摘要
目的观察吉非替尼联合奥沙利铂对人肝癌细胞株SMMC-7721细胞增殖及凋亡的影响。方法取对数生长期SMMC-7721细胞随机分为对照组、吉非替尼组、10 mg·L^-1奥沙利铂组、20 mg·L^-1奥沙利铂组、40 mg·L^-1奥沙利铂组、吉非替尼+10 mg·L^-1奥沙利铂组、吉非替尼+20 mg·L^-1奥沙利铂组及吉非替尼+40 mg·L^-1奥沙利铂组共8组,依次加二甲基亚砜、2×10^-6 mol·L^-1吉非替尼、10 mg·L^-1奥沙利铂、20 mg·L^-1奥沙利铂、40 mg·L^-1奥沙利铂、2×10^-6 mol·L^-1吉非替尼和10 mg·L^-1奥沙利铂、2×10^-6 mol·L^-1吉非替尼和20 mg·L^-1奥沙利铂、2×10^-6 mol·L^-1吉非替尼和40 mg·L^-1奥沙利铂各100μL,每组设5个复孔。采用四甲基偶氮唑盐比色法测各组细胞给药后24、48、72 h的细胞生长抑制率。另取对数生长期SMMC-7721细胞分为对照组、吉非替尼组、20 mg·L^-1奥沙利铂组、吉非替尼+20 mg·L^-1奥沙利铂组。吉非替尼组细胞加入2×10^-6 mol·L^-1吉非替尼100μL,奥沙利铂组细胞加入20 mg·L^-1奥沙利铂100μL,吉非替尼+20 mg·L^-1奥沙利铂组细胞加入2×10^-6 mol·L^-1吉非替尼100μL和20 mg·L^-1奥沙利铂100μL,对照组细胞加入等体积培养液。采用流式细胞术检测4组细胞给药后48 h的细胞周期和细胞凋亡率。结果吉非替尼组和10 mg·L^-1奥沙利铂组在24 h时细胞抑制率与对照组比较差异无统计学意义(P>0.05);其余各组在各时间点细胞抑制率均高于对照组(P<0.05)。3个联合组各时间点细胞抑制率均高于吉非替尼组(P<0.05),吉非替尼+10 mg·L^-1奥沙利铂组各时间点细胞抑制率均高于10 mg·L^-1奥沙利铂组(P<0.05),吉非替尼+20 mg·L^-1奥沙利铂组各时间点细胞抑制率高于吉非替尼组和20 mg·L^-1奥沙利铂组(P<0.05),吉非替尼+40 mg·L^-1奥沙利铂组各时间点细胞抑制率高于吉非替尼组和40 mg·L^-1奥沙利铂组(P<0.05)。吉非替尼+10 mg·L^-1奥沙利铂组、吉非替尼+20 mg·L^-1奥沙利铂组和吉非替尼+40 mg·L^-1奥沙利铂组各时间点细胞抑制率成逐渐增高趋势,3组之间细胞抑制率两两比较差异均有统计学意义(P<0.05)。24 h时,吉非替尼+10 mg·L^-1奥沙利铂组与40 mg·L^-1奥沙利铂组细胞抑制率比较差异无统计学意义(P>0.05);48、72 h时,吉非替尼+10 mg·L^-1奥沙利铂组细胞抑制率高于40 mg·L^-1奥沙利铂组(P<0.05);吉非替尼+10 mg·L^-1奥沙利铂组在各时间点细胞抑制率均高于20 mg·L^-1奥沙利铂组(P<0.05)。吉非替尼+20 mg·L^-1奥沙利铂组在各时间点细胞抑制率均高于40 mg·L^-1奥沙利铂组(P<0.05)。吉非替尼组、10、20、40 mg·L^-1奥沙利铂组细胞抑制率组内各时间点比较差异无统计学意义(P>0.05)。吉非替尼+10 mg·L^-1奥沙利铂组和吉非替尼+40 mg·L^-1奥沙利铂组48、72 h时细胞抑制率均高于24 h(P<0.05);2组在48 h和72 h时细胞抑制率比较差异均无统计学意义(P>0.05)。吉非替尼+20 mg·L^-1奥沙利铂组随作用时间增加细胞抑制率逐渐增加,各时间点两两比较差异均有统计学意义(P<0.05)。吉非替尼组、20 mg·L^-1奥沙利铂组和吉非替尼+20 mg·L^-1奥沙利铂组细胞的细胞周期分布与对照组比较差异有统计学意义(P<0.05)。20 mg·L^-1奥沙利铂组和吉非替尼+20 mg·L^-1奥沙利铂组处于G 0/G 1期细胞比例低于吉非替尼组(P<0.05),处于S期细胞比例高于吉非替尼组(P<0.05);吉非替尼组和20 mg·L^-1奥沙利铂组处于G 2/M期细胞的比例比较差异无统计学意义(P>0.05);吉非替尼+20 mg·L^-1奥沙利铂组处于G 2/M期细胞的比例高于吉非替尼组和20 mg·L^-1奥沙利铂组(P<0.05)。吉非替尼组、20 mg·L^-1奥沙利铂组和吉非替尼+20 mg·L^-1奥沙利铂组细胞凋亡率均高于对照组(P<0.05);吉非替尼组、20 mg·L^-1奥沙利铂组细胞凋亡率比较差异无统计学意义(P>0.05);吉非替尼+20 mg·L^-1奥沙利铂组细胞凋亡率高于吉非替尼组、20 mg·L^-1奥沙利铂组(P<0.05)。结论吉非替尼和奥沙利铂均可抑制肝癌细胞株SMMC-7721的生长和诱导细胞凋亡,吉非替尼联合奥沙利铂后细胞抑制作用明显增强,二者有协同作用。
Objective To observe the effect of gefitinib combined with oxaliplatin on proliferation and apoptosis of human hepatoma cell line SMMC-7721.Methods SMMC-7721cells in logarithmic phase were divided into control group,gefitinib group,10 mg·L^-1 oxaliplatin group,20 mg·L^-1 oxaliplatin group,40 mg·L^-1 oxaliplatin group,gefitinib+10 mg·L^-1 oxaliplatin group,gefitinib+20 mg·L^-1 oxaliplatin group and gefitinib+40 mg·L^-1 oxaliplatin group;five holes was set up in each group.The cells in above eight groups were added with