阿霉素降低25%;采用无血清悬浮培养的方法培养出Huh-7细胞肿瘤球,比较其与贴壁细胞的克隆形成能力、致瘤能力和细胞周期分布,证明悬浮培养的肿瘤细胞具有肿瘤干细胞特性;IR-780联合阿霉素对肝癌干细胞样细胞具有较强的杀伤作用,单用阿霉素和IR-780联合阿霉素对肝癌干细胞样细胞的LC50差异有统计学意义[(0.30±0.02)vs(0.22±0.04)μmol/L,P<0.05];IR-780联合阿霉素可明显抑制肝癌干细胞样细胞的迁移能力和裸鼠皮下肿瘤的生长。结论 IR-780可增强阿霉素的抗肿瘤作用,提高对肝癌干细胞样细胞的杀伤效应。
Objective To investigate the killing effect of heptamethrin fluorescent small molecule IR-780 combined with doxorubicin on hepatoma stem cell-like cells. Methods CCK-8 kit was used to detect the effect of different concentrations of IR-780 (0,1.250,2.500,5.000,10.000,20.000μmol / L) and IR-780 combined with doxorubicin (0,0.025,0.050,0.100,0.200,0.400μmol / L) on four different hepatoma cells (QGY-7703, SMMC-7721, Huh-7 and Hep G2), and to determine the concentration of subsequent experiments IR-780 and doxorubicin. -780 combined with doxorubicin on the migration of hepatocellular carcinoma cells; using serum-free suspension culture method from Huh-7 cell lines were isolated and identified with the stem cell characteristics of tumor cells, IR-780 combined with doxorubicin on liver cancer stem cells Like cell killing effect and migration ability; observe the inhibitory effect of IR-780 combined with doxorubicin on the growth of hepatoma stem cell-like cells in a nude mouse subcutaneous tumor-bearing model. Results Low concentrations of IR-780 had no obvious killing effect on hepatocellular carcinoma cells. Adopting 1.250μmol / L IR-780 combined with doxorubicin could produce a strong killing effect on a variety of hepatocellular carcinoma cells. LC50 of LC50 was higher than that of monotherapy The tumor cells in Huh-7 cells were cultured in serum-free suspension culture. The clonogenic capacity, tumorigenicity and cell cycle distribution of Huh-7 cells were compared with that of adherent cells. The results showed that the suspension-cultured tumor cells had tumor stem cells IR507 combined with doxorubicin had a strong killing effect on hepatoma stem cell-like cells. LC50 of doxorubicin and IR-780 combined with doxorubicin alone had statistical significance ([0.30 ± 0.02) vs (0.22 ± 0.04) μmol / L, P <0.05]. IR-780 combined with doxorubicin significantly inhibited the migration of hepatocellular carcinoma-like stem cells and the growth of subcutaneous tumors in nude mice. Conclusion IR-780 can enhance the anti-tumor effect of doxorubicin and increase the killing effect on hepatoma stem cell-like cells.