索拉非尼、奥沙利铂单药及不同联合方案对人肝癌细胞HepG2的抑制作用

Experimental study on hepatocelluar carcinoma cell line HepG2 by sorafenib,oxaliplatin monotherapy or combination therapy

目的探讨索拉非尼、奥沙利铂单药和两药联合应用对人肝癌细胞株Hep G2增殖、迁移和侵袭的影响。方法选择人肝癌细胞株Hep G2并分为6组处理:奥沙利铂(0.5μg/ml)组(O)、索拉非尼(2.0μmol/L)组(S)、同时联合用药(奥沙利铂0.5μg/ml,索拉非尼2.0μmol/L)组(O+S)、奥沙利铂(0.5μg/ml)序贯索拉非尼(2.0μmol/L)组(OS)、索拉非尼(2.0μmol/L)序贯奥沙利铂(0.5μg/ml)组(SO)和加入100μl普通培养液的阴性对照组(C);分别采用MTT法、流式细胞术、免疫荧光技术、迁移与侵袭实验,观察奥沙利铂、索拉非尼单药和两药联合应用对Hep G2细胞增殖、侵袭与转移的作用。结果 MTT法显示,48 h后用药组对Hep G2细胞增殖均有显著的抑制作用,O+S组的细胞抑制率最高,为(72.13±0.99)%,均明显高于O组、S组和SO组(P<0.01),但与OS组的差异无统计学意义(P>0.05);流式细胞术及荧光显微镜观察显示,O+S组处理Hep G2细胞48 h的凋亡率最高,达65.33%,但与OS组的差异无统计学意义(P>0.05);细胞迁移实验表明,O+S组的细胞迁移抑制率为(71.67±6.66)%,均明显高于O组、S组和SO组(P<0.01),但与OS组的差异无统计学意义(P>0.05);O+S组和OS组均表现为两药协同作用(P<0.01),SO组则表现为两药相加作用(P<0.05)。细胞侵袭实验表明,O+S组的细胞侵袭抑制率为(76.00±5.57)%,明显高于O组、S组(P<0.01);O+S组与OS组均表现为两药协同作用。结论同时联用奥沙利铂和索拉非尼或序贯应用两药较其各自单药对人肝癌Hep G2细胞的增殖、迁移、侵袭抑制作用更强,其中先用奥沙利铂序贯索拉非尼与同时联用奥沙利铂和索拉非尼的协同增效作用相似,均优于先用索拉非尼序贯奥沙利铂。

Objective To observe the effect of sorafenib,oxaliplatin alone or in different combination on the proliferation,migration and invasion of human hepatic carcinoma cell line Hep G2. Methods Human hepatic carcinoma cell line Hep G2 was chosen and divided into 6 groups： oxaliplatin group（ 0. 5 μg / ml,O）,sorafenib group（ 2. 0 μmol / L,S）,combination group（ oxaliplatin 0. 5μg / ml,sorafenib 2. 0 μmol /L; O ＋ S）,oxaliplatin sequent by sorafenib group（ oxaliplatin 0. 5 μg /ml,sorafenib 2. 0 μmol /L; OS）,sorafenib sequent by oxaliplatin group（ sorafenib 2. 0 μmol / L,oxaliplatin 0. 5 μg / ml; SO）,and the negative control group（ 100 μl common culture,C）. MTT method,immunofluorescence technique,flow cytometry,migration experiment,and invade experiment were used to observe sorafenib,oxaliplatin alone or in different combination on the proliferation,invasion and metastasis of Hep G2 cells. Results The results determined by MTT method showed that the treatment groups significantly inhibited the proliferation of Hep G2 cells after 48 h. Cell inhibition rate of O ＋ S group was（ 72. 13 ± 0. 99） %,which was significantly higher than that of O,S groups and SO group（ P 0. 01）,except OS group. Flow cytometry and fluorescence microscopy analysis showed that the apoptosis rate of Hep G2 cells in O ＋ S group was the highest（ 65. 33%） at 48 h,but it had no significant difference compared with OS group（ P 0. 05）. Cell migration experiment showed that the inhibition rate of cell migration in O ＋ S group was（ 71. 67 ± 6. 66） %,which was significantly higher than that of O,S groups and SO group（ P 0. 01）,except OS group. O ＋ S group and OS group exhibited synergistic effect of both sorafenib and oxaliplatin（ P 0. 01）,while SO group showed additive effect（ P 0. 05）. Cells invade experiment showed that cell invasion inhibition rate of O ＋ S group was（ 76. 00 ± 5. 57） %,which was significantly higher than that of O,and S groups（ P 0. 01）. There were two drugs synergistic effect in O ＋ S group and OS group. Conclusion Compared with single drug of sorafenib or oxaliplatin,the inhibition of proliferation,migration and invasion of Hep G2 cells treated by combination and sequential application of sorafenib,oxaliplatin are higher. The synergistic effect is similar between oxaliplatin sequent by sorafenib and sorafenib in combination with oxaliplatin,which are better than sorafenib sequent by oxaliplatin.