dimethyl sulfoxide 100μL,2×10^-6 mol·L^-1 gefitinib 100μL,10 mg·L^-1 oxaliplatin 100μL,20 mg·L^-1 oxaliplatin 100μL,40 mg·L^-1 oxaliplatin 100μL,gefitinib and 10 mg·L^-1 oxaliplatin 100μL,gefitinib and 20 mg·L^-1 oxaliplatin 100μL,gefitinib and 40 mg·L^-1 oxaliplatin 100μL in turn.The growth inhibition ratio of cells at 24,48,72 h after administration were detected by methyl thiazdyl tetrazolium colorimetry in each group.In addition,SMMC-7721 cells in logarithmic phase were divided into control group,gefitinib group,20 mg·L^-1 oxaliplatin group and gefitinib+20 mg·L^-1 oxaliplatin group.The cells in gefitinib group were added with 2×10^-6 mol·L^-1 gefitinib 100μL,the cells in oxaliplatin group were added with 20 mg·L^-1 oxaliplatin 100μL,the cells in gefitinib+20 mg·L^-1 oxaliplatin group were added with 2×10^-6 mol·L^-1 gefitinib 100μL and 20 mg·L^-1 oxaliplatin 100μL,the cells in control group were added with equal volume of culture medium.The cell cycle and apoptosis rate at 48 hours after administration in the above four groups were detected by flow cytometry.Results There was no significant difference in cell inhibition rate between gefitinib group,10 mg·L^-1 oxaliplatin group and control group at 24 h(P>0.05);the cell inhibition rates in another groups were higher than those in the control group at each time point(P<0.05).The cell inhibition rates of the three combined groups were higher than those in the gefitinib group at each time point(P<0.05);the cell inhibition rate in gefitinib+10 mg·L^-1 oxaliplatin group was higher than that in the 10 mg·L^-1 oxaliplatin group at each time point(P<0.05);the cell inhibition rate in gefitinib+20 mg·L^-1 oxaliplatin group was higher than that in the gefitinib group and 20 mg·L^-1 oxaliplatin group at each time point(P<0.05);the cell inhibition rate in gefitinib+40 mg·L^-1 oxaliplatin group was higher than that in the gefitinib group and 40 mg·L^-1 oxaliplatin group at each time point(P<0.05);the cell inhibition rates in gefitinib+10 mg·L^-1 oxaliplatin group,gefitinib+20 mg·L^-1 oxaliplatin group and gefitinib+40 mg·L^-1 oxaliplatin group increased gradually at each time point,and there was significant difference among the three groups(P<0.05).There was no significant difference in the cell inhibition rate between gefitinib+10 mg·L^-1 oxaliplatin group and 40 mg·L^-1 oxaliplatin group at 24 h(P>0.05);the cell inhibition rate of gefitinib+10 mg·L^-1 oxaliplatin group was higher than that in the 40 mg·L^-1 oxaliplatin group at 48,72 h(P<0.05);the cell inhibition rate of gefitinib+10 mg·L^-1 oxaliplatin group was higher than that in the 20 mg·L^-1 at each time point(P<0.05).The cell inhibition rate of gefitinib+20 mg·L^-1 oxaliplatin group was higher than that in the 40 mg·L^-1 oxaliplatin group at each time point(P<0.05).There was no significant difference in the cell inhibition rate between each time point in gefitinib group,10,20,40 mg·L^-1 oxaliplatin group(P>0.05).The cell inhibition rates of gefitinib+10 mg·L^-1 oxaliplatin group and gefitinib+40 mg·L^-1 oxaliplatin group at 48,72 h were higher than those at 24 h(P<0.05);there was no significant difference in the cell inhibition rates between 48 h and 72 h in the two groups(P>0.05).The cell inhibition rate in gefitinib+20 mg·L^-1 oxaliplatin group increased with the increase of time,and there was significant difference between each time point(P<0.05).There was significant difference in the cell cycle distribution between gefitinib group,20 mg·L^-1 oxaliplatin group,gefitinib+20 mg·L^-1 oxaliplatin group and control group(P<0.05).The proportion of cells in G 0/G 1 phase in 20 mg·L^-1 oxaliplatin group,gefitinib+20 mg·L^-1 oxaliplatin group was lower than that in gefitinib group(P<0.05),and the proportion of cells in S phase was higher than that in gefitinib group(P<0.05);there was no significant difference in the proportion of cells in G 2/M phase between gefitinib group and 20 mg·L^-1 oxaliplatin group(P>0.05);the proportion of cells in G 2/M phase in gefitinib+20 mg·L^-1 oxaliplatin group was higher than that in gefitinib group and 20 mg·L^-1 oxaliplatin group(P<0.05).The apoptosis rates of gefitinib group,20 mg·L^-1 oxaliplatin group and gefitinib+20 mg·L^-1 oxaliplatin group were higher than those in the control group(P<0.05);there was no significant difference in the apoptosis rate between gefitinib group and 20 mg·L^-1 oxaliplatin group(P>0.05);the apoptosis rate of gefitinib+20 mg·L^-1 oxaliplatin group was higher than that in the gefitinib group and 20 mg·L^-1 oxaliplatin group(P<0.05).Conclusion Both gefitinib and oxaliplatin can inhibit the growth and induce apoptosis of SMMC-7721 cell line.The inhibitory effect of gefitinib combined with oxaliplatin on SMMC-7721 cell line is significantly enhanced,and they have synergistic effect.
作者
张海鹏
鲍启德
周利霞
孙李凌
刘军清
赵越
ZHANG Hai-peng;BAO Qi-de;ZHOU Li-xia;SUN Li-ling;LIU Jun-qing;ZHAO Yue(Department of Oncology,Anyang District Hospital,Anyang 455000,Henan Province,China)
出处
《新乡医学院学报》
CAS
2019年第12期1115-1120,共6页
Journal of Xinxiang Medical University
关键词
吉非替尼
奥沙利铂
肝细胞癌
细胞凋亡
gefitinib
oxaliplatin
hepatocellular carcinoma
apoptosis